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Liang, Qing You. "Study on supercritical fluid extraction of aristolochic acids in Aristolochia plants." Thesis, University of Macau, 2007. http://umaclib3.umac.mo/record=b1676801.
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Cheung, Thomas Pak Fai, and tom cheung@rmit edu au. "Risk assessment and determination of aristolochic acids and heavy metals in Chinese herbal medicines." RMIT University. Health Sciences, 2007. http://adt.lib.rmit.edu.au/adt/public/adt-VIT20080414.145522.
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There is community concern about toxic contaminants in Chinese herbal medicines. The two areas of contamination that attract most attentions are the nephrotoxic chemical, aristolochic acids found to be present in some Chinese herbs and resulting in renal failure of over 200 patients in Belgium, and heavy metals such as lead, arsenic, cadmium, mercury and chromium, which can cause systemic, CNS, neurological and developmental pathologies. Currently there is a lack of systematic information about the aristolochic acid content in Aristolochia species and related genera, nor have there been any studies on metal contamination conducted in Australia which can provide some scientific basis for assessment of potential risk of Chinese herbal medicines posed to consumers in Australia. This research aimed at addressing these concerns by firstly carrying out a systematic measurement of the contents of aristolochic acids in some relevant raw herbal medicines (CHM) and proprietary medicines (CPM)- 27 CHM, and 7 CPM, and secondly analysing the contents of five heavy metals in 100 CHM, 50 CPM, and 5 commonly used Chinese medicinal formulae (CMF) in the form of raw herbs, and finally evaluating the potential systemic metal toxicity caused by routine ingestions of Chinese medicines in the common form of encapsulated concentrated powder extracts formulated for the treatment of seasonal allergic rhinitis by means of measuring the metal concentrations in blood collected from 71 patients in a randomised double-blind control clinical trial (RCT). Results showed that four of the 37 CHM and two of the 7 CPM contained the banned toxic aristolochic acids. Some of these contaminated medicines could still be purchased over-the-counter in Victoria. Quantitative screening of metal contamination in CHM found that metal concentrations were much lower in the aqueous solutions than in the acid-digested samples. Almost all CHM, CPM and the 5 CMF contained the five heavy metals. Contrary to popular perception, their metal concentrations in the clinically ingested form were extremely low. Their prescribed ingested contents calculated as percentages of the universally recognised regulatory safety standards, the WHO provisional tolerable weekly intake (PTWI), would produce only small percentages of the PTWI for the metal concerned. This was true even when the metal intakes from any forms of Chinese medicines were added to the normal Australian daily dietary metal exposure. These new approaches of analysing the aqueous extractions, as well as interpretation with reference to the WHO regulatory standards and in combination with the Australian normal daily diet, are more relevant and realistic. The RCT in vivo study demonstrated no significant metal accumulation in the blood of both the real treatment group and the placebo control group, thus, attesting to the encouraging finding of the herbal medicine analysis. In conclusion, there is still much to improve in Australia in terms of enforcing the regulation of banning the sale of Chinese herbal medicines that might contain the highly nephrotoxic aristolochic acids. On the other hand, all forms of Chinese medicines in Victoria are safe, and do not appear to pose significant health concerns in terms of metal contamination.
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Alamin, Abdelgadir. "Apport de la chromatographie de partage centrifuge à l'étude phytochimique de 3 plantes utilisées en médecine traditionnelle soudanaise." Thesis, Tours, 2016. http://www.theses.fr/2016TOUR3812/document.
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Ce travail de thèse est une contribution à l’étude phytochimique par Chromatographie de Partage Centrifuge (CPC), de trois plantes utilisées en médecine traditionnelle au Soudan : Aristolochia bracteolata (plante entière), Ziziphus spina-christi (feuilles) et Hydnora abyssinica (rhizomes). Ce travail a permis de mettre au point trois méthodologies de purification par CPC, applicables au fractionnement des acides aristolochiques, des flavonoïdes ou des proanthocyanidols (PAC). Dans ce contexte, la première partie de ce manuscrit est consacrée aux notions générales portant sur la CPC. La deuxième partie porte sur l’étude d’Aristolochia bracteolata. Cette plante est utilisée en médecine traditionnelle, malgré la présence d'acides aristolochiques qui confèrent une néphrotoxicité élevée. Ce travail a permis de mettre au point une méthode innovante pour l’isolement et la purification, avec un très haut niveau de pureté, des acides aristolochiques I, II et IIIa à partir d’un extrait brut, en une étape par CPC en mode d’échange d’ions forts (SIX-CPC). L’acide aristolochique IIIa n’avait jamais été décrit dans cette plante auparavant. Ces résultats ont fait l’objet d’une publication en 2015 dans Separation and Purification Technology. Dans la troisième partie de cette thèse, la CPC a été appliquée à l’isolement de flavonosides présents dans Z. spina-christi. Nous appuyant sur l’expérience du laboratoire dans l’extraction par CPC des flavonosides du Ginkgo biloba, nous proposons une méthodologie de purification utilisant les systèmes de solvant biphasiques EtOAc/n-BuOH/MeOH/H2O et EtOAc/n-BuOH/H2O à différents ratios en fonction de la polarité des flavonosides. Dans la dernière partie, l’étude phytochimique de Hydnora abyssinica a mis en évidence la présence de PACs, polymères de hauts poids moléculaires de flavanols. La méthodologie de fractionnement CPC, précédée d’un pré-fractionnement sur résine LH-20, a permis l’isolement pour la première fois dans cette plante de la katsumadine et du rhodiolosideThis work was a contribution to the phytochemical study of three Sudanese medicinal plants: Aristolochia bracteolata (Whole plant), Ziziphus spina-christi (Leaves) and Hydnora abyssinica (Rhizomes). The specificity of this research program was to emphasize the application of Centrifugal Partition Chromatography (CPC) for the fractionation of these plants. Three specific CPC methodologies were developed for the purification of either aristolochic acids, flavonoids or proanthocyanidins (PACs). In this context, the first part of this manuscript was devoted to the presentation of the CPC methodology. The second part focused on the fractionation of crude extract of Aristolochia bracteolata. This plant is used in traditional medicine, in spite of the presence of aristolochic acids that confer a high nephrotoxicity. In this work was developed an innovating procedure for the isolation and purification in high purity of aristolochic acids I, II and IIIa, in one step from crude extract, using Strong Ions eXchange CPC (SIX-CPC). These results were published in 2015 in Separation and Purification Technology. In the third part, the flavonosides present in Z. spina-christi were isolated using CPC, either in normal or reverse elution mode, using two phases solvent systems EtOAc/n-BuOH/MeOH/H2O or EtOAc/n-BuOH/H2O with different ratios. In the last part, the phytochemical study of Hydnora abyssinica led to the fractionation of PACs, polymers of high molecular weight of flavanols. The CPC fractionation methodology, preceded by LH-20 resin pre-fractionation, allowed the isolation of katsumadine and rhodioloside
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Zhou, Li, and 周莉. "The molecular mechanisms of aristolochic acid nephropathy." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2009. http://hub.hku.hk/bib/B43224349.
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Zhou, Li. "The molecular mechanisms of aristolochic acid nephropathy." Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B43224349.
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Rodriguez, Isela Iveth Gonzales. "Avaliação da atividade antiofídica de \"Aristolochia sprucei\": Isolamento e caracterização estrutural de composto bioativo." Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/60/60134/tde-01092010-005832/.
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Muitas espécies do gênero Aristolochia (Familia Aristolochiaceae) têm sido usadas na medicina tradicional e folclórica como medicamentos e tônicos, as quais demonstravam atividades farmacológicas de interesse clínica e medica como anti-hemorrágica, anti-parasita, antibacteriano, antifúngico, analgésico, antitumoral entre outras. Visando a obtenção de mais informações sobre essas plantas e na procura por substâncias com efeitos antiofídicos, neste trabalho avaliou-se à ação de extratos aquoso, metanólico e de acetato de etila de folhas e caule contra as ações tóxicas da peçonha de Bothrops asper, ambos procedentes do Panamá e contra o efeito miotóxico da peçonha de Bothrops jararacussu e das miotoxinas BthTX-I (isolada de B. jararacussu) e Mtx-II (isolada de B. asper). O extrato das folhas em acetato de etila apresentou a melhor inibição da atividade fosfolipásica da peçonha de B. asper, demonstrando inibição de 45%, 35% e 33% nas proporções de 1:5, 1:10 e 1:30 (m/m), respectivamente. Enquanto que, o extrato de caule em acetato de etila demonstrou maior eficácia na neutralização da atividade coagulante, e, além disso, inibiu 96%, 92% e 87% do edema, da miotoxicidade e hemorragia induzidas pela peçonha de B. asper, respectivamente. Os percentuais diferenciados na neutralização das ações tóxicas da peçonha de Bothrops asper, revelam diferentes perfis do potencial antiofídico de Aristolochia sprucei. Um dos componentes bioativos foi isolado do extrato de caule desta planta por CLAE, e a caracterização química, por ressonância magnética nuclear, demonstrou ser o ácido aristolóquio que inibiu a atividade miotóxica das peçonhas de B. jararacussu e de B. asper em 80% e 85% e assim como a atividade miotóxica da BthTX-I e Mtx-II em 64% e 60%, respectivamente. A atividade hemolítica indireta da peçonha de B. asper foi inibida em 43% pelo o ácido aristolóquio. A análise dos espectros de dicroísmo circular e os estudos de interação por modelagem molecular sugerem que o ácido aristolóquio forma um complexo com a Mtx-II de B. asper inibindo sua atividade. A ligação do ácido aristoloquio com as miotoxinas (MjTX-1, BthTX-II) modificou a forma e a intensidade dos espectros de dicroísmo circular da miotoxina e induziu alterações na porcentagem dos diversos domínios que constituem a estrutura secundária desta miotoxina. Os resultados obtidos confirmam que os extratos de A. sprucei possuem propriedades antiofídicas e sugerem a necessidade de aprofundar estudos que permitam utilizar com segurança os extratos e o principio ativo isolado como suplementos dos antisoros para aumentar a eficácia na neutralização dos efeitos tóxicos locais da peçonha das serpentes.A lot of species of genus Aristolochia (Familia Aristolochiacheae) have been used in traditional medicine and folk, such as medicaments and tonics, which show pharmacological activities of clinic and medical interest, like antihemorragic, antiparasitic, antibacterial, antifungic, analgesic, antitumoral between others. Expecting to get more information about these plants and in the search by substances with antiophidic effects, in this work was evaluated the action of aqueous, metanolic and ethyl acetate extracts from leaves and stems of Aristolochia sprucei against the toxic action of Bothrops asper venom, both native from Panamá and against the myotoxic effect of Bothrops jararacussu venom and BthTX1 (isolated from B. jararacussu) and Mtx-II (isolated from Bothorps asper). The leaves extracts in ethyl acetate showed the best inhibition registered of PLA2 activity of venom de B. asper showing inhibition of 45 %, 35 % and 33 %, in proportion (m/m) of 1:5, 1:10 and 1:30 respectively. As regards to stem extract in ethyl acetate, it showed high efficacy in neutralization of coagulant activity, besides It inhibited 96 %, 92 % and 87 % of edema, myotoxicity and hemorrhage induced by B. asper venom, respectively. One of bioactives components was isolated from stem extract of this plant by CLAE and the chemical characterization by nuclear magnetic resonance, this showed that the compound is the aristolochic acid. This compound inhibited the myotoxic activity of B. jararacussu and B. asper venom in 80 % and 85 %, so like myotoxic activity of BthTx-I and MTx-II in 64 % and 60 % respectively. The indirect hemolytic activity of B. asper venom was inhibited in 43 % by the aristolochic acid. The analyze of spectrum of circular dichroism and the studies of interaction by molecular modelagem suggest that the aristolochic acid forms a complex 1:1 with the miotoxin inhibiting their activity. The joint of aristolochic acid with the miotoxins (MjTX-1, BthTx-II) changes the way and the intensity of spectra from dichroism circular of miotoxin and It induced alteration in percentage of several domains that constitute a secondary structure from this toxin. The results obtained confirm that the extracts of A. sprucei have antiophidic properties and it suggest the necessity of deepen studies that allow to use with safety the extracts and the isolated active principle, like antiserum supplements to increase the efficacy in the neutralization of local toxics effects of snakes venoms.
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Duquesne, Marilyn. "A translational study of the nephrotoxicity of aristolochic acids by a metabonomic approach in NMR spectroscopy validated by conventional biomarkers." Doctoral thesis, Universite Libre de Bruxelles, 2018. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/268946.
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Utilisation de la métabonomique en spectroscopie RMN pour l'identification de biomarqueurs d'exposition à l'acide Aristolochique. Développement de modèles expérimentaux chez des rats mâles. Analyse d'échantillons urinaires provenant de patients croates potentiellement touchés par la Néphropathie endémique des BalkansDoctorat en Sciences biomédicales et pharmaceutiques (Médecine)
info:eu-repo/semantics/nonPublished
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Srđan, Živojinov. "Razvoj animalnog modela nefrotoksične tubulointersticijalne lezije." Phd thesis, Univerzitet u Novom Sadu, Medicinski fakultet u Novom Sadu, 2016. http://www.cris.uns.ac.rs/record.jsf?recordId=99867&source=NDLTD&language=en.
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U eksperimantalnom postupku disertacije miševi NMRI soja su tretirani infuzom biljke Aristolochia clematitis. Sasušeni listovi, grane i plodovi biljke potopljeni su u ključalu vodu i ostavljeni 3-5 sati da stoje, a potom su profiltrirani kroz filter papir. Pravljen je rastvor biljke/vode od 10g/ 1000ml (1%), 20g/ 1000ml (2%) i 40g/ 1000ml (4%). Različite koncentracije infuza su date miševima da piju u neograničenoj količini u periodu od 7 nedelja. Tako su formirane tri ispitne grupe, prva koja je primala 1% infuz, druga 2% infuz i treća 4% infuz i kontrolna grupa koja je dobijala samo vodu da pije. U svakoj grupi je bilo 20 životinja. Tako je razvijen animalni model hronične toksičnosti. Na kraju eksperimenta je urađena patohistološka analiza bubrega, makroskopski pregled organa i merenje diureze tokom trajanja eksperimenta. Urađena je kompletna analiza urina koja podrazumeva utvrđivanje: boje, izgleda, pH, specifične težine, proteina i sedimenta urina. Analize urina ponavljane su na svakih 7 dana u toku 7 nedelja istraživanja. Na kraju eksperimenta urađena je analiza biohemijskih parametara (glukoza, urea, kreatinin, mokraćna kiselina, ukupni bilirubin, direktni bilirubin, ukupni tj. totalni proteini, natrijum i kalijum) i analiza kompletne krvne slike. Utvrđeno je da je Aristolochia clematitis izrazito nefrotoksična biljka. Utvrđene su patohistološke promene tubula i intersitcijuma NMRI miša, koje su bile najveće u ispitnoj grupi koja je primala najaču dozu. Ustanovljene patohistološke promene su slične opisanim patohistološkim promenama tubulointersticijuma bolesnika obolelih od Balkanske endemske nefropatije. Nije ustanovljeno postojanja karcinoma gornjeg urotrakta. Makroskopskim pregledom prilikom obdukcije eksperimentalnih životinja nisu ustanovljene značajnije promene bubrega. Došlo je prvo do izrazitog porasta diureze u prvoj, odnosno drugoj nedelji praćenja, kod druge i treće eksperimentalne grupe, da bi nakon 7 nedelja istaživanja diureza u svim ispitnim grupama bila manja od kontrolne grupe. Postoji porast ureje na kraju istraživanja, koji je dvostruko veći u trećoj eksperimentalnoj grupi u odnosu na kontrolnu. Postoji izrazit pad mokraćne kiseline na kraju istraživanja kod eksperimentalne grupe 3. Postoji izrazit pad granulocita u leukocitarnoj formuli u svim ispitnim grupama, a najveći je u trećoj ispitnoj grupi. Kako je došlo do pada relativnih vrednosti granulocita, tako je došlo do porasta relativnih vrednosti limfocita u prvoj i drugoj ispitnoj grupi. U trećoj ispitnoj grupi je pad granulocita praćen izrazito velikim povećanjem relativnog broja bazofilnih granulocita. Postoji značajan pad specifične težine urina na kraju istraživanja u drugoj i trećoj eksperimentalnoj grupi. Proteinurija je bila čest nalaz svim eksperimentalnim grupama, dok je bila odsutna ili samo u tragu u kontrolnoj grupi. Na kraju eksperimenta je utvrđen znatni porast broja kristala fosfata u eksperimentalnim grupama. Cilindri su se pojavljivali samo u nalazu urina u trećoj ispitnoj grupi. Najveći broj promena urina je utvrđen u trećoj eksperimentlanoj grupi.In the experimental procedure of dissertation, NMRI strain mice were treated with infusion of plants Aristolochia clematitis. Dried leaves, branches and fruit plants are submerged in boiling water and left to stand for 3-5 hours, and then filtered through filter paper. It was made a solution of the plant / water of 10g / 1000ml (1%), 20g / 1000ml (2%) and 40g / 1000ml (4%). Different concentrations of infusions were given to mice to drink an unlimited amount for a period of 7 weeks. So we formed the three test groups, the first who received 1% infusion, the second received 2% infusion and third received 4% infusion and a control group that received only water to drink. In each group there were 20 animals. Thus, developed an animal model of chronic toxicity. At the end of the experiment was performed histopathological analysis of kidneys, macroscopic examination of organs and measuring urine output during the experiment. We performed a complete analysis of urine, which is the determination of: color, appearance, pH, specific gravity, protein and urine sediment. Urinalysis were repeated every 7 days during the 7 weeks of the study. At the end of the experiment were analyzed for biochemical parameters (glucose, urea, creatinine, uric acid, total bilirubin, direct bilirubin, total proteins, sodium and potassium) and analysis of the complete blood count. It has been found that Aristolochia clematitis is extremely nephrotoxic plant. Identified histopathological changes of tubules and interstitium of NMRI mouse, which were the biggest in the test group receiving biggest dose. Established histopathological changes are similar to those described by pathological changes of tubulointerstitial injury of patients with Balkan endemic nephropathy. Not established the existence of cancer of the upper urinary tract. Macroscopic examination at autopsy of experimental animals, did not determine significant changes in the kidneys. There is first an enormous increase in diuresis in the first and second week of follow-up, in the second and third experimental groups retrospectively, that after 7 weeks of research, diuresis in all test groups was lower than the control group. There is an increase of urea at the end of the research, which is twice higher in the third experimental group compared to the control. There is a marked decrease in uric acid at the end of the research in the experimental group 3. There is a marked decrease in granulocytes in the leukocyte formula in all test groups, and the highest in the third test group. As the decline in the relative values of granulocytes, so there has been a rise in the relative values of lymphocite in the first and second test group. In the third test group, granulocyte drop was accompanied by a extremely large increase in the relative number of basophils. There is a significant drop in specific gravity of urine at the end of the research in the second and third experimental group. Proteinuria is a common finding to all experimental groups, while it was absent or only in traces in the control group. At the end of the experiment was determined to increase significantly the number of phosphate crystals in the experimental groups. The cylinders have appeared only in the urine in the third test group. The greatest number of changes in the urine is determined in the third experimental group.
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Ardin, Maude. "Investigating cancer aetiology through the analysis of somatic mutation signatures." Thesis, Lyon, 2016. http://www.theses.fr/2016LYSE1236/document.
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Les cellules cancéreuses sont caractérisées par des altérations de l'ADN causées par des facteurs exogènes, comme l'exposition à des agents environnementaux tels que le tabac ou les UV, ou par des mécanismes endogènes tels que les erreurs de polymérase lors de la réplication de l'ADN. L'analyse des causes et des conséquences de ces altérations permet de mieux comprendre les facteurs et mécanismes à l'origine du développement d'un cancer. Les technologies de séquençages à haut débit offrent l'opportunité d'étudier la nature précise de ces altérations à l'échelle du génome et permettent de révéler des signatures mutationnelles distinctes et spécifiques de cancérigènes, fournissant ainsi des hypothèses sur l'étiologie des cancers.L'objectif de ma thèse a consisté à développer des méthodes et des outils bioinformatiques accessibles et conviviaux permettant de faciliter l'analyse et l'interprétation des signatures mutationnelles à partir de données de séquençage à haut débit. L'application de ces outils et méthodes à des séries originales de tumeurs humaines et de systèmes expérimentaux de mutagénèse et carcinogénèse a permis de mieux caractériser la signature mutationnelle de l'acide aristolochique (AA) ainsi que d'autres cancérigènes d'intérêtCellular genomes accumulate alterations following exposures to exogenous factors, like environmental agents such as tobacco smoking or UV, or to endogenous mechanisms such as DNA replication errors. Analysing the causes and consequences of these changes allows a better understanding of the mechanisms underlying cancer development and progression. Next-generation sequencing (NGS) technologies provide the opportunity tostudy the nature of the resulting alterations on a genome-wide scale and started to reveal distinct mutational signatures specific to past carcinogenic exposures providing clues on cancer aetiology.The aim of my thesis was to develop user-friendly bioinformatic tools and methods for facilitating the analysis and interpretation of carcinogen-specific mutational signatures from NGS data. Applying these tools and methods to human tumours and experimental models of mutagenesis led to a better characterisation the mutational signature of aristolochic acid (AA), as well as other carcinogens of interest
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Chan, Wan. "Development and application of liquid chromatography and electrospray-ionization mass spectrometry methods for herbal medicine analysis and for the studies of metabolism, DNA adducts and metabonomics of aristolochic acids." HKBU Institutional Repository, 2007. http://repository.hkbu.edu.hk/etd_ra/891.
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Dejan, Miljković. "Histomorfološke, imunohistohemijske i biohemijske karakteristike oštećenja bubrega kod miševa u modelu toksične nefropatije izazvane aristolohičnom kiselinom I." Phd thesis, Univerzitet u Novom Sadu, Medicinski fakultet u Novom Sadu, 2019. https://www.cris.uns.ac.rs/record.jsf?recordId=108220&source=NDLTD&language=en.
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Uvod: Aristolohična kiselina I je nefrotoksična i kancerogena supstanca koja je odgovorna za nefropatiju koja nastaje usled korišćenja herbalnih preparata i čajeva za mršavljenje. S obzirom da se ova supstanca može naći u korovskim biljkama, smatra se jednim od glavnih ekotoksikoloških uzroka za nastanak balkanske endemske nefropatije čiji definitivan uzrok još uvek nije otkriven. Toksičnost ove supstance je dokazana na brojnim animalnim modelima, međutim mehanizmi koji dovode do oštećenja bubrežnog parenhima još u potpunosti nisu razjašnjeni. Cilj: Doktorska disertacija je koncipirana sa ciljem da se utvrdi uticaj toksičnog jedinjenja aristolohične kiseline I na histopatološke i imunohistohemijske karakteristike tubulointersticijuma i glomerula bubrega kod miševa, kao i na biohemijske parametre krvi i urina koji ukazuju na oštećenje bubrega. Materijal i metode: U ekperimentu je korišćeno 64 miša soja NMRI koji su podeljeni u tri grupe: eksperimentalna grupa (n=32) koja je dobijala aristolohičnu kiselinu I rastvorenu u polietilen glikolu (2,5% PEG 400) u dozi od 10 mg/kg telesne mase, negativna kontrolna grupa koja je dobijala 2,5% PEG 400 (n=16) i kontrolna grupa koja je dobijala fiziološki rastovor (n=16). Sve životinje su tretirane intraperitonealno svakodnevno tokom sedam dana. Tokom eksperimenta 8., 17., 29. i 59. dana sakupljan je dvadesetčetvoročasovni urin 8 životinja iz eksperimentalne grupe, 4 životinje iz negativne kontrolne i 4 životinje iz kontrolne grupe. Životinje su žrtvovane 9., 18., 30. i 60. dana, uzeta im je krv, dok su bubrezi posebno odvojeni radi histopatološke analize. Na bubrežnom tkivu sprovedene su histohemijske, imunohistohemijske i morfometrijske analize, dok su na uzorcima seruma i urina sprovedene biohemijske analize. Dobijeni rezultati su testirani adekvatnim statističkim metodama i prikazani su tabelarno i grafički. Rezultati: Nefrotoksin aristolohična kiselina I nakon 7 dana aplikacije izaziva značajno oštećenje bubrežnog parenhima. Pri aplikaciji 2,5% PEG 400 i fiziološkog rastvora ne dolazi do vidljivog oštećenja bubrežnog parenhima. Histopatološku sliku u ranoj fazi eksperimenta (9. i 18. dan) karakteriše akutna tubulska nekroza proksimalnih tubula. U kasnijoj fazi (30. i 60. dana) uočava se histopatološka slika hroničnog intersticijalnog nefritisa sa obilnim mononuklearnim ćelijskim infiltratima limfocitnog porekla kao i postojanje blage intersticijalne fibroze. Kod eksperimentalnih životinja je morfometrijskim metodama utvrđen veći stepen bubrežnog oštećenja tubulointersticijuma i smanjen broj podocita u glomerulu u odnosu na kontrolne grupe. Biohemijske analize kod većine eksperimentalnih životinja su pokazale veće koncentracije serumske uree nego kod kontrolnih grupa. Takođe je dokazana albuminurija u kasnijoj fazi eksperimenta koja je veća kod životinja izloženih aristolohičnoj kiselini I nego kod životinja iz kontrolnih grupa. Zaključak: Korišćenjem morfometrijskih metoda u okviru histopatoloških i imunohistohemijskih ispitivanja, uz adekvatne biohemijske analize, može se zaključiti da je aristolohična kiselina I izuzetno nefrotoksično jedinjenje koje izaziva izrazite promene tubulointersticijuma i glomerula. Podaci ovog istraživanja predstavljaju polaznu osnovu za dalja istraživanja dijagnostike u ranoj fazi nefropatija izazvanih aristolohičnim kiselinama. Introduction: Aristolochic acid I is a nephrotoxic and carcinogenic substance responsible for nephropathy caused by the use of herbal preparations and teas for slimminng regimen. Since this substance can be found in plants, it is considered one of the major ecotoxicological causes for the emergence of balkan endemic nephropathy whose definitive cause has not yet been revealed. The toxicity of this substance has been proven on numerous animal models, but pathophysiological mechanisms of kidney injury still remain unclear. Aim: The doctoral dissertation was designed to determine the influence of aristolochic acid on the histopathological and immunohistochemical characteristics of tubulointerstitium and glomerulus in mice, as well as the biochemical parameters of blood and urine that indicate kidney injury. Material and methods: For this study, 64 mouse of NMRI strain is used. They are divided into three groups: an experimental group (n=32) that received aristolochic acid I dissolved in polyethylene glycol (2.5% PEG 400) at a dose of 10 mg/kg of body weight, a negative control group that received 2.5% PEG 400 (n=16) and a control group that received only saline (n=16). All animals were treated intraperitoneally daily for seven days. During the experiment on the 8th, 17th, 29th and 59th day, twenty-four-hour urine was collected from 8 animals from the experimental group, 4 animals from the negative control and 4 animals from the control group. Animals were sacrificed on the 9th, 18th, 30th and 60th days, their blood was taken, while the kidneys were taken for histopathological analysis. Histochemical, immunohistochemical and morphometric analyzes were performed on renal tissue, while biochemical analyzes were performed on serum and urine samples. Obtained results were tested with adequate statistical methods and presented in a tables and graphs. Results: After 7 days of application nefrotoxin aristolochic acid I causes significant kidney injury. After application of 2.5% PEG 400 and saline, there was no visible damage to kidney parenchyma. Histopathological changes at the early stage of the experiment (9th and 18th day) were characterized by acute tubular necrosis of proximal tubules. At a later stage (30th and 60th day), chronic interstitial nephritis was observed in kidneys, with abundant mononuclear cell infiltrates in interstitium and presence of mild interstitial fibrosis. In experimental animals, a higher tubulointerstitial score of kidney injury and a decrease in the number of the podocytes in glomerulus were determined by morphometric methods, compared to the control groups. Biochemical analyzes in most experimental animals showed higher blood urea nitrogen concentrations than in control groups. High concentration of albumin in urine can be found in later stages of the experiment, and those concentrations were higher in animals exposed to aristolochic acid I than in animals from control groups. Conclusion: Using morphometric, histopathological and immunohistochemical methods, with adequate biochemical analysis, aristolochic acid I is proven to be an extremely nephrotoxic compound that causes drastic changes in tubulointerstitium and glomeruli of kidney parenhyma. Data from this study can be used for further research into early diagnosis of aristolochic acid nephropathy.
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Debelle, Frédéric. "Modèle expérimental de fibrose rénale interstitielle induite par les acides aristolochiques (plantes chinoises)." Doctoral thesis, Universite Libre de Bruxelles, 2005. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/211066.
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La néphropathie aux plantes chinoises (CHN) est une maladie rénale grave qui a été décrite pour la première fois en 1993 chez des patientes ayant suivi un régime amaigrissant à base d’extraits de plantes chinoises (Aristolochia fangchi) contenant des acides aristolochiques (AA). Cette néphropathie se caractérise par une atrophie tubulaire et une fibrose interstitielle aboutissant à l’urémie terminale et se complique fréquemment de cancers des voies urinaires. Au moment d’initier ce travail, il subsistait toujours un large débat quant au rôle étiologique réel des acides aristolochiques dans la genèse de cette maladie. En effet, les gélules à visée amaigrissante contenaient d’autres substances potentiellement néphrotoxiques. Mais surtout, il n’existait aucune preuve expérimentale que les AA pouvaient induire une fibrose rénale interstitielle.Dans la première partie de ce travail, nous démontrons que l’injection par voie sous-cutanée d’AA à la dose de 10 mg/Kg/jour à des rats Wistar mâles en déplétion sodée entraîne l’apparition au 35ème jour d’une atrophie tubulaire, d’une fibrose interstitielle et d’une insuffisance rénale, reproduisant ainsi les anomalies caractéristiques de la CHN. Nous avons ensuite montré que la dexfenfluramine, substance anorexigène à action de type sérotoninergique prise concomitamment par les patientes atteintes de CHN, ne potentialise pas la toxicité rénale des AA. Enfin, la stimulation du système rénine angiotensine (SRA) par la déplétion sodée ou l’inhibition de celui-ci par un traitement pharmacologique ne modifie pas la fibrose interstitielle ni l’insuffisance rénale induite par les AA.
En conclusion, nous avons réussi à développer un modèle in vivo de fibrose rénale interstitielle induite par les AA. Dès lors nous avons apporté la preuve expérimentale de l’implication des AA dans le développement de la CHN. Ce modèle a permis de démontrer que les autres éléments potentiellement néphrotoxiques contenues dans la cure d’amaigrissement (dexfenfluramine, diurétique, laxatif) n’influençaient pas l’évolution de la fibrose interstitielle, ce qui confirme que la prise isolée d’AA suffit à expliquer le développement de la CHN. Cette confirmation à d’importantes implications en santé publique dans la mesure où des plantes contenant des acides aristolochiques font toujours partie des phytothérapies traditionnelles. De plus, il est apparu que, dans ce modèle, les mécanismes de la fibrose rénale interstitielle pouvaient être largement indépendants du SRA. Enfin, de par sa durée limitée et sa grande reproductibilité, ce modèle constitue un outil expérimental d’avenir pour l’étude des mécanismes physiopathologiques de la fibrose rénale interstitielle en général.
Doctorat en sciences médicales
info:eu-repo/semantics/nonPublished
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LI, TSUNG-HSIEN, and 李宗憲. "The aristolochic acids content of Aristolochia zollingeriana leaves and its effect on oviposition of Pachliopta aristolochiae interpositas." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/xsb76b.
Full textAbstract:
碩士國立臺東大學
生命科學研究所
96
Pachliopta aristolochiae interpositas Fruhstorfer prefers to lay eggs on young leave of the food plant, Aristolochia zollingeriana Miq. We suggest that the young leave content more ovipositing attractant, aristolochic acids than older leave. This study compared aristolochic acid content in different stages of A. zollingeriana leaves, and discussed their influence on the oviposition response of P. aristolochiae interpositas. We measured aristolochic acid content in three different stages of A. zollingeriana leaves by HPLC. Total aristolochic acid and aristolochic acid I showed significant differences among the different stages of A. zollingeriana leaves. Furthermore, to examine the oviposition response of female butterfly, we put different treated leaves into a test box, and then put a single female to lay in a test box. After 10 minutes we counted egg number on the leaves of each test. P. aristolochiae interpositas showed no preference to lay eggs on the old leave and the non-host plant leave, however after brushing aristolochic acid on them the egg number obviously increased. In conclusion, aristolochic acid could induce the oviposition response of P. aristolochiae interpositas, and the different aristolochic acid content in the different stages of A. zollingeriana leaves also affected the egg number of P. aristolochiae interpositas laid on them. The preferred stage of A. zollingeriana leaves which female butterflies choose to oviposite on it would affect their larval growth and survival.
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Sun, Chi-Yuan, and 孫啟原. "The investigation on the quantity of Aristolochic Acids-I and-II in the plants of Aristolochia genus and the constituents from fruiting body of Taiwanofungus camphoratus by 1H-NMR and HPLC methods." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/45432588585049703694.
Full textAbstract:
碩士國立成功大學
化學系碩博士班
97
The determination and quantification of aristolochic acids I and II, which will cause acute renal failure, in the plants of the genus Aristolochia is important. In the present study, a quantitative determination of aristolochic acids I and II in plants using 1H NMR spectroscopy was reported. The quantification was performed by relative integration of two proton singlets resonating at δ 8.45 of anthracene. The detection limit was determined to be 0.0107 mg/mL. The 1H NMR spectroscopy is a fast analytical method and relatively easy to perform, but lack the sensitivity that can be achieved with HPLC. Thus, the accuracy of this method was ensured by HPLC. The second part is a method, which was developed for quantitative determination of triterpene acids and benzenoids from the fruiting body of Taiwanofungus camphorates, based on HPLC with UV detector. The HPLC fingerprint of the nine triterpene acids and four benzenoids were developed for the first time and applied to distinguish the wild and cultured species of T. camphoratus. Furthermore, the quantifications of triterpene acids and benzenoids were accomplishment for the wild and cultured species.
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Lai, Yu-Shan, and 賴玉珊. "The effects of aristolochic acids as oviposition and feeding attractants for Pachliopta aristolochiae interpositas." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/5h73wy.
Full textAbstract:
碩士國立臺東大學
生命科學系碩士班
97
Pachliopta aristolochiae interpositas Fruhstorfer distinctly lays more eggs on young leave than on older leave of the foodplant Aristolochia zollingeriana Miq.. The different leaf stages of A. zollingeriana have different kinds and contents of aristolochic acids. The contents of aristolochic acid-
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Lee, Chia-Wen, and 李嘉雯. "Part I: Determination of oregonin in Alnus plants and biological samples by capillary electrophoresisPart II: Determination of aristolochic acid I and aristolochic acid II in Aristolochia debilis and biological samples by UPLC-MS/MS." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/34938728710308769998.
Full textAbstract:
碩士國立臺灣大學
藥學研究所
96
Abstract Part I: Determination of oregonin in Alnus plants and biological samples by capillary electrophoresis Oregonin, existing primarily in the Alnus plants, displayed anti-inflammatory and antioxidative activities. The capillary zone electrophoresis (CZE) method was developed in this study to quantitatively determine oregonin content in the Alnus plants and biological fluid for the first time. Various parameters, including buffer concentration, pH and applied voltage, were evaluated for their optimum analytical conditions. The optimized buffer was composed of 30 mM sodium tetraborate at pH 8.0. The separation voltage was set at 30 kV and the UV detection wavelength was set at 220 nm. Oregonin could be determined within 6 minutes under such optimized conditions. Relative standard deviation (RSD) of the run-to-run repeatability and intermediate precision of the retention time of oregonin was within 1.36 %. Run-to-run repeatability and intermediate precision of the peak area ratios of oregonin to internal standard, theophylline, were both within 1.55 % RSD. The presented method was applied to analyze oregonin in leaves of of Alnus formosana, seeds of various Alnus plants as well as biological samples. The stability of oregonin in biological system was indicated in this study. It demonstrates the potential of this developed method in natural product research. Part II: Determination of aristolochic acid I and aristolochic acid II in Aristolochia debilis and biological samples by UPLC-MS/MS Aristolochic acids (AAs), existing primarily in the Aristolochia plants, shown potent nephrotoxicity and carcinogenicity. AAs are a mixture of structure related compounds, where aristolochic acid I (AA I), and aristolochic acid II (AA II) are reported to be correlated with Chinese herbs nephropathy (CHN). An UPLC-MS/MS method was developed in this study to quantitatively determination of aristolochic acid I, and aristolochic acid II in Aristolochia debilis and biological fluid for the first time. By using pH3.0, 10mM ammonium formate buffer and acetonitrile as mobile phase, Aristolochic acids could be determined within 10 minutes. The mass parameters including cone voltage, capillary voltage, collision energy, ion source temperature, desolvation temperature, desolvation gas flow and the probe distance were subsequently optimized. Selected reaction monitoring (SRM) method was used for quantitative analysis. Relative standard deviation (R.S.D.) of the run-to-run repeatability and intermediate precision of the retention time of AAI and AAII was within 1.25%. Run-to-run repeatability and intermediate precision of the peak area of AAI and AAII were both within 5.74% R.S.D. Accuracy of the method was between 98.11% and 114.51%. Limit of detections was 0.14 ng/ml for AAI and 0.26 ng/ml for AAII. The result shows that the present UPLC-MS/MS method is efficient, accurate and sensitive for the determination of AAI and AAII.
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Huang, Szu-Ping, and 黃思萍. "An exposure biomarker of aristolochic acid-containing herbs: aristolochic acid and N-acetyl-L-cysteine adduct." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/6ewhgk.
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Tseng, Chiachi, and 曾家琪. "Acute Nephropathy of Aristolochic Acid in Rabbits." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/01003885275294091901.
Full textAbstract:
碩士臺北醫學大學
藥學系
94
Chinese herbs induced nephrotoxicity has spread worldwide recently. Inadvertly misuse of the Chinese herbs, which contain aristolochic acids （AA）, is the main cause of the nephropathy. Aristolochic acid nephropathy （AAN）is characterized by its extensive tubular atrophy with rare cell infiltration and interstitial fibrosis while glomeruli are relatively spared. In the present study, renal toxic effects of AA of various doses or different routes were evaluated through renal histology and function in rabbits. AA was intravenously injected (IV) into rabbits with various doses：0.25, 0.5, 1.0 and 2.0 mg/kg to rabbits. AA was also given orally （PO）with 1.0 mg/kg to rabbits. After dosing, rabbits were under monitoring of plasma creatinine, serum N-acetyl-beta-D-glucosaminidase（NAG）activity and sacrificed on different days for the histological examination. After the previous study, AA 0.25 mg/kg was administrated intravenously again to understand the possibility of tubular regeneration after AA intoxication. Plasma creatinine increased immediately after AA treatment in all conditions. The results of plasma creatinine and serum NAG monitoring indicated that renal function was changed slightly after IV 0.25 mg/kg and PO 1.0 mg/kg AA while higher doses of aristolochic acid had greater impacts on renal function. Serum NAG was less sensitive than plasma creatinine and not suitable for monitoring renal function. Histologically, tubular atrophy was significant in all conditions and AA developed widespread tubular necrosis in higher doses. There was no remarkable evidence of tubular regeneration observed in this study. The glomeruli were relatively spared. There were good relationships between biochemistry and tubulointerstitial histological scores（TIHS）when renal was impaired severely. After 2.0 mg/kg AA intoxication, biochemistry correlated tightly with tubulointerstitial scores was observed （P < 0.05）.In other doses, the elevations of plasma creatinine levels were followed by higher THIS. In conclusions, the renal toxic effects of AA were significantly dose-dependent（P < 0.05）.
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Wei, Yu-Yun, and 魏玉雲. "Genotoxicity of aristolochic acid in human cell lines." Thesis, 2003. http://ndltd.ncl.edu.tw/handle/66617783032372637791.
Full textAbstract:
碩士國立陽明大學
環境衛生研究所
91
Aristolochic acid (AA), derived from Aristolochia spp., was related to the development of a novel nephropathy and urothelial carcinomas in patients with aristolochic acid nephropathy (AAN). Since treatment of mammalian cells with AA—a mixture of AAI and AAII─has been shown to result in induction of DNA mutagenesis and carcinoma in animals, this study is designed to investigate whether genotoxicity can be induced in AA-treated human skin fibroblast (HSF) and 1BR3 cells. After various AA exposure, HSF and 1BR3 cells were evaluated for the frequencies of nuclear foci by labeling with anti g-H2AX monoclonal antibody, which can detect repair proteins for DNA double-strand breaks, as well as micronuclei and p53 protein. Exposure of AA led to significant decrease in viabilities of both HSF and 1BR3 cells in dose- and time-dependent manners. Slight time-dependent, but not dose-dependent, increases in proportions of g-H2AX nuclear foci in both HSF and 1BR3 cells were shown after AA exposure. With the frequencies of g-H2AX nuclear foci are 0.77-1.77% in HSF cells and 0.58-2.31% in 1BR3 cells. Dose-dependent increase in the micronuclei frequencies (MF) was shown in HSF cells. MF in the binucleated HSF cells induced by cytochalasin-B was higher than those observed in mononucleated ones. HSF cells post AA was shown with increase in total p53 proteins. Furthermore, the levels of p53 were found to be dose- and time-dependent in HSF cells after AA exposure. There was a high association between MF and p53 proteins (R>0.84) in the dose-dependent manner. These indicated that AA is with genotoxic and/or mutagenic effects in human cells, and both MF and p53 protein were more sensitive for the evaluation. The association between DNA adducts and other DNA damages induced by AA in human cells remains to be further established.
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Fan, Mei-Yu, and 范美玉. "Pharmacokinetics and nephrotoxicity of aristolochic acid in rabbits." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/93046977656572245429.
Full textAbstract:
碩士臺北醫學大學
藥學系
94
Aristolochic acid (AA) is the main component of Aristolochia plants. It is a mixture of aristolochic acid Ⅰ(AAⅠ) and aristolochic acid Ⅱ(AAⅡ). It has been proved by several researches that AA has nephrotoxicity and tumor toxicity. However, the pharmacokinetics of AA is still limited. A validated high performance liquid chromatography (HPLC) method was used to determine the plasma concentration of AAⅠ and AAⅡ. The HPLC method used was met the guidance of bioanalytical method validation. The pharmacokinetics of AAⅠ and AAⅡ was studied by respectively intravenous administration of four different doses(0.25, 0.5, 1.0, 2.0 mg/kg ) of aristolochic acid sodium salt (AANa) in four groups of rabbits (n=6 for each group). Because of detection limit, the results of AAⅠ and AAⅡ from 0.25 mg/kg AANa were fit with one compartment model. Others were fit with two compartment model. There was no significant difference (P>0.05) in pharmacokinetic (PK) parameters of AAⅠ and AAⅡ, except 0.25 mg/kg AANa-treated group. The system clearance of AAⅠ were 349.27112.92, 236.9948.92, 228.4731.04, 226.9919.42 mL/h/kg. Clearance for AAⅡ were 224.2034.01, 199.5841.93, 156.7630.30, 185.4229.81 mL/h/kg. The area under the curves(AUC) of AAⅠ and AAⅡ were calculated from zero to infinite. The AUCs of AAⅠ and AAⅡ were proportional to the dose administrated (AAⅠ: Y= 4754.23X-188.20, r2=0.980, P<0.001; AAⅡ: Y= 5728.56X-21.57, r2=0.911, P<0.001). It indicated that AAⅠ and AAⅡ might behave dose-independent pharmacokinetics between 0.25~2.0mg/kg of AANa IV injection. In another study, six rabbits were intravenous administered with different doses (0.5, 1.0, 2.0 mg/kg) of AANa. All animal of this study were treated with AANa with 7 days interval. The PK parameters of AAⅠ and AAⅡ were changed between 0.5 ~ 2.0 mg/kg of AANa IV injection. The clearance of AAⅠ were 236.9948.92, 198.3755.09, 102.1022.39 mL/h/kg. The clearance of AAⅡ were 199.5841.93, 97.6323.22, 42.317.64 mL/h/kg. The AUCs of AAⅠ and AAⅡ were not proportional to the increasing doses. The relationship of non-linear pharmacokinetic properties between dose and AUC were obtained (AAⅠ: Y=13618.09X2 - 875.43X + 469.52, r2=0.908, P<0.001; AAⅡ: Y=26583.73X2 - 3779.03X + 489.38, r2 = 0.948, P< 0.001 ). The non-linear pharmacokinetics might due to renal lesions caused by AAⅠ and AAⅡ. After oral administration of AANa 1.0 mg/kg, the gastric acid would destroy AA Ⅰand AAⅡ. The amount of AAⅠ and AAⅡ detected in plasma were very low. And the absorption of AAⅠand AAⅡ was not regularity and in high variation. After treated with AANa, rabbits were sacrificed on day-1 and day-8 to obtain the kidney sections. The histological examination of intravenous administration of AANa was described as the following. On day-1, there was no significant alteration in renal morphology on 0.25 mg/kg AANa-treated group. Mild interstitial infiltration, epithelial cell degeneration and atrophy were observed on 0.5, 1.0, and 2.0 mg/kg AANa-treated group. On day-8, mild degeneration of epithelium and atrophy were observed on 0.25 mg/kg AANa-treated group. Moderate to severe renal lesions including, interstitial infiltration, degeneration and atrophy of epithelium, interstitial fibrosis and hyaline cylinders were found on 0.5, 1.0 and 2.0 mg/kg group. Kidney sections were also obtained on day-1 and day-8 after oral treated with 1.0 mg/kg AANa to rabbits. In oral study, there was no significant change in renal morphology on day-1. But mild degeneration and atrophy of epithelium and interstitial fibrosis were observed on day-8. According to the histological examination, the extent of renal lesions increased with doses increasing was elucidated. The renal lesions caused by AA were continued to deteriorate. However, lesions in glomerulus were not observed.
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Tsai, Dong-Ming, and 蔡東銘. "Metabolomics Study of Aristolochic Acid Nephrotoxicity in Rodents." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/91931900984537061181.
Full textAbstract:
碩士國立臺灣大學
生醫電子與資訊學研究所
97
Aristolochic acid (AA) is a potent nephrotoxic agent that can be found in several herbs of the genus Aristolochia. It was once commonly used in traditional Chinese medicine remedy. In this study, we applied our metabolomics platform of the 1H NMR spectroscopy on urine with principal component analysis (PCA) to detect and further dissect the nephrotoxicity in rodent of four AA containing materials, AA standard, Madouling, an AA containing herb Aristolochia contorta and Bu-Fei-A-Jiau-Tang (BFAJT), a herb compound containing Aristolochia contorta. The aim was to use the easy available urine samples to establish a model for early diagnosis of nephrotoxicity. The experiment was divided into four parts. The first was rat AA experiment. The second was mouse AA experiment. The third was mouse Madouling experiment. The fourth was mouse BFAJT experiment. Urine samples were collected daily and freeze-dried for storage. All animals were euthanized after experiment and their kidneys and livers procured for pathological studies. Results: In the rat AA experiment, pathology showed grade (gr.) 1-3 acute renal tubulointerstitial necrotic change (ATIN) after 5 doses in the high dose group. In the mouse AA experiment, 10 doses were given. Both high and low dose group showed gr. 3-4 ATIN. In the mouse Madouling experiment, 21 doses were given. It was gr. 1-2 for low and moderate groups, and gr. 3-4 ATIN for the high dose group. In the BFAJT experiment, 20 doses were given. It was gr. 2 ATIN in the low dose group and gr. 3-4 in the high dose group. PCA scoring plots for the urine NMR spectral chemical shifts variables showed early cluster at 2 day and later for each group in the rat AA experiment. In the mouse AA experiment, the high dose group was clustered from the other 2 groups at day 8. It was at day 10 that the 2 dose groups were clustered from the control group. In the mouse Madouling experiment, the high dose group clustered from all other 3 groups. It was at day 10 that the high dose and the moderate dose groups clustered together from the other 2 groups. In the BFAJT experiment, PCA scoring plots failed to classify across control and dose groups even at day 16. The endogenous metabolites assigned from NMR spectroscopy and their integral concentration among dose and control groups were tested by t-test. In the rat AA experiment glycine, succinate, 2-oxoglutarate and trimethylamine-N-oxide (TMAO) were decreased in the dose groups in earlier days of the experiment. At later days, sugar, allantoin, creatine, dimethylglycine, 2-oxoglutarate and TMAO were increased in the dose groups (p < 0.05). In the mouse AA experiment no significant metabolite difference was noted, only alanine, lactate and formate had more than two fold change at day 10. In conclusion, AA standard, Madouling and BFAJT were all nephrotoxic. PCA scoring plots for the urine NMR spectroscopy collected from living animals showed the ability to classify the dose and control groups days before confirming the renal pathology. But toxicity classifying failed in the mouse BFAJT experiment by PCA scoring plot. Further metabolomics study is expected to resolve this issue.
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Ming-Feng, Huang. "Capillary electrophoresis for the analyses of amines, aristolochic acids and DNA." 2006. http://www.cetd.com.tw/ec/thesisdetail.aspx?etdun=U0001-2607200605295600.
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Huang, Ming-Feng, and 黃銘峰. "Capillary electrophoresis for the analyses of amines, aristolochic acids and DNA." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/03563470713272948163.
Full textAbstract:
博士國立臺灣大學
化學研究所
94
Abstract This thesis focuses on developing efficient and sensitive capillary electrophoresis-laser induced fluorescence (CE-LIF) techniques for amines, aristolochic acid, and DNA. We developed a novel method for the analysis of amines under acidic conditions by CE in conjunction with indirect laser-induced fluorescence (CE-ILIF). The analysis of six amines by CE-ILIF using a solution, pH 3.5, containing 5.0% methanol, 0.1 mM sulfuric acid, 0.1 mM cresyl violet, and 0.3 mM lithium was complete in 5 min, with the limits of detection (LOD) on the level of µM.To further improve the sensitivity, on-line concentration based on pH junction has been demonstrated. When injecting the sample prepared in a solution of 0.2 mM sulfuric acid, pH 3.3, at 15 kV for 60 s to the above-mentioned solution, the LODs for the amines down to sub µM and the sensitivity improvements up to 19-fold when compared to that injecting at 15 kV for 5 s. Aristolochic acid (AA), a naturally occurring nephrotoxin and carcinogen, has been associated with the development of urothelial cancer in humans. Two predominant forms of AA are 8-methoxy-6-nitrophenanthro-(3,4-d)-1,3-dioxolo-5- carboxylic acid (AA-I) and 6-nitro-phenanthro-(3,4-d)-1,3-dioxolo-5-carboxylic acid (AA-II). Owing to lack of intrinsic fluorescence characteristics of oxidized AAs (OAAs), the RAAs reduced from OAAs in 10.0 mM HCl containing iron powder is required prior to CE analysis. The RAAs exhibit fluorescence at 477 nm when excited at 390 nm. By using 50.0 mM sodium tetraborate (pH 9.0) containing 10.0 mM SDS,the determination of aristolochic acid I and aristolochic acid II by CE-LIF has been achieved within 12 min. The CE-LIF provides the LODs of 8.2 and 5.4 nM for AA-I and AA-II, respectively. The successful analysis of 61 medicinal samples and dietary supplements shows that the present CE-LIF is practical for the determination of AA-I and AA-II in real samples. Reproducible, rapid, and high-resolution DNA separations have been achieved using low-viscosity PEO solutions containing GNPs ranging in diameter from 3.5 to 56 nm. The separation of DNA ranging in size from 8 to 2176 base pairs (bp) was accomplished in 5 min using 0.2% PEO(8 MDa) containing 56-nm GNPs. We have also demonstrated the separations of the DNA fragments ranging from 5 to 40 kbp using 0.05% PEO(2 MDa) containing 13-nm GNPs or 0.05% PEO(4 MDa) containing 32-nm GNPs. To separate long double strands of DNA by CE, with high efficiency, high speed, simplicity, and reproducibility, we describe a CE technique, which we call nanoparticle-filled CE (NFCE), using polymer-modified gold nanoparticles (GNPPs) in this topic. The gold nanoparticles were modified with poly(ethylene oxide) via noncovalent bonding to form GNPPs. The separations of high molecular weight (HMW) DNA with sizes ranging from 8.27 to 48.5 kbp and λ DNA (0.12–23.1 kbp) were accomplished within 6 and 5 min, respectively. The separation speed and resolution are greater than those by pulsed CE and slab gel electrophoresis. This is the first example for separating such high DNA molecules by CE-LIF using GNPPs. The results present in this thesis clearly demonstrate that CE-LIF based techniques are practical for analysis of amines, aristolochic acid, and DNA, with the advantages of rapidity, sensitivity, and reproducibility. The examples of separating DNA using PEO containing GNPs and GNPPs open the avenue of using nanoparticles for improved resolution and reproducibility for analysis of DNA. It is our strongly belief that the technique should be further applied to analysis of small solutes such as amines and acids and macromolecules like proteins.
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Liu, Chiung-Yueh, and 劉瓊月. "Mechanisms Involved in the Antiplatelet Activity of Aristolochic Acid." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/85103362180460893842.
Full textAbstract:
碩士臺北醫學大學
醫學研究所
94
Aristolochic acid (AsA) is an alkaloid from the plant Aristolochiaceae. The naturally occurring herbal toxin that can cause cancer and end-stage kidney failure. Recently, it had been reported that AsA could reduce edema induced by snake venom and possess anti-inflammation and anti-platelet activity of AsA. However, the mechanisms involved in anti-platelet activity of AsA is still unclear, and we are interested in investigating the effects on cellular signal transduction during the process of platelet activation. In this study, AsA concentration-dependently (75-150 micromolar) inhibited collagen (1 microgram/ml) induced human platelet aggregation and ATP release reaction. AsA (115 and 150 micromolar) inhibited intracellular Ca2+ mobilization, phosphoinositide breakdown, and thromboxane A2 formation stimulated by collagen (1 microgram/ml) in human platelets. In addition, AsA (115 and 150 micromolar) increased levels of nitrate and induced phosphorylation of vasodilator-stimulated phosphoprotein (VASP). Phosphorylation of 47 kDa proteins is a marker of protein kinase C activation, and can be triggered by collagen (1 microgram/ml) and PDBu (150 nano molar). In our experiments, we found AsA (115 and 150 micromolar) could inhibit phosphorylation of 47 kDa proteins by collagen (1 microgram/ml). Besides, AsA (115 and 150 micromolar) reduced p38 MAPK phosphorylation induced by collagen (10 microgram/ml), and had no effects on scavenging collagen (1 micro gram/ml)-induced hydroxyl radicals in platelets. In conclusion, our study suggested that the pathways of AsA (115 and 150 micromolar) about anti-platelet activity maybe involved the following: (1) AsA could regulate the activity of PLC and then inhibit phosphoinositide breakdown, intracellular Ca2+ mobilization and 47 kDa protein phosphorylation. (2) AsA significantly reduced thromboxane A2 formation maybe through inhibition of p38 MAPK phosphorylation and Ca2+ mobilization, which are responsibe for PLA2 activation. (3) AsA inhibted the activity of platelet by increasing the amount of NO and VASP phosphorylation .
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Yu, Chia-chen, and 余佳真. "Effects of Antioxidants (Vitamin C and E) on Toxicity of Aristolochic Acid and Comparative Toxicity of Aristolochic Acid I and II in Rats." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/76317519694067230997.
Full textAbstract:
碩士國立臺灣海洋大學
食品科學系
93
Since the workshop on herbal medicine poisonings in Asia-Pacific zone and recent references reported that scientific liver-protecting herb and some slimming regimen containing aristolochic acid (AA), the safety of these Chinese herbs and slimming regiment is becoming more concern. These substances could cause Chinese herb nephropathy (CHN), a unique type of rapidly progressive interstitial nephropathy. In addition, these Chinese herbs are usually rich in vitamin C and vitmin E. This study, therefore, was focused on the changes in toxicity of AA with vitamin C and E. Forty-eight experimental rats were divided into six groups, including control (1 ml saline), AA (10 mg AA/1 ml), vitamin C (10 mg vitamin C/1 ml), vitamin E (1.5 mg vitamin E/1 ml), vitamin C with AA (10 mg AA+10 mg vitamin C/1 ml) and vitamin E with AA (10 mg AA +1.5 mg vitamin E/1 ml). Eight Wistar rats of each group were fed by oral treatment. After two months, AA group had significant (P＞0.05) relative ratios of liver and kidney weight to body weight than others. On the other hand, the levels of cholesterol and triglyceride in plasma were not significant different between each group(P＞0.05). The aspartate transaminase (AST), alanine transaminase (ALT) values in plasma of AA group were higher than those of others (P＜0.05), but the alkaline phosphatase (ALP) value was not significant different (P＞0.05). Blood urea nitrogen (BUN) and creatinine in the plasma of AA group were higher than those of others (P＜0.05). The low density lipid (LDL) value in plasma of AA group were also higher than that of others (P＜0.05), but the high density lipid (HDL) value was not significant different (P＞0.05). In conclusion, AA indeedly caused kidney disease and difference of lipid in plasma, but vitamins C and E played the function to reduce the toxicity of AA. Because the toxicity of single component of AA is still unknown, the AA I and AA II were udentified and preparated by HPLC. After that, Wistar female rats were used for this experimental animal. As above method mentioned, the experimental rats were fed with AA I and AA II one time per three days. After two months, we compared the toxicity of AA I and AA II on rats. The result showed that AA I and AA II group were higher than control group in the relative ratio of liver and kidney weight to body weight in rats. AA I and AA II were lower than control group in aspect of red blood cells and hematocrit on the 8th week. There were insignificant difference among the groups in the white blood cells and hemoglobin of plasma. On the other hand, AA I and AA II groups showed insignificant difference, but their results were higher than the control group with respect to the values of kidney functional indicators BUN and creatinine, and the values of liver functional indicator AST and ALT on the 8th week. As to ALP in plasma, they had insignificant difference. The LDL value in plasma of AA I and AA II group were higher than that of control group. Every group showed insignificant difference for the other values in plasma such as cholesterol, tryglyceride and HDL. In summary, the toxicity of AA I and AA II was almost the same. They both caused kidney damage and a little liver damage for Wistar female rats.
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葉世銘. "Removal of Toxic Compound, Aristolochic Acid, by using Carbopol Material." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/67246684021556516846.
Full textAbstract:
碩士明新科技大學
化學工程研究所
96
The main purpose of this study is to remove the toxic compound, Aristolochic Acid, existing in Aristolochia liukiuensis. The scientific and safe biomaterial Carbopol is utilized to remove Aristolochic Acid from natural product to nontoxic or less toxic status. Hence the first step for this research is to develope the methodology for analyzing the amount of Aristolochic Acid existing in Aristolochia liukiuensis. A suitable High Performance Liquid Chromatography (HPLC) method with stationary phase of RP-18 (10 μm) and mobile phase of 66% methanol：34% DI water was developed for Aristolochic Acid identification and quantification. Carbopol, which possesses high molecular weight, is cross-linked water-soluble polyacrylic acids polymer. It was used for the purpose of removing toxic compound, since they both are having polar functional groups. The carboxylic acid functional group of Carbopol was identified by Fourier Transform Infrared Spectrometer (FT-IR). Carbopol, which is available in different viscosity grades, has no effect on the biological activity of the drug while has excellent thickening and suspension effects. Therefore Carbopol polymer is also used in this research for making the control-release formulation. Carbopol and the detoxified natural medicine are blended to prepare gelatinic product. Both stability kinetics and the release pattern were tested for the prepared product. The release rate by disintegration is near 100% after five hours but the dissolution rate is only 10% after three hours. The prepared product is stable under room temperature for five days.
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I-Tzu and 陳怡孜. "Inhibition of LPS-Induced Cellular Inflammation by Phytotoxin Aristolochic Acid." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/09225779948415073428.
Full textAbstract:
碩士中山醫學大學
生物醫學科學學系碩士班
96
Aristolochic acid (AA) is a group of natural compounds widely found in Artistolochia species and is mainly composed of aristolochic acid I (AAI) and aristolochic acid II (AAII). Recently AA is found to be nephrotoxic and carcinogenic to humans and rodents. However, the extract of Artistolochia serving as Chinese herbal medicine has been commonly used to treat arthritis and rheumatism for hundreds of years. In addition, AA compound has the ability to increase phagocytic activity of phagocytes and has potential anti-inflammatory properties. In the present study, we focused on the anti-inflammatory effect of AA on the rat cell line (RAW 264.7) and further studied its reaction mechanism. Lipopolysaccharide (LPS) was used to stimulate RAW 264.7 macrophages to generate a cellular inflammation model. When RAW 264.7 macrophages were treated with LPS, the levels of nitric oxide (NO) production, inducible nitric oxide synthase (iNOS) mRNA and protein expression were all increased. Co-treatment of RAW 264.7 macrophages with LPS and various concentrations of AA led to the down-regulation of LPS-induced NO, iNOS protein and mRNA levels in a concentration-dependent manner. Since AA above 100 μM showed no significant effect on the cellular viability of RAW 264.7, the AA concentration we used can efficiently inhibit LPS-induced cellular inflammation without showing cellular toxicity. The induction of iNOS gene expression is known to be correlated with the regulation of various transcription factors, so both in vitro and in vivo assays were conducted to investigate if AA can affect the DNA binding activity of these transcription factors. By the in vitro Electrophoretic Mobility Shift Assays (EMSA), we found that exposure of LPS-treated cells to AA did not block the NF-κB DNA binding activity mediated by LPS. By using Luciferase reporter assay in the cellular experiment, we observed that the AA treatment did not alter the luciferase activities of reporter plasmids harboring NF-κB, AP-1, ISRE or GAS DNA binding sequences in front of luciferase gene. Besides, the presence of AA showed no influence on the status of LPS-induced STAT-1 phosphorylation and did not significantly inhibit the translocation of NF-κB from cytoplasm to nucleus. However, high concentrations of AA could obviously suppress the phosphorylation of I-κB induced by LPS; I-κB protein is known to be an inhibitor of NF-κB. To further confirm that AA is able to decrease iNOS gene transcription, three luciferase reporter plasmids containing 1588 bp, 1008 bp or 360 bp iNOS promoter sequence from the starting site were constructed. The results demonstrated that AA could apparently decrease the LPS-induced luciferase activities of all the constructed plasmids, suggesting that the inhibitory effect of AA may be associated with an unknown DNA sequence within the 360 bp iNOS promoter.
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Chen, Jiun-Liang, and 陳俊良. "Investigation of the Mechanisms Underlying Aristolochic Acid-Induced Kidney Injury." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/45718352810064632742.
Full textAbstract:
碩士長庚大學
傳統中國醫學研究所
93
Objective: To investigate the mechanism underlying aristolochic acid-induced kidney injury. Both cell model and animal model experiments were carried out to study the in vitro and in vivo effects of aristolochic acid. Methods: In vitro study:Renal tubular epithelial cells ( NRK-52E ) were cultured in 95 % Dulbecco's modified Eagle's medium (DMEM) with 0.1mM non-essential amino acid and 5% calf serum. The cells viability was measured by theMTT(3-[4,5-Dimethylthiazol-2-yl]-2,5-dihenyltetrazolium bromide；Thiazolyl blue) assay. Apoptotic cells were identified by propidium iodide ( PI ) stain, TUNEL(TdT-mediated dUTP nick end labelling) assay and Annexine-V FITC (Fluoreszeinthiocyanat ) stain. RNA expression was analyzed by RT-PCR. In vivo study: The histopathology of FVB mice was examed by hemotoxylin and eosin stain, TUNEL assay and immunohistochemical method. Results: (1)Aristolochic acid in the concentration above 10μg/ml could significantly reduce the viability of NRK-52E rat kidney proximal renal tubule cell. (2) TUNEL stain demonstrated the apoptosis of renal tubule cells induced by aristolochic acid. (3) Annexine-V FITC (Fluoreszeinthiocyanat ) stain further confirmed the apoptosis of renal tubule cells induced by aristolochic acid.(4) Aristolochic acid in the concentration above 10μg/ml could increase the expression of Bax mRNA.(5)Pretreatment of cyclosporin A (7.5μg/ml ) could reduce the apoptosis of renal tubule cells induced by aristolochic acid.(6)Radix salviae miltiorrhizae could decrease the loss of cells by aristolochic acid. was significantly inhibited. Pretreatment with radix salviae miltiorrhizae could prevent the cell apoptosis of kidney proximal renal tubule cells induced by aristolochic acid.(7)The FVB mice model of aristolochic acid nephropathy confirmed the cell apoptosis of kidney proximal renal tubule cells. Conclusions : Aristolochic acid in the concentration above 10μg/ml could significantly reduce the viability of NRK-52E rat kidney proximal renal tubule cell and increase the expression of Bax mRNA via mitochondria pathway to induce the apoptosis of renal tubule cells. Radix salviae miltiorrhizae exerts dose-dependent protective effect on the aristolochic acid-induced renal tubular epithelial cells injury by preventing the apoptosis of renal tubular epithelial cells induced by aristolochic acid. The animal model of aristolochic acid nephropathy confirmed the apoptosis of renal tubular epithelial cells induced by aristolochic acid by TUNEL stain. We have set up the FVB mice model of Chinese herb-induced tubular interstitial nephritis which could help us to screen and study the Traditional Chinese medicines in the prevention or treatment of Chinese herb-induced tubular interstitial nephritis and the early renal fibrosis.
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Martináková, Lenka. "Studium metabolizmu karcinogenní a nefrotoxické přírodní látky aristolochové kyseliny II." Master's thesis, 2019. http://www.nusl.cz/ntk/nusl-396642.
Full textAbstract:
Aristolochic acids (AA) have been considered as toxicants of plants which were found in plants of the family Aristolochiaceae. The most abundant acids in mentioned plants are aristolochic acid I (AAI) and aristolochic acid II (AAII). AA have been considered as causes kidney disease called Aristolochic acid nephropathy (AAN). AAN was initially discovered in patients of one Belgian clinic in Brussels specialized on treatment of patients leading to a decrease in their body weight. The first name of this disease was Chinese herb nephropathy (CHN). Later, it was discovered that one component of herbal preparation was changed by a mistake with the Aristolochiaceae plant. The second type of renal disease caused by AA was discovered in populations of countries along the Danube river, called as Balkan endemic nephropathy (BEN), which was probably caused by the contamination of grains with plants containing AA. These renal diseases (AAN and BEN) are often associated with development of upper urothelial cancer (UUC). AA (AAI + AAII) in organisms are subject to biotransformation leading to its reductive activation or oxidative detoxification. Both cytosolic enzymes [NAD(P)H:quinone oxidoreductase] and microsomal enzymes [cytochromes P450, NADPH:cytochrome P450 reductase] participate in their reduction. The...
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Tseng, Chiao-Shih, and 曾喬詩. "Pharmacokinetic Studies of Inulin and p-Aminohippuric Acid in Rabbits with Aristolochic Acid Nephropathy." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/68546356713676204462.
Full textAbstract:
碩士臺北醫學大學
藥學研究所
95
On morphology of AAN, the proximal tubules were the main target of AA-related nephrotoxicity. It is interesting to note that glomerular structure were not affected. This study was done to speculate renal function changes while AAN presented. This study was performed via a single i.v. administration of 0.5mg/kg AA sodium salt (AANa) for 8 days resulted in the appearance of moderate tubulointerstitial lesions in male New Zealand white rabbits. Taking the grading system to evaluate AA-induced tubulointerstitial lesions, the sum of the histological scores were increase by comparison with normal rabbits. (2.09±1.23 vs. 5.03±1.05; P<0.01). Then 20mg/kg of p-aminohippuric acid (PAH) and inulin was iv administered respectively. The renal plasma flow (RPF) and glomerular filtration rate (GFR) changes were assessed by changes of pharmacokinetics of PAH and inulin. The concentrations of PAH and inulin in plasma was determined by HPLC methods. RPF was calculated as CLPAH/EPAH and EPAH is the PAH excretion ratio. The results showed that effects of 0.5mg/kg AANa treatment on PAH in rabbits were α-t1/2 (P=0.016) and β-t1/2 (P=0.026) significantly increased and CL (P=0.010) and k10 (P<0.001) were significantly decreased. The pharmacokinetic parameters of PAH, AUC (vs. interstitial filtrateion；P=0.006，vs. total histological score；P=0.047) and β-half-life (vs. hyaline cylinders；P=0.017，vs. interstitial fibrosis；P=0.020), were significantly correlated with the levels of tubulointerstitial lesions. Changes in the pharmacokinetic data of PAH might have resulted from the moderate alterations observed in the proximal tubules. Although effects on inulin were CL (P=0.014) and k (P<0.001) significantly decreased, there was no significantly correlation between pharmacokinetic data and tubulointerstitial lesions. Inulin only eliminated via filtration could be a explanation. And RPF with AAN were decreased when compared with controls (22.36±4.01 vs. 10.11±2.72mL/kg/min; P<0.05). For GFR were also decreased (5.95±1.79 vs. 0.89±0.18mL/kg/min; P<0.05). The different proportional fall in GFR (-82.84±2.81%) and RPF (-50.55±13.59%) suggests preglomerular vasoconstriction led GFR changes so much. We also found that tubular anion secretion (33.34 vs. 18.27mL/min) tend to decline. Thus, consideration dosage adjustment in clinical practice not only based on GFR but also the ability of tubular secretion. This study demonstrates that the renal function RPF and GFR were decreased while AA-induced major injure in the proximal tubules.
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Chen, Yen-Chi, and 陳彥祺. "Analysis of Aristolochic Acid in Xin Yi San by HPLC-UV." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/97675156267837145518.
Full textAbstract:
碩士國立臺灣海洋大學
食品科學系
99
Abstract A high performance liquid chromatography (HPLC) for the determination of aristolochic acid in Chinese medicinal herbs has been studied. Xin Yi San was examined in detail and aristolochic acid contents in both ethanol and water extracts were also compared in this study. The samples (Xin Yi San and Asiasarum) were analyzed using the HPLC method on a C-18 column (Cosmosil 5 C 18-AR-II 4.6×250 mm) and were detected at 395nm with acetonitrile and 2% glacial acetic acid aqueous solution (1：1) (V/V), as mobile phase at a flow rate of 1.0 mL/min. This method was a sensitive, economic, scientific and feasible method for measuring aristolochic acid I. All the linear regression equation coefficients were greater than 0.999, detection limits was 0.05 mg. The recoveries of samples were 96.17%~98.65%.
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Ding, Yu-Ju, and 丁玉如. "Screening of the chemopreventive componds against aristolochic acid-induced nephropathy from natural products." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/56600529645286014673.
Full textAbstract:
碩士淡江大學
生命科學研究所碩士班
98
Aristolochic acid (AA) occurs naturally in Aristolochiaceae plants, and toxicological studies have shown the role of AA in human aristolochic acid nephropathy (AAN). Most studies have focused on the adult toxicity of AA, but the embryonic toxicity and toxicological mechanisms are still not clear. Therefore, we used zebrafish embryos to investigate the effects of AA on embryonic development and toxicity. After 24-hpf embryos were treated with AA (10 ppm) for 7 hours, the 48-hpf embryo kidney (glomeruli, pronephric tubules, and pronephric ducts) and heart (deformed heart, swollen pericardial cavity,and hemorrhagic yolk sac) were severely damaged, and the 72-hpf survival rate was decreased to 14.2%. The glomerular filtration rate (GFR) of embryos was further examined after AA treatment. The GFR of embryos after 3-h AA treatment decreased to 71.5 ± 18.8%, while that after 5-h AA treatment decreased to 39.4 ± 15.9%. Taken together, AA caused renal injury and dysfunction, as well as reduced survival rate of zebrafish embryos. Besides, AA-induced apoptosis was observed. A significant decrease in the amount of erythrocytes and leukocytes was observed in AA-treated embryos. Moreover, erythrocytes became stationary in blood flow and even accumulated in renal tubules and intestinal tract. Based on quantitative PCR, mRNA expression of inflammatory genes (TNF-α, cox2, and mpo) was upregulated in AA-treated embryos. 12-h treatment (12-24 hpf) of resveratrol or ursolic acid before AA treatment (24-31 hpf) was found to inhibit AA-induced inflammation and attenuate but not completely reverse AA-induced renal injury. In summary, the cellular and molecular mechanisms of AA toxicity may result in inflammatory renal injury and hematopoietic dysfunction, severe defects of heart and kidney, and ultimately lead to kidney dysfunction and failure.
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Chen, Hung-Hsiang, and 陳泓翔. "Proteomics analysis of the effects of prednisolone on aristolochic acid nephropathy in mice." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/x4g47s.
Full textAbstract:
碩士臺北醫學大學
藥學系(碩博士班)
102
This study was designed to investigate the effects and mechanism of prednisolone treatment in aristolochic acid nephropathy (AAN) mice by proteomics analysis. The male C3H/He mice were separated into normal, AA and prednisolone group and the experimental period was 10 weeks. Both AA group and prednisolone group were treated with AA (0.5 mg/kg/day) for 8 weeks. For the next two weeks, AA group was treated with distilled water while the treatment group was treated with prednisolone (2 mg/kg/day). The normal group was administered distilled water for 10 weeks without AA. The homogenate of the kidney was reacted with DAABD-Cl (4-[2-(Dimethylamino) ethylaminosulfonyl]-7-chloro-2,1,3-benzoxadiazole). The derivatized proteins were separated and quantified by high performance liquid chromatography with fluorescence detection (FD-HPLC). The differential proteins were identified by liquid chromatography tandem mass spectrometry (LC-MS/MS) using a MASCOT database searching system. According to the results, there were 39 proteins associated with AAN. Focusing on ATP synthesis and anti-oxidation pathway, this study classified 12 of the 39 proteins into three categories: glycolysis, anti-oxidation, and ATP synthesis. Compared with AA group, most of the proteins related to glycolysis and ATP synthesis were significantly down-regulated in prednisolone group. On the other hand, proteins related to anti-oxidation were significantly up-regulated in prednisolone group comparing with AA group. These results indicate that prednisolone may slow the progression of AAN by decreasing glycolysis, ATP consumption and oxidative stress due to its known anti-inflammatory effect.
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Yeh, Hsin-I., and 葉欣怡. "Effects of ginseng and ginsenosides on aristolochic acid-induced nephropathy in inbred mice." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/82370543999051672394.
Full textAbstract:
碩士臺北醫學大學
藥學研究所
95
Aristolochic acid (AA) has been demonstrated to play a crucial role in Chinese herbs nephropathy. The purpose of this study was to evaluate the therapeutic effect of ginseng extrat (GE) and its active component, ginsenoside (GS), on AA-induced nephropathy. AA was dissolved in distilled water (3μg/ml) as drinking water to C3H/He mice (6 week-old male) for 56 days. The treatment groups were administered orally with GE (125, 250, 500 mg/kg) or ginsenosides (Rb1, Rd, Rg1 5 mg/kg) once daily for 14 days. The control group was administered with distilled water. The normal group was only administered with distilled water throughout the experiment. Urine protein (UP), urine N-acetyl-beta-D-glucosaminidase (NAG), blood urea nitrogen (BUN) and serum creatinine were determined to evaluate renal function. Renal tissues were served to histological examination (PAS stain and immunofluorescence). The antibodies, including TGF-β (transforming growth factor-β), MMP-9 (matrix metalloproteinase-9), HGF (hepatocyte growth factor), were chosen to recognize the specific antigens in injury sites. Compared with the control group, urine protein, NAG, BUN, serum creatinine and blood glucose were decreased in the GE 250 mg/kg, ginsenoside Rb1 and Rg1 treatment groups. In histological examination, we observed the alleviation in all treatment groups. According to the study, the effect of GE 250 is superior to the other GE-treated groups and ginsenoside Rg1 has the best effect among all the GS-treated groups.
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Huang, Yu Ping, and 黃昱斌. "Differential proteome analysis of aristolochic acid-induced nephropathy in mice with ginsenoside Rg1 treatment." Thesis, 2012. http://ndltd.ncl.edu.tw/handle/39390206849550267434.
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Chien, Chiung-hui, and 簡瓊惠. "Genotoxicity of aristolochic acid in human cells in vitro and in mice in vivo." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/36990088739756066392.
Full textAbstract:
碩士中國醫藥大學
中國醫學研究所
92
Aristolochic acid (AA), a probable human carcinogen found in Aristolochia fangchi, is thought to be a causative agent in Chinese Herb Nephropathy (CHN) patients developing urothelial carcinomas. To further understand the clastogenic and mutagenic effects of AA, the frequencies of micronuclei formation and gene mutations induced by AA were examined in vitro in human lymphoblastoid TK6 cells and in vivo in ICR mice. The exposure of TK6 cells to 25 to 200 g/ml AA for 4 hr produced a dose-dependent increase in cell killing and micronuclei formation. Treatment with 200 g/ml AA without S9 for 4 hr resulted in 73% survival and a 3-fold increase in the frequency of micronuclei (MN). In contrast, treatment with 200 g/ml AA with S9 for 4 hr resulted in 38.2% survival and a 8-fold increase in MN. A dose-dependent induction of hypoxanthine (guanine) phosphoribosyl transferase (hprt) and thymidine kinase (tk) mutations was observed after exposure to AA with various concentrations. Exposure to 200 g/ml AA for 4 hr either without or with S9 resulted in a 45~63-fold, 37~39-fold, and 2-fold increases in the mutant frequencies of hprt, tk-fast and tk-slow, respectively. Moreover, administration of AA by intraperitoneal injection and gavage (3 daily doses) at the dose of 40 to 60 mg/kg into ICR mice produced overt toxicity and caused statistically significant increase in the frequency of peripheral micronucleated reticulocytes 48 hr post exposure, about 30 and 9 times over control levels at a concentration of 60 mg/kg, respectively. These data showed that AA exhibits the clastogenic and mutagenic effects with overt cytotoxicity both in vitro and in vivo.
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Chen, Peng-Chi, and 陳鵬及. "Inflammation Involved COX-2 Induced by Aristolochic Acid Causes Heart Failure in Zebrafish Embryos." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/35276907986923668359.
Full textAbstract:
碩士國立臺灣海洋大學
生物科技研究所
94
“Chinese herbs nephropathy” (CHN) is a rapidly progressive nephropathy that was first described in 1992. Vanherweghem and his colleagues in Belgian found that women who had followed a particular weight-reducing regimen that included Chinese herbs developed CHN. Further plant biochemistry analyses revealed that the weight-reducing regimen contained the components of aristolochic acid (AA), which has been identified to be one of the causes to CHN. Robisch further showed that AA is also a mutagenic and carcinogenic agent. It has been reported that AA can cause tubular necrosis and interstitial renal fibrosis. However, the cellular or molecular mechanism of AA toxicity is largely unknown. We want to study the toxicity of AA using zebrafish as a model system. When treated zebrafish embryos with AA, we did not observe any defect in zebrafish kidney. Surprisingly, we found severe defects in the hearts. The heart phenotypes include 1) small and deformed heart, 2) less contractibility in atrium, 3) loss of contractibility in ventricle, 4) small heart lumen, and the other complications such as blood pool under the yolk, pericardial edema, termination of blood circulation and lethality. The heart phenotypes of AA-treated embryos are highly penetrant and reproducible. In order to distinguish whether AA affects the development or the physiology of the heart, we treated zebrafish embryos at different stages. We found that treatment on both young and old embryos causes similar phenotypes at later times. Also, we observed normal heart development in the AA-treated embryos up to at least 22 hours post fertilization when hearts begin to pump. These results suggest that AA may not directly affect heart development but its toxicity can be stored through embryogenesis and becomes affective after heart begins to function. To further understand the cellular defects underlie the contractibility phenotype, we tested whether AA affects the cardiac fiber structure by whole mount immunostaining with MF20 antibody which stains sarcomeres. AA treated hearts appear to have aggregation of cardiac fibers. By transmission electron microscopy, we observed dramatic defects in the cardiac fiber organization. The ventricular muscle fibers are largely missed and the remaining fibers appear loose and broken. Similarly, the atrial muscle fibers are disorganized and broken. However, we found that the expression of atrial muscle heavy chain (amhc) and cardiac muscle light chain 2 (cmlc2) is not significantly altered by AA treatment. Thus, we concluded that AA deleteriously affects the structure of cardiac fibers. By real-time PCR, we found that AA can induce the expression of IL-1β, SAA, COX-2 genes, which are involved in inflammatory response. To test whether inflammation is responsible for the AA toxicity, we treated zebrafish embryos with the anti-inflammation drug NS-398, which is a selective COX-2 inhibitor, together or after AA treatment. We found that NS-398 can attenuate the AA toxicity significantly. Furthermore, NS398 treatment reduces the expression level of COX-2 and SAA genes. Our studies using zebrafish embryos also provide insights into the toxicological mechanisms of AA. We first found that the EC50 is 5uM and AA toxicity is accumulative. For example: lower concentration or shorter treatment of AA will take longer time to cause the heart defects. With 10uM AA, we found that the minimal AA treatment time to generate toxicity is 4 hours. In summary, AA might trigger inflammatory response that causes specific destructive consequences to the cardiac muscle fibers. Our results with zebrafish embryos provide great details of cellular and molecular mechanisms of AA toxicity which is not yet possibly determined with other existing experimental systems.
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Ting-Yen and 朱鼎諺. "Induction of oxidative stress by Aristolochic acid in human renal proximal tubular epithelial cell line." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/72809582581373645923.
Full textAbstract:
碩士中山醫學大學
醫學研究所
97
Aristolochic acid (AA), a natural ingredient extracted from species Aristolochia, is composed of AAI and AAII. AAI is reported to be related to Chinese herb nephropathy (CHN), renal toxicity and carcinogenicity. To investigate whether AAI induce the generation of ROS and the toxicological relationship between AA and ROS, human proximal tubule epithelial cell line (HK-2) was chosen in our study. AAI (100 μM － 200 μM) treatment for 24 hours caused a marked decrease in the cell viability of HK-2. The ROS levels in HK-2 cells were also significantly induced by 200 μM AAI. The presence of antioxidant glutathione (GSH) effectively inhibited the ROS generation induced by AAI. Furthermore, in single cell gel electrophoresis assay, AAI at a concentration reached 50 μM resulted in the DNA damage in HK-2 nucleus. However, AAI-induced DNA damage was not suppressed by the co-existence of GSH. These results showed that AAI-induced DNA damage was not modulated by ROS generation. On the other hand, when human promyelocytic leukemia cell line (HL-60) was treated with AAI, the calcium concentration in HL-60 was significantly decreased in a dose-dependent manner. In view of cellular protective mechanism, with luciferase reporter assay we found that the Nrf2 binding elements were activated by AAI, which suggesting that AAI regulated the downstream antioxidant enzymes to against damage via activation of Nrf2 binding element. We further investigated he relationship between ROS and MAPKs signaling pathway. AAI induced the signals of phosphorylated ERK1/2 and p38 in HK-2 cells, but the presence of GSH could not effectively inhibit the phosphorylated signals induced by AAI. These results demonstrated that AAI-induced ROS is not a major upstream factor of ERK1/2 and p38 pathway. On the other hand, MEK1/2 inhibitor, U0126, dramatically decreased the AAI-induced phospho-ERK1/2 and ROS generation in HK-2 cells, indicating that AAI may mediate the ROS generation via MEK/ERK1/2 signaling pathway. Moreover, either p38 or ERK1/2 inhibitor reversed the DNA damage induced by AAI. These results implied that both ERK1/2 and p38 pathways play important roles in AAI-induced DNA damage.
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Sheau-Yun and 袁小雲. "Investigating the Action Mechanisms of Florin Extracts, Aristolochic acid and Tetrandrine on the Urinary System." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/92267867313600554856.
Full textAbstract:
博士中山醫學大學
醫學研究所
99
Objective: Calocedrus formosana (Florin) bark acetone/ethylacetate extracts are known to exert an antitumor effect on some human cancer cell lines, but the mechanism is yet to be defined. In this research, we investigated anti-bladder cancer cell activity in the leaf methanol extracts of Florin and the mechanism by which the extracts exert such activity. We also assessed the nephrotoxicity of aristolochic acid (AA) and tetrandrine (TET), the major bioactive components in the root extracts of Aristolochiae fangchi and Stephaniae tetrandra which have been used as diuretics and slimming agents. Materials and Methods: Florin leaf methanol extracts, and AA and TET were used for indentifying anti-bladder cancer activity and toxicity studies, respectively. Human bladder cancer cell lines, and normal renal cells (MDCK cells) and C3HeN mice were used in these studies. Methods included MTT test, flow cytometry, SDS-PAGE/Western blot, ELISA assay, immunofluorescence analyses, HE stain and TUNEL assay. Results: Florin extracts inhibited growth of bladder cancer cells by inducing G2/M phase arrest, attenuating β-tubulin polymerization and causing apoptosis as determined by MTT assay, flow cytometry, immunofluorescence staining, Western blot analysis of apoptotic proteins, respectively. In the nephrotoxicity assessment, although TET was more potent than AA in inhibiting MDCK cell growth in vitro as determined by apoptosis assay, mice treated with AA (10 mg/kg) by i.p. for 3 months showed higher nephrotoxicity, elevated blood urea nitrogen and increased renal tubular injuries when compared with those in mice treated with TET. Conclusions and suggestions: Florin extracts potently and specifically inhibit growth of human bladder cancer cells. AA but not TET exhibits a significant nephrotoxicity in mice under the experimental conditions. These suggest that Florin extracts may be an effective anti-bladder cancer agent and that the levels of AA in Chinese herbs should be an important concern for its nephrotoxicity.
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Cheng, Yu-Fan, and 鄭郁凡. "Effect of ginseng, ginsenoside Rg1 and prednisolone on aristolochic acid induced nephropathy in inbred mice." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/36757334484749717754.
Full textAbstract:
碩士臺北醫學大學
藥學研究所
96
Aristolochic acid (AA) has been demonstrated to play an important role in aristolochic acid nephropathy (AAN). The purpose of this study was to evaluate the therapeutic effect of ginseng extract (GE) or its active component ginsenoside Rg1 (GS), combined with prednisolone on AAN. AA was dissolved in distilled water as drinking water to C3H/HE mice (6 week-old male) for 56 days. The treatment groups in phase one were administered with GE 250mg/kg, prednisolone 2mg/kg, and both GE and prednisolone orally for 14 days. In phase two, under the same process, GE was replaced by GS. The control group was administered with distilled water and the normal group was only administered with distilled water throughout the experiment. Urine protein, urine N-acetyl-beta-D-glucosaminidase (NAG), blood urea nitrogen (BUN) and serum creatinine were determined to evaluate renal function. Renal tissues were served to histological examination (PAS stain and immunofluorescence). The antibodies, including TGF-β (transforming growth factor-β), MMP-9 (matrix metalloproteinase-9), HGF (hepatocyte growth factor), were chosen to recognize the specific antigens in injury sites.Compared with the control group, urine protein, NAG, BUN and serum creatinine were decreased at different level in all treatment groups. In the histological examination, we observed the alleviation in all treatment groups. The fluorescence dots of TGF-β were significantly decreased and MMP-9, HGF were significantly increased in experimental groups. Based on our result, both GE and GS combined with prednisolone has the superior effect among the concomitance groups. Our findings demonstrated that GE and GS are structure analogs of prednisolone and they have anti-inflammatory properties to slow down the fibrosis process and repair the renal injury.
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Ming-Chao and 劉銘超. "Aristolochic Acid Inhibits iNOS Gene Expression Induced by LPS or IFN-gamma in RAW 264.7 Macrophages." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/72786039528335604811.
Full textAbstract:
碩士中山醫學大學
生物醫學科學學系碩士班
98
Aristolochic Acid (AA), a group of natural compound widely found in Artistolochia species, is composed of AAI and AAII. AA I in the herbal medicine is found to be nephrotoxic and carcinogenic to human. To study the immunosuppressive ability of AAI, lipopolysaccharide (LPS) or Interferon gamma (IFN-γ) -stimulated RAW 264.7 macrophage cells were used as a model to examine the effects of AAI on the expression of the inducible nitric oxide synthase (iNOS) gene. When RAW 264.7 macrophages were treated with LPS (50 ng/ml) or IFN-γ (10ng/ml) for 18 h, the levels of nitric oxide (NO), iNOS protein and iNOS mRNA expression were all significantly increased. However, the presence of AAI (30-100μM) down-regulated the expression of LPS/IFN-γ-induced NO, iNOS protein and mRNA in a dose-dependent manner. To confirm whether AAI was able to decrease iNOS gene expression at the transcription level, we constructed a series of luciferase reporter plasmids which contained various promoter regions (-1588 to +121) of iNOS gene. AAI was found to inhibit the LPS/IFN-γ-induced iNOS expression by modulating the nuclear factor-κB (NF-κB) binding element located at nucleotides −86 to −76. The results of electrophoretic gel mobility shift assay (EMSA) also supported the inhibitory effect of AAI on the DNA binding activity of NF-κB. In addition, Western blotting demonstrated LPS-induced I-κB phosphorylation was significantly suppressed by AAI. Treatment of RAW 264.7 with AAI also down-regulated the LPS-induction of TNF-α and IL-6, two NF-κB regulated genes. Furthermore, the exposure of transient transfectant to AAI did not affect the luciferase activities of reporter construct that contained iNOS mRNA 3''-UTR, indicating that AAI does not inhibit iNOS gene expression at the post transcriptional level. Taken together, the data herein suggest that in activated RAW 264.7 macrophages, AAI regulates iNOS gene expression at the transcriptional level, and inhibition of NF-κB activation may be associated with the immunomodulatory effect of AAI.
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Cheng, Hui-Ni, and 鄭惠倪. "Therapeutic effects and molecular mechanisms of steroids on acute and chronic aristolochic acid nephropathy in mice." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/59785702840663995015.
Full textAbstract:
碩士中國醫藥大學
中醫學系
98
Aristolochic acid (AA), an established nephrotoxin and carcinogen, was known as the causal factor of Aristolochic acid Nephropathy (AAN). Clinical studies reveal that prednisolone therapy significantly slows the progression of renal failure in AAN patients. The transcription factor NF-κB has multiple crucial roles in the regulation of the immune/inflammatory responses, apoptosis, and carcinogenesis. In this study, we examined the correlation between the AA-induced nephritis and the activation of NF-κB, and the therapeutic effects and molecular mechanisms of steroids on acute and chronic AAN in mice Firstly, we used NF-kB/luciferase transgenic mice to construct an AA-induced nephritis model for noninvasive whole-body real-time imaging of NF-kb activity. NF-kB/luciferase transgenic mice were dosed by gavage with AA 4 mg/kg bw for 8 consecutive days, stopped for 2 days, and then gave prednisolone 1 mg/kg bw for 4 days. In vivo real time instantaneous imaging analysis showed that high levels of bioluminescence intensity were observed in AA-treated mouse kidney as compared with mock-treated mice, and prednisolone therapy significantly decreased the level of bioluminescence intensity. Microarray analysis of AA-treated mouse kidney and bladder tissues revealed that AA modulated the expression of genes involved in inflammation, DNA repair, carcinogenesis, extracellular matrix and cell adhesion. The down regulation by prednisolone of AA-induced NF-κB activation was confirmed by immunohistochemistry results demonstrated that AA profoundly increased the number of activated NF-κB p65-stained cells, while this increase was significantly decreased by prednisolone treatment. To investigate the protective effects of prednisolone on AA-induced inflammation and tumor formation, male FVB mice were dosed by gavage with AA 4 mg/kg bw for 14 consecutive days, then prednisolone (1 mg/kg bw and tapered off 1/2 every 2 weeks until 0.1 mg/kg bw) for 76 days, and followed by a life-time observation period. AA caused the tumor formation in kidney, stomach, liver and urinary bladder. However, after undergoing prednisolone treatment, the life-span of AA-exposed mice had significantly prolonged. Histopathological examination showed that prednisolone treatment significantly suppressed the formation of AA-induced renal and stomach carcinoma. Moreover, microarray analysis of renal and bladder tissues exposed to AA with or without prednisolone showed that several signal pathways involved in inflammation and carcinogenesis were significantly regulated. Results from immunohistochemistry demonstrated that the number of NF-κB stained cells in AA-treated groups as compared to Mock groups while this increase was significantly decreased by prednisolone treatment. Our results revealed that the anti-inflammatory effect of prednisolone may partly account for its beneficial effect on the life-span, its suppression on AA-induced inflammation and possibly the tumor formation in AA-treated mice, and those effects are may be through the down-regulation of NF-κB. Glycyrrhizinic acid (Glycyrrhizin), a steroid-like glycoside obtained from Glycyrrhiza glabra, possesses anti-inflammatory activities. In this pilot study, we examined the effects of glycyrrhizinic acid on acute AAN. Male FVB mice were dosed by gavage with AAⅠ (2.5 mg/kg bw) for 8 consecutive days, and stopped for 2 days, then gave glycyrrhizinic acid (25 mg/kg bw) for 7 days, or FVB mice were dosed by gavage with AAⅠ (2.5 mg/kg bw), 5 minutes later, gave glycyrrhizinic acid (25 mg/kg bw) for 17 consecutive days. Since there is no significant increase of urinary total protein/creatinine ratio, plasma creatinine and BUN level after AA exposure, further experiments are needed to examine the effects of glycyrrhizinic acid on acute AAN.
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Chiang, Xin-Yu, and 蔣欣妤. "The effect of aristolochic acid and BLCAP gene on Smad signaling pathway in human kidney cell line." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/p7puf8.
Full textAbstract:
碩士國立臺灣大學
毒理學研究所
106
Aristolochic acid (AA), a group of natural compound widely found in Aritolochia family of plants, is composed of AAI and AAII. Aristolochic acid nephropathy (AAN) is a rapidly progressive tubulointerstitial fibrosis associated with the intake of AA. Bladder cancer associated protein (BLCAP) is considered to be a potential tumor suppressor gene in numerous cancers. However, the biological function and regulation of BLCAP gene remains unclear. The aims of this study are focused on whether AAI-induced Gremlin upregulation is mediated by BLCAP gene and on the effects of AAI and BLCAP gene on the Smad signaling pathway in human embryonic kidney 293 (HEK293) cell line. Exposure of HEK293 cells to AAI for 24 h significantly increased the expression of Gremlin mRNA. This increase was in parallel with a down-regulation in BMP-7 mRNA and phosphorylated Smad1/5/8 protein levels. Besides, AAI also reduced the level of BLCAP transcript. To examine whether AAI regulated the expression of Gremlin through BLCAP gene, HEK293 cells were infected with lentivirus harboring pLKO.1-shBLCAP (shBLCAP/HEK293) to decrease the level of BLCAP mRNA. Downregulation of BLCAP transcript in shBLCAP/HEK293 cells did not affect the levels of Gremlin and BMP-7 mRNA. We found that Smad1/5/8 phosphorylation and its downstream Id2 mRNA were inhibited in shBLCAP/HEK293 cells due to a lower level of BMP type-II receptor (BMPR2) mRNA. On the other hand, knocking down BLCAP promoted the cell growth of shBLCAP/HEK293 cells, and also enhanced TGF-β1 mRNA signals, Smad2/3 phosphorylation and the expression of downstream targets, including PAI-1, CTGF, and microRNA-21 (miR-21). Transfection of miR-21 inhibitor into shBLCAP/HEK293 cells could recover the cellular proliferation induced by BLCAP deficiency and also restore the decreased signal of BMPR2 mRNA. Taken together, AAI induced the gremlin expression and its downstream BMP-7/Smad1/5/8 signaling in HEK293 cells without the involvement of BLCAP. Furthermore, BLCAP gene not only regulated Smad1/5/8 pathway by modulating BMPR2 expression, but also negatively affect TGF-β1/Smad2/3 signaling.
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Tseng, I.-Hua, and 曾一華. "Detection of Chymase Activity Using Gold Nanoparticles Peptide Probe and Studying the Role of Chymase in Nephropathy Induced by Aristolochic Acids." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/vga568.
Full textAbstract:
碩士國立交通大學
生物科技學系
103
Gold nanoparticles (AuNPs) are the most popular nanomaterials due to its unique optical properties and feasibility of surface modifying with biomolecules. AuNPs with the characteristics of surface plasmon resonance (SPR) and fluorescence resonance energy transfer (FRET) have the ability of quenching fluorescence with distant dependece. Based on this, we have established a sensitive and efficient biosensing method by modifying peptide-probe onto the AuNPs to detect enzyme activity in this study. The biosensing method is designed for chymase activity detection and applied in renal disease diagnosis. Aristolochic acid (AA) is an extract from Chinese herbs which causes progressive interstitial nephritis. In kidney, angiotensin converting enzyme (ACE) is the primary enzyme which converts angiotensin I (Ang I) to angiotensin II (Ang II) in renin-angiotensin system (RAS). However, in some renal diseases, several evidences have showed that chymase may play a role to convert Ang I into Ang II independent to ACE. The aim of this research is to investigate whether chymase play the crucial role in AA-induced nephropathy. In this study, 13-nm AuNPs were used to establish the AuNPs-based fluorescence peptide probe (named AuNPs-peptide probe) for chymase detection. The peptide sequence is FITC-Acp-DRVYIHPFHLDDDDDC which contains a fluorophore at the C-terminal end, enzyme (chymase) substrate (DRVYIHPFHL), a spacer (DDDDD) and a cysteine (C) used for conjugating to AuNPs surface. A red shift of O.D. was observed from 520 nm to 525 nm after the 13-nm AuNPs modified with the peptide into be AuNPs-peptide probe. When the enzyme catalyzed the substrate sequence, the fluorophore drifted away from AuNPs and the fluorescence emitting signal can be excited at 485 nm and detected at 515 nm. In the animal experiments, animal model of nephropathy was performed in wild type (C57BL/6, WT) and ACE2 knockout (ACE2 KO) mice. The mice were treated with aristolochic acid I (AAI), which is the major and toxic constituent of AA, via intraperitoneal (ip) injection and the accumulated AAI dosages are 30 and 100 mg/kg of body weight. The animals were sacrificed after another 2 or 4 weeks for nephropathy development, and blood and kidney were sampled for the further biochemical, pathologic and molecular assays. The results show, detection time of the AuNPs-peptide probe used for the activity detection of chymase only needed 15 min and a linear correlation from 10 to 120 ng/mL of chymase is acquired. Moreover, detection limit of the AuNPs-peptide probe is 5 ng/mL. The chymase reaction would be obviously inhibited by adding the chymase antagonist (chymostatin). The AuNPs-peptide probe was also to detect high concentration of trypsin and chymotrypsin, but no emitting fluorescence intensity was detected. According to the results, sensitivity and specificity of the AuNPs-peptide probe using for chymase detection have been approved. The mice after the treatment of AAI, decreased bodyweight and enhanced serum creatinine as well as increased blood urea nitrogen (BUN) were observed. In the renal tissue sections, high amount of inflammatory cells were found by hematoxylin and eosin (H&;E) stain, and increased blue areas of fibrosis in renal interstitial tissue were observed by Masson’s trichrome stain. The biochemical and pathologic characterizations in ACE2 KO mice were more severe than those in WT mice. In WT mice, increased MMP-2 and decreased MMP-9 activity were found in the AAI-treated mice compared with those untreated controls. More, an significantly increase in chymase activity was also found treated with AAI. However, in the AAI-treated kidney tissue, both ACE and ACE2 activity did not show significant changes, indicating the chymase may play the important role in the nephropathy progress. In ACE2 KO mice, the changes of MMP-2, MMP-9 and ACE activity were similar with those in WT mice. However, the increased chymase activity in ACE2 KO mice was more significant compared with that in WT mice. According to the results, sensitivity and specificity of the AuNPs-peptide probe using for chymase detection have been approved and it is effective to detect chymase activity in kidney tissue. In the AAI-induced nephropathy, ACE and ACE2 activity in kidney tissue did not show significant change, but chymase did, showing that the chymase may play a crucial role in the AAI-induced kidney damage. Moreover, it was also observed that the deficiency of ACE2 would accelerate the disease development of AAI-induced nephropathy. These results may help physicians and researchers to know more information about the role of RAS in kidney disease and can be applied in developing new drug targets for nephropathy.
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Wen-Chi, Yang, and 楊文淇. "Establishment of the aristolochic acid nephropathy in inbred mice and the effect of Bupleuri Radix on the nephritis." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/34463192650084569620.
Full textAbstract:
碩士臺北醫學大學
藥學系
93
Nephrotoxicity induced by weight-control Chinese medicine was reported frequently in recent years. One of the factors is mistaken the herb that contains aristolochic acid (AA). Aristolochic acid nephropathy (AAN) is a unique type of nephropathy, which was characterized by an extensive interstitial fibrosis with rare cell infiltration and tubular atrophy. We tried to establish a chronic interstitial fibrosis model of AAN in inbred mice and investigated the impact of Bupleuri Radix, prednisolone on AAN. Aristolochic acid (AA) dissolved in distilled water (3μg/ml) were administered orally to C3H/He mice (6 weeks, male) as drinking water ad libitum for 4 weeks, control group were treated with distilled water. In the second experiment, they were administered orally with Bupleuri Radix extract (BR) (150, 300, 450 mg/kg), saikosaponin A, C, D, prednisolone (P), BR 300 mg/kg combine with P once a day for 14 days. The control group was treated with distilled water. Urinary protein, urinary N-acetyl-beta-D-glucosaminidase (NAG), blood urea nitrogen (BUN), blood glucose were determined. Renal tissues were served to histological examination (PAS stain and immunofluorescence staining). The antibodies, including murine F4/80 macrophage, TGF-β (transforming growth factor-β), MMP-9 (matrix metalloproteinase-9) was chosen to recognize the specific antigens, which deposited in injury sites. All animals given AA developed elevation of urinary protein, NAG, BUN, blood glucose. The histological examination was observed the typical AAN: tubular atrophy, interstitial infiltration and fibrosis. In the immunofluorescence stain assay, macrophage, TGF-β, MMP-9 were localized in the renal tissue. After discontinuation of AA, renal function was not improved. The amount of urinary protein, NAG, BUN, blood sugar were decreased in the BR 300 mg/kg, saikosaponin A, D, P, BR combine with P-treated groups compared with the control group. The histological examination was observed the alleviation in all treatment groups. The staining areas of macrophage, MMP-9 in the interstitium were significantly increased, and TGF-β was decreased among all groups. Our result demonstrate the therapeutics effect of the treatment as following: BR combine with P have more prior effect than P, BR 300 mg/kg, saikosaponin D, saikosaponin A. This present study offers a new therapeutic trend of utilizing the crude drug preparation with medicine.
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Chou, Yung-Chen, and 周泳臣. "INVESTIGATION OF THE EFFECTS OF ARISTOLOCHIC ACID ON MONOCYTIC MATRIX METALLOPROTEINASE-9 ACTIVATION AND NON-SPECIFIC IMMUNE FUNCTION." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/54456307752248704440.
Full textAbstract:
碩士臺北醫學大學
藥理學研究所
93
Many evidence indicated that rheumatoid arthritis (RA) is a disease including immune and inflammatory systems which are linked to the destruction of cartilage and bone. Monocyte/macrophages, lymphocytes and fibroblasts are found in highly increased numbers in the synovial membrane in RA. Leukocytes and synovial fibroblast overproduce pro-inflammatory cytokines, mainly tumor necrosis factor-alpha (TNF-), interleukin (IL)-1 and IL-6, which lead to chronic inflammation. These cytokines activate a variety of gene expression, including genes coding for various cytokines and matrix metalloproteinases (MMPs) involved in tissue degradation. On the other way, MMPs also play an important role in tumor invasion. The invasive properties of tumor cells, owing to their ability to degrade all major protein components of the extracellular matrix (ECM) and basement membranes. They also participate in early steps of tumor evolution, including stimulation of cell proliferation and modulation of angiogenesis. We found that aristolochic acids (AsA) extracted from herbal medicines (such as Aristolochia spp) showed obviously inhibitory effect on MMPs activation. In this study, we found that AsA was shown to inhibit the TNF--induced MMP-9 activation and expression in human monocyte THP-1 cells by zymography method and Western Blot (IC50= 6.41 ± 0.45 M). We also found the different ratio (I : II = 55:42 and 29:66) of aristolochic acids I and II with similar effect on inhibition of MMP-9 activation. We also used indomethacin to exclude the inhibitory action of AsA involved in prostaglandin pathway. In the transcription level, AsA suppressed the TNF--induced MMP-9 mRNA expression by using RT-PCR. AsA also inhibit the TNF--induced IB- degradation. In the nuclear aspect, we also found that AsA inhibited TNF--induced NF-B activation (by EMSA method) and translocation. Interestingly, AsA was shown to concentration-dependently inhibit the concanavalin A (Con A) and lipopolysaccharide (LPS) induced proliferation of mouse splenocytes. We also compare the inhibitory effect of AsA with dehydroepiandrosterone (DHEA), and the inhibitory effect is similar. By the way, the cytokines (IL-2 and interferon-) released from the ConA-induced lymphocyte were concentration-dependently affected. However, the inhibitory mechanisms of AsA on lymphocyte need further investigated. It will be interesting to study further the anti-inflammatory activities of this nature compound on RA-related synovial injuries in vivo.
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Liang, Meng-Chieh, and 梁孟傑. "Analysis of aristolochic acid I and its application in urine by surface-enhanced Raman scattering and photoluminescence spectra." Thesis, 2017. http://ndltd.ncl.edu.tw/handle/33hznn.
Full textAbstract:
碩士國立中山大學
化學系研究所
105
Controversy over the health effects of Chinese herbal medicines containing aristolochic acid remains unresolved. Multiple cases of renal fibrosis and renal failure have been reported in patients taking Chinese medicines. In this study, a novel biochemical method for the detection of Aristolochic acid I (AAI) is proposed. We used silver nanoparticles (Ag NPs) for surface enhanced Raman scattering (SERS) and fluorescent Epoxy nanodots (Epoxy NDs), as substrates for detection of AAI by SERS and Photoluminescence (PL) spectra respectively. For SERS experiments, two types of complexes were formed. In the first, AAI was bound to 3-Aminopropylsilatrane (APS) coated Ag NPs. Silver ions were reduced to form spherical (5.48 ± 0.14 nm) Ag NPs, then an APS layer was formed, creating a large aggregate (4.78 ± 0.09 μm) and finally AAI was coated on the APS to form AAI/APS@Ag NPs. Here, APS was not only used as a reducing agent, but also a coating agent. In the second method, AAI was directly bound to the Ag NPs and then an outer coating of APS was applied to form APS/AAI@Ag NPs. The proposed mechanism for Ag NPs synthesis was confirmed by X-ray photoelectron spectra (XPS) and Fourier transform infrared (FTIR). The reduction of Ag NPs was confirmed by XPS and Energy-dispersive X-ray spectra (EDS). We investigated the best reducting conditions of Ag NPs by SERS, and measured the enhancement factor of up to 10⁵. The limit of detection (LOD) of SERS is 10 μM. For PL experiment, the modified nitroreducted AAI underwent aminolysis reaction on the surface of Epoxy NDs, which served to enhance the fluorescence intensity. The modification of Epoxy NDs was confirmed by XPS and FTIR. The LOD of PL is 2 μM. In the body, AAI is metabolized to form Aristololactam Ia (ALIa). The two biochemical methods demonstrated in this study for detecting ALIa are easily prepared, economical, and don’t require complicated pretreatment of the samples. These may provide a superior alternative to existing detection methods for ALIa in clinical studies.
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Chen, Po-Hsun, and 陳柏勳. "Tackling the Risk of Herbs under the Globalization of Chinese Herbs: Aristolochic Acid Controversy and Response in Taiwan." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/7m99rf.
Full textAbstract:
碩士國立陽明大學
科技與社會研究所
106
In the late 20th century, the Aristolochic Acid controversies not only changed the concept of herbs in the international but transformed the regulation of herbs in Taiwan. By analyzing the Aristolochic Acid controversy in Taiwan, this thesis will investigate the following questions. How and why were the herbs considered toxic and risky? How did Taiwan government making policies by negotiating between the traditional Chinese medicine and the modern science? How did the multiple actors build the new concepts and new regulations of herbs under the globalization? At last, how did the Aristolochic Acid controversy structurally transform the herb regulation? The Aristolochic Acids are in Aristolochiaceae herbs, which are known for the renal toxicity and carcinogen. It was the Aristolochic Acids that cause the Chinese herb nephropathy in Belgium after the investigation in 1993. Aftermath, there were more and more poisoned cases around the Europe, America, and Asia. Thus, many countries set the bans on the preparations which contained the Aristolochic Acids. The toxicology medicine physician gave a speech about the renal toxicity of aristolochic acid in Chinese Herbal Toxicity Congress in Taipei in 2003. Also, a traditional Chinese medicine doctor made an appearance and claimed that he was the victim of Aristolochic acids. It aroused a great controversy between the modern medicine and the traditional Chinese medicine for prohibiting Aristolochiaceae herbs or not. This controversy, on the one hand, highlighted that it is unable to frame the toxicity and the efficacy of herbs by one type of knowledge. On the other hand, it also raised the public alert to the toxicity of herb. The regulation for the toxicity of herb became the essential issue to Taiwan government for expanding the global market of herb. To adjust the impact of WHO Traditional Medicine Strategy in 2002 and to join WTO in 2005, the Chinese medicine could not uphold the traditional regime to regulate the herb. However, they were not willing to accept the framework of biomedicine. The Chinese medicine integrated the modern technology and the concept of materia medica to build the new knowledge and the new regulations for herbs. There was also the new idea for herbal toxicity. The big data researches, such as the epidemiology and the National Health Insurance Research Databases, turned into the important scientific means to comprehend the efficacy, safety, and risk of herbs. As a result, the new knowledge and the new regulation model of herbs were taken shape. The government took this knowledge and system not only to quell the public doubts but to act as the quality guarantee for herbal preparations in the global market. Even though, there was some room for improvement in this new herbal safety system. In sum, after the Aristolochic Acid controversy, the regulation of herb turned into the regime of risk in Taiwan. Different from following the model of USFDA to prohibit the Aristolochic-Acid contained herbs, the Chinese medicine decided to establish the safety environment and negotiated the risk of herbs. Under the push-pull factors of herbal risk and global market, the Chinese medicine heterogeneously integrated knowledge, system, and actors to construct the safety of herbs by decreasing the risk of herbs. Furthermore, the Chinese medicine made the modern conventions to the quality of herbs in Taiwan.
49
Wang, Hsin-Yu, and 王欣玉. "Investigation of the Inhibitory Mechanisms of N-hydroxycinnamoylphenalkylamides Analogues and Aristolochic Acid on MCP-1-Activated Monocytic cells." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/03473602361414176406.
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碩士臺北醫學大學
醫學科學研究所
97
In the human innate immune system, the monocytes are the key cellular elements and serve as a critical line of defense. Inflammatory monocytes respond rapidly to microbial stimuli by secreting cytokines and antimicrobial factors, express the CCR2 chemokine receptor, and traffic to sites of microbial infection in response to monocyte chemoattractnat protein (MCP-1) secretion. On the other hand, the monocytes also can differentiate to macrophages and proceed a more powerful, long-term immunoreaction. When normal regulatory mechanisms fail, the monocyte is also responsible for immunologically induced many physiological and pathological changes, like atherosclerosis, rheumatoid arthritis (RA) and cancer. In in vitro invasion and migration assay, we found that N-hydroxycinnamoylphenalkylamides analogues (EK5 and EK8) and Aristolochic acid (AsA) showed obviously inhibitory effect on MCP-1-induced monocyte invasion and migration. The MEK inhibitor PD98059 and PI3K inhibitor LY294002 also inhibited MCP-1-induced monocyte invasion, so monocyte invasion might be regulated by ERK1/2 and PI3K signaling pathway. We also found AsA and EK5 might inhibit MCP-1-induced monocyte tethering (rolling) to firm adhesion via integrin activation and MMP-9 activity by affecting Akt signaling pathway. And AsA might inhibit MCP-1-induced monocyte ??1 integrin activation and actin rearrangement by affecting ERK1/2 signaling pathway. But EK8 had no effect on Akt and ERK1/2 signaling.
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Lung, Tsai Jia, and 蔡佳龍. "Deternmination of Aristolochic acid , the trace toxin in Chinese herb Xixin（Asarum spices）by liquid chromatography-mass spectrometry." Thesis, 2002. http://ndltd.ncl.edu.tw/handle/55990606417128653244.
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