Dissertationen zum Thema „KMIP“

Tyrimo probleminis klausimas: ar laisvalaikiu sportuojančių moterų mityba yra sveikesnė nei nesportuojančių ir ar yra ryšys tarp laisvalaikiu sportuojančių moterų kūno kompozicijos ir maisto raciono. Tyrimo objektas – laisvalaikiu sportuojančių ir nesportuojančių moterų kai kurie kūno kompozicijos rodikliai ir mitybos ypatumai. Tiriamojo darbo tikslas buvo nustatyti laisvalaikiu sportuojančių ir nesportuojančių moterų mitybos ypatumus bei sąsają su riebaline kūno mase. Iškėlėme tokius darbo uždavinius: 1. Įvertinti tiriamųjų kai kuriuos kūno kompozicijos rodiklius (riebalines odos raukšles, KMI, riebalinę kūno masę) bei palyginti juos tarp laisvalaikiu sportuojančių ir nesportuojančių moterų. 2. Įvertinti laisvalaikiu sportuojančių ir nesportuojančių moterų mitybos ypatumus bei juos palyginti. 3. Įvertinti laisvalaikiu sportuojančių moterų mitybos sąsają su riebaline kūno mase. Tyrimo metodai – literatūros apžvalga, antropometriniai matavimai, kūno riebalinės masės procentinis apskaičiavimas, anketinė apklausa, mitybos raciono apklausa, analizė ir matematinė statistika. Tyrimo organizavimas: mūsų tyrimas buvo pradėtas vykdyti 2011 metų gegužės mėnesį, o baigtas – 2012 sausio mėnesį. Tyrimo eigos metu vyko tiriamųjų pasirinkimas, supažindinimas su tyrimo tikslais, metodais. Šiame tyrime dalyvavo dvi tiriamųjų moterų grupės – sportuojančios ir nesportuojančios. Laisvalaikiu sportuojančių moterų grupė buvo iš n=50 tiriamųjų, kurių amžius 25±5 metai. Nesportuojančių... [toliau žr. visą tekstą]
Research problem question: whether leisure exercising women’s nutrition is healthier than unexercising women’s, and whether there is a connection between leisure exercising women’s body composition and diet. The object of research – some of leisure exercising women’s and not exercising women’s body composition details and their nutrition habits. The aim of the study was to determine the connection between the leisure exercising and not exercising women's nutrition peculiarities and their body fat mass. We set the following tasks: 1. To evaluate the research of some indicators of body composition (fat skin folds, BMI, fat body mass) and to compare them among recreational athletes and untrained women. 2. To rate leisure exercising women's and not exercising women's feeding habits and compare them. 3. To rate connection between leisure exercising women’s nutrition and body fat mass. Research methods - review of literature, anthropometric measurements, body fat mass percentage calculation, a questionnaire of nutrition, analysis and mathematical statistics. Research organization: our investigation was launched in 2011 May and completed - 2012 January. During the study course we selected objects, introduced them to research methods. There were two groups of women - leisure exercising (50) aged 25 ± 5 years and not exercising (30) age 24 ± 5 years. After this study, we made the following conclusions: 1. In both groups, leisure exercising and unexercising women, body mass... [to full text]

This thesis deals with the study of soils freezing in terms of phase change of water contained in the soil strata on green roofs. The aim of this work is to verify the effect of phase transformation of water on the course of temperature oscillation. First described the basic characteristics of soils generally, and subsequently described processes occurring during phase transformation of water in the soil and has been carried out experimental verification of the effect of moisture in the soil on the course of temperature oscillation. In the overall evaluation of the work, an analysis of the effect of phase change water in soil on the course of temperature oscillation and the resulting conclusion of work.

The goal of this thesis was to develop, deploy and evaluate a lightweight portable intrusion detection system (LPIDS) over wireless networks. The LPIDS was developed by adopting two different string matching algorithms: Aho-Corasick algorithm and Knuth–Morris–Pratt algorithm (KMP). The LPIDS was implemented and tested on the hardware platforms Wi-Fi Pineapple and Raspberry Pi. To evaluate and test the LPIDS as well as the algorithms, performance metrics such as throughput, response time and power consumption are considered. The experimental results reveal that Aho-Corasick performed better than KMP throughout the majority of the process, but KMP was typically faster in the beginning with fewer rules. Similarly, Raspberry Pi shows remarkably higher performance than Wi-Fi Pineapple in all of the measurements. Moreover, we compared the throughput between LPIDS and Snort. It was concluded that the throughput was significantly higher for LPIDS when most of the rules do not include content parameters. This thesis concludes that due to computational complexity and slow hardware processing capabilities of Wi-Fi Pineapple, it could not become suitable IDS in the presence of different pattern matching strategies. Finally, we propose a modification of Snort to increase the throughput of the system.

Programa de Pós-Graduação em Ciências Biológicas. Núcleo de Pesquisas em Ciências Biológicas, Pró-Reitoria de Pesquisa e Pós Graduação, Universidade Federal de Ouro Preto.
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O presente estudo avaliou a toxicidade/inocuidade, imunogenicidade e eficácia/potência dos protótipos vacinas KMP-11 e LBSap, de forma comparativa com as vacinas comerciais Leishmune® e Leish-Tec®, em um ensaio clínico vacinal de fase I e II. Para isso, trinta e cinco cães foram classificados em cinco grupos (sete cães por grupo): i) grupo controle receberam 1 mL de solução salina estéril 0,9%; ii) grupo Leish-Tec receberam a vacina comercial Leish-Tec®; iii) Grupo Leishmune receberam a vacina comercial Leishmune®; (iv) grupo LBSap recebeu 600 μg de proteína de promastigotas de L. braziliensis e 1 mg do adjuvante saponina; (v) grupo KMP-11 receberam 100 μg do antígeno recombinante KMP-11 associado a 1 mg do adjuvante saponina. Uma análise detalhada e comparativa da inocuidade/toxicidade vacinal foi realizada através do quadro clínico, bioquímico e hematológico entre as doses de imunização e após o término das três doses vacinais. Embora em alguns cães dos grupos KMP-11, LBSap e Leishmune tenham apresentado alterações no local dos inóculos vacinais, relacionados ao adjuvante saponina, como: nódulos, edema leve, dor local, estes foram transientes e desapareceram após setenta duas horas da vacinação. Nossos resultados de inocuidade indicam que as alterações adversas provocadas pelas imunizações são toleráveis, tendo em vista a importância e funcionalidade que uma vacina canina eficaz promoveria no controle da doença. Nossos resultados da imunogenicidade vacinal demonstram um aumento da população circulante de linfócitos T CD8+ ao final do protocolo vacinal (T1) nos grupos LBSap e Leish-Tec, além de seis meses após o desafio experimental (T2) no grupo LBSap. Também foi observado em T1 aumento de linfócitos B (grupo Leishmune) e de monócitos CD14+ (grupos LBSap, Leishmune e KMP-11), que justificam e reforçam o potencial imunoprofilático destas vacinas. Nas análises in vitro foi observado aumento na atividade linfoproliferativa nos grupos LBSap e Leishmune, sendo no grupo LBSap um aumento antígeno-específico de linfócitos TCD4+ e CD8+. Uma segunda abordagem das análises in vitro buscou avaliar o percentual de células T CD4+ e CD8+ antígeno-específicas produtoras de IFN- e IL-4, onde foi observado no grupo LBSap aumento de ambas as subpopulações produtoras de IFN-, sendo também evidenciado um aumento de T CD8+ produtores de IFN- no grupo Leish-Tec. Nosso resultados de imunogenicidade reforçam a hipótese que o processo vacinal, principalmente, com a vacina LBSap levam a geração de uma resposta imune protetora contra o agente etiológico da LVC compatível com o controle do parasito de L. infantum. Nossos resultados da reatividade sorológicos demonstraram que o TR DPP® foi capaz de distinguir os cães doentes dos vacinados. Após o desafio experimental pela via endovenosa os cães foram acompanhados por 6 meses e foram evidenciadas alterações clínicas sugestivas da infecção por Leishmania, entretanto, na maioria dos cães foi observado uma infecção assintomática. Na avaliação parasitológica da medula óssea foi possível isolar o parasito (mielocultura) bem como quantificar o DNA (qPCR) de Leishmania em todos os grupos, sendo observado redução considerável da carga parasitária da medula óssea em todas as vacinas testadas em relação ao grupo Controle. Entretanto, no grupo de cães imunizados com LBSap esta redução foi mais evidente, chegando a ser 47 vezes menor. Com base no que foi exposto, a vacina LBSap seria a mais indicada para prosseguimento em ensaio clínico vacinal de fase III, em relação a vacina KMP-11. ____________________________________________________________________________________________
ABSTRACT: This study evaluated the toxicity/safety, immunogenicity and efficacy/potency of vaccines prototypes KMP-11 and LBSap, in comparison to the commercial vaccines Leishmune® and Leish-Tec®, in a vaccine clinical trial phase I and II. For this, thirty-five dogs were classified into five groups (seven dogs per group): i) control group received 1 mL of sterile 0.9% saline solution; ii) Leish-Tec group received the Leish-Tec® commercial vaccine; iii) Leishmune group received the Leishmune® commercial vaccine; (iv) LBSap groups received 600 μg of L. braziliensis promastigotes protein and 1 mg of saponin adjuvant; (v) KMP-11 group (n=7) received 100 μg recombinant KMP-11 antigen associated to 1 mg saponin adjuvant. A detailed and comparative analysis of safety/toxicity vaccination was performed by clinical, biochemical and hematological parameters between dose and immunization after the end of the three vaccine doses. Although some groups of dogs KMP-11, LBSap and Leishmune have presented changes at the site of vaccination inoculum, related to saponin adjuvant, such as nodules, mild edema, and local pain, these were transient and disappeared seventy two hours after vaccination. Our results indicate that the safety of adverse changes caused by immunizations is tolerable in view of the effective canine vaccine importance and functionality that it promotes in the disease control. Our results of the immunogenicity vaccine demonstrate increase in circulating population of T CD8+ lymphocytes in the end of the immunization protocol (T1) in groups LBSap and Leish-Tec, as long as six months after experimental challenge (T2) in LBSap group. It was also observed in T1, an increase of B lymphocytes (Leishmune group) and monocytes CD14+ (LBSap, Leishmune and KMP-11 groups), that justify and reinforce the potential immunoprophylactic of these vaccines. In the in vitro analyzes an increase in lymphoproliferative activity in groups LBSap and Leishmune was observed, occurring in LBSap group an antigen-specific increase of CD4+ and CD8+ T-lymphocytes. A second approach of in vitro assays aimed to evaluating the percentage of antigen-specific CD4+ and CD8+ lymphocytes producers of IFN- e IL-4, where an increase in both IFN- producing subpopulations in the group LBSap was observed, and it also showed an increase in IFN- producers in CD8+ lymphocytes in the Leish-Tec group. Our immunogenicity results support the hypothesis of the vaccine process, especially with the LBSap vaccine generating a protective immune response against the causative agent of CVL compatible with L. infantum parasite control. Our results of serological reactivity demonstrate that TR DPP® was able to distinguish among diseased and vaccinated dogs. After experimental challenge by intravenous route the dogs were followed for 6 months and clinical changes suggestive of Leishmania infection were observed, however in the majority of dogs asymptomatic infection was observed. In the parasitological evaluation of bone marrow it was possible to isolate the parasite (myeloculture) and quantify the DNA (qPCR) of Leishmania in all groups, significant reduction in parasite burden in the bone marrow was observed in all vaccines tested compared to control group. However, in the group of dogs immunized with LBSap this reduction was more evident, being 47 times smaller. Based on the foregoing, the LBSap vaccine would be the most suitable for further research in phase III clinical trial vaccine in relation to KMP-11 vaccine.

There is no doubts that proper and healthy nutrition helps to prevent a number of chronic non-communicable diseases and is one of the key determinants of good health and life quality. The aim of the study is to evaluate the actual nutrition and nutrition habits of Lithuanian adult population. A random sample of 3,000 Lithuanian residents aged from 19 to 65, representing Lithuanian adult population, was set for this study. Nutrition habits were investigated using nutrition questionnaire, food consumption was investigated using 24 hours recall methodology and special Atlas of Foodstuffs and Dishes portion sizes. Presented study is the first case, where the complexed data about the actual nutrition, nutrition habits and BMI of Lithuanian adult population was analyzed and comprehensively evaluated. According to socio-demographic determinants it was examined and evaluated consumption of separate foodstuffs as well as daily intake of nutrients (proteins, fats (including saturated and unsaturated fatty acids), carbohydrates (including sugars), dietary fibers, vitamins, minerals, etc.), and their consistency within the recommendations. There were established trends of changes in Lithuanian adult population nutrition and BMI, as well as links between respondents’ attitude towards some aspects of the nutrition (nutrition impact on health, consumption of fruits and vegetables, consumption of products containing more saturated fatty acids, heavy salt consumption, iodine salt consumption... [to full text]
Tinkama ir sveika mityba padeda išvengti daugelio lėtinių neinfekcinių ligų ir yra vienas iš svarbiausių veiksnių, lemiančių žmonių sveikatą ir gyvenimo kokybę. Todėl šio darbo tikslas buvo įvertinti suaugusių Lietuvos gyventojų faktišką mitybą bei mitybos įpročius. Buvo sudaryta atsitiktinė 3000 Lietuvos suaugusių (19–65 m. amžiaus) gyventojų imtis; mitybos įpročiai tirti, naudojant apklausos anketą; faktiškos mitybos tyrimai atlikti pagal standartinę 24 valandų apklausos metodiką, panaudojant specialiai šiam tikslui parengtą Maisto produktų ir patiekalų porcijų nuotraukų atlasą. Disertaciniame darbe pirmą kartą išanalizuoti ir kompleksiškai įvertinti duomenys apie suaugusių Lietuvos gyventojų faktišką mitybą, mitybos įpročius, gyventojų KMI; išnagrinėtas ir įvertintas pagal sociodemografines determinantes atskirų maisto produktų suvartojimas bei su jais gaunami maistinių medžiagų (baltymų, riebalų, tarp jų sočiųjų ir nesočiųjų RR; angliavandenių, tarp jų cukrų, skaidulinių medžiagų, taip pat vitaminų, mineralinių medžiagų ir kt.) kiekiai per parą bei jų atitikimas Rekomenduojamoms paros normoms. Taip pat buvo įvertintos Lietuvos suaugusių gyventojų mitybos ir kūno masės indekso pokyčių tendencijos bei nustatytos mitybos ir mitybos įpročių sąsajos su respondentų požiūriu į tam tikrus mitybos aspektus (mitybos įtaką sveikatai, daržovių ir vaisių vartojimą, produktų, turinčių daugiau sočiųjų riebalų rūgščių, vartojimą, gausų valgomosios druskos vartojimą, joduotos druskos... [toliau žr. visą tekstą]

Magistro darbe analizuojamas informacinių technologijų taikymas Lietuvos logopedų darbe. Apie IKT ir KMP taikymą Lietuvos logopedų darbe nėra daug žinoma, nėra parengta išsamių mokslinių straipsnių ar apžvalgų, kurie nurodytų, kokiomis informacinių technologijų programomis ar priemonėmis naudojamasi logopedų darbe, kokios iš jų vyrauja ir kurios yra efektyviausios dirbant su kalbos, kalbėjimo ir komunikacijos sutrikimų turinčiais asmenimis. Praktinis tyrimo naudingumas – atskleisti informacinių technologijų taikymo realybę logopedų darbe su kalbėjimo ir kalbos sutrikimų turinčiais asmenimis. Palyginti pasiekimus šioje srityje Lietuvoje ir užsienio šalyse. Prieduose pateikiamas internetinių svetainių sąrašas, kur galima rasti logopedinėse pratybose pritaikomų lavinimo užduočių.
Master’s thesis analyzes the informational technologies application in Lithuanian speech and language therapists’ work. The aim of this research - to assess the informational and communication technology (ICTs) and computer-based speech training system (CBST) use at the speech and language therapists work in comprehensive schools who work with persons who have language, speech and communication disorders. The empirical part of the paper deals with the use of ICTs/ CBST at speech and language therapists work, their possibility of usage, the coherence between speech therapists qualification and their age, the usage of the programs for development educable self-dependent tools, speech and language therapists interest in various sources of work-related topics and others. In the questionnaire survey (in electronic form) have participated 258 speech and language therapists who work in comprehensive schools.

碩士
高雄醫學大學
藥理學研究所
99
Nephropathy is the most common symptoms found in the end-stage of patients with diabetes. Exracellular matrix accumulation (ECM) is a hallmark in the pathogenesis of diabetic nephropathy. In which, matrix metalloproteinases (MMPs) are metal ion-depent enzymes, affecting the ECM breakdown and turnover in the glomerulous. Among them, both matrix metallopeptidase 2 (MMP-2) and matrix metallopeptidase 9 (MMP-9) expressions are prominently increased in diabetic nephropathy. Beyond their direct role in ECM turnover, MMP-2 and MMP-9 can induce the release or activation of several growth factors that are associated with tubular cell proliferation and glomerular fibrosis. Streptozotocin (STZ), an anti-cancer agent, can hurt the pancreatic islet β cells, leading to failure in secreting insulin and the resulted diabetic nephropathy. This research is to investigate hetherKMUP-1xanthin derivatives, can prevent diabetic nephropathy and associated fibrosis. Male Wistar rats were randomly divided into four groups, including control (vehicle), STZ (65 mg/kg), STZ+KMUP-1 (1mg/kg) and STZ+KMUP -1 (2.5 mg/kg). The morphology of renal tissues were evaluated by H&E stain. The expression of proteins MMP-2, MMP-9, eNOS, Bcl-2/Bax, renal tissues was exposed by Western blotting technique.Results and implication: Administration of KMUP-1 inhibited STZ-induced diabetic nephropathy and fibrosis by enhancing the expression of eNOS, suppressing MMP-2 and MMP-9, increasing ratio of Bcl-2/Bax activation. KMUP-1 is suggested to be insightful in the prevention of sclerosis diabetic nephropathy and sclerosis in patients of diabetes.

碩士
高雄醫學大學
藥理學研究所碩士班
94
G protein-mediated Ca2+ sensitization in contraction of tracheal smooth muscle（TSM）is involved in Rho kinase （ROK）and Protein Kinase C（PKC）activation pathways. The expression of ROK and PKC are activated by Sphingosine-1-phosphate （S1P）and U46619. S1P, a bioactive sphingolipid metabolite, and U46619, a thromboxane A2 analogue, both can induce TSM contractions. Our laboratory has demonstrated that KMUP-1 and KMUP-3（1-100 μM）relax the contraction of rat TSM. The results indicated that the expression of ROK and PKCα induced by S1P（2 μM）or U46619（1 μM）in cultured TSM cells were dose-dependently inhibited by KMUP-1 and KMUP-3（0.1 - 100 μM）, analyzed by western blotting, except the expression of PKCα induced by S1P（2 μM）. After further investigation for the discrepancy between KMUP-1, KMUP-3, Y27632, Chelerythrine chloride, Dexamethasone, Terbutaline, 8-bromo-cGMP and 8-bromo-cAMP（10 μM）, KMUP-1 and KMUP-3 possess similar inhibition activities on the expression of ROKα and PKCα proteins as Y27632 and Chelerythrine chloride. Relative to 10μM Dexamethasone and 10μM Terbutaline, both of 10μM KMUP-1 and 10μM KMUP-3 have batter inhibitions on the expression of ROKα and PKCα protein。 In addition, the intracellular concentration of Ca2+ induced by S1P was significantly decreased by KMUP-1. However, KMUP-3 has slight inhibition on the intracellular concentration of Ca2+。 From the results, in this study, it is proposed that KMUP-1 could inhibit the contraction of airway smooth muscle by mediating the intracellular concentration of Ca2+。Those new medicines, KMUP-1 and KMUP-3, developed by our laboratory, could possess well capabilities to inhibit the contraction induced by mechanisms of Ca2+ dependent and Ca2+ sensitization due to dose-dependently inhibitions of PKCα, ROKα and intracellular concentration of Ca2+. In conclusion, KMUP-1 and KMUP-3 may indicate the batter clinical application in the treatment of asthma.

博士
高雄醫學大學
醫學研究所
99
Pulmonary artery hypertension (PAH) is a debilitating disease, the hallmark of PAH is pulmonary arterial smooth muscle cells (PASMCs) proliferation and pulmonary pressure increasing. This study investigated the mechanisms of KMUP-1 inhibited pulmonary artery (PA) contraction and proliferation　in curing PAH. In the acute model of PAH induced by thromboxane A2 (TXA2)-mimetic U46619, KMUP-1 restored blood oxygenation and relaxed vasoconstriction by enhancing endothelial nitric oxide synthase (eNOS), soluble guanylate cyclase (sGC) and protein kinase G (PKG), and inhibited phosphodiesterase-5A (PDE-5A), RhoA/ROCK expression. The inhibition of KMUP-1 on ROCK expression was waken by ODQ、L-NAME and SQ22536. In the absence or presence of U46619 (0.5 ?嵱). Similarly, in isolated pulmonary artery (PA), KMUP-1 relaxed phenylephrine (10?n?嵱)- serotonin (5-HT)- and U46619 (0.5 ?嵱)-induced vasoconstriction, the relaxation effects of KMUP-1 are waken by ODQ、L-NAME and SQ22536. In the chronic model of PAH induced by monocrotaline (MCT), KMUP-1 prevented MCT-induced PAH over long-term administration. increased eNOS expression and reduced MYPT1 (myosin phosphatase target subunit 1) phosphorylation, RhoA/ROCK, 5-HT transporter (5-HTT) expression in lung tissue, reduced plasma 5-HT increasing, PA wall thickening, proliferation and right ventricular hypertrophy (RVH). Incubating PASMCs with KMUP-1 inhibited thapsigargin-induced Ca2+ efflux and angiotensin II-, 5-HT-induced Ca2+ influx. KMUP-1 (1-100 ?嵱) inhibited 5-HT-induced RhoA/ROCK expression, while KMUP-1, Y27632 and simvastatin at 10 ?嵱 inhibited 5-HT-induced 5-HTT expression and KMUP-1 inhibited 5-HT-induced phosphorylation of AKT and ERK1/2 in PASMCs. KMUP-1 (1-100 ?嵱) concentration-dependently increased the expression of eNOS and 5-HT2B and production of NO in human pulmonary arterial endothelial cells (HPAEC), 5-HT2B receptor antagonist SB200646 decreased the effects of KMUP-1 on eNOS. In radioligand binding, binding ability of KMUP-1 was 5-HT2B > 5-HT2A > 5-HT2C. KMUP-1 inhibits MCT-induced PA proliferation by binding to 5-HT2A, 5-HT 2B and 5-HT 2C receptors, increasing endothelial eNOS/5-HT2B receptor expression and inhibiting 5-HTT/RhoA/ROCK expression and AKT/ERK phosphorylation. These findings suggest that KMUP-1 relieves and prevents PAH via enhancement of eNOS to releases NO and create a cGMP-dependent inhibition of RhoA/ROCK and Ca2+ desensitization in PASMCs. KMUP-1 inhibits MCT-induced PA proliferation by binding to 5-HT2A, 5-HT2B and 5-HT2C receptors, increasing endothelial eNOS/5-HT2B receptor expression and inhibiting 5-HTT/RhoA/ROCK expression and AKT/ERK phosphorylation. In conclusion, KMUP-1 has the potential to reduce vascular resistance, remodeling, hyperplasia and RVH in treating PAH.

碩士
高雄醫學大學
藥理學研究所
100
Bone mass of human is controlled by continuous bone remodeling through osteoblastic bone formation and osteoclastic bone resorption. Abnormality in bone remodeling resulting in a variety of bone disorders such as osteoporosis. In the field of bone-decreasing diseases, there has been growing interest in anabolic agents that enhance bone formation. Recent studies suggest that phosphodiesterase (PDE) 4 inhibitors have therapeutic effects in different experimental osteopenia models. Our previous studies show that KMUP-1, a novel nitric oxide enhancer developed by our laboratory, has inhibitory effects on the phosphodiesterase (PDE). Thus, this study aimed to investigate the effect of KMUP-1 on differentiation of osteoblasts, and to elucidate the cellular and molecular mechanisms involved. Primary osteoblasts and osteoblastic MC3T3-E1 cells were examined in this study. In vitro, results showed that KMUP-1 up-regulated alkaline phosphatase (ALP) activity, which is the main marker of osteoblastic differentiation and increased the formation of mineralization without any evidence of cytotoxicity. Moreover, KMUP-1 enhanced the mRNA expression of the osteoblastic differentiation markers, collagen type Ιa (ColIa1), ALP osteocalcin (OCN), and Runx2, which is the key transcription regulator for osteoblastic differentiation. We also found that KMUP-1 activated the BMP/Smad and Wnt/β-catenin signaling pathways through inhibited PDE, stimulated PKA and PKG activation. These findings collectively suggested that KMUP-1 stimulates osteogenic differentiation and enhances mineralization in osteoblasts probably through activation of PKA and PKG, then stimulated the BMP/Smad and Wnt/β-catenin signaling pathways in vitro. Thus, KMUP-1 may have a role in the prevention and treatment of bone-decreasing diseases.

碩士
高雄醫學大學
藥理學研究所
101
Autophagy is an evolutionary conserved process involved in the degradation of long-lived proteins and excess or dysfunctional organelles in eukaryotic cells. In the heart, autophagy functions predominantly as a pro-survival pathway during cellular stress by removing protein aggregates and damaged organelles, protecting the heart against famine, ischemia/reperfusion and heart failure. Cardiac remodeling involves increased rates of cardiomyocyte cell death and precedes heart failure. Meanwhile, cardiac fibroblasts play important role by regulating structure, biochemical, mechanical and electrical propertied of the heart. Therefore, we aimed to investigate signal transduction pathways involved in the induction of myocardial autophagy by KMUP-3. KMUP-3 is a chemically synthetic xanthine-based derivative. It has been demonstrated to have phosphodiesterase inhibition, endothelial nitric oxide synthase (eNOS) enhancement and KATP channel opening activities. When cardiac fibroblasts and cardiomyocytes were treated with KMUP-3 for different time, we found that the expression of LC3-II protein and autophagy-related genes including Beclin-1 and Atg7 were markedly upregulated in time- and dose-dependent manner. In addition, cells treated by KMUP-3 developed autophagosome-like characteristics, including single and double membrane vacuoles containing intact and degraded cellular debris. To investigate the ability of KMUP-3 to stimulate autophagy as a potential mechanism involved in eNOS phosphorylation. We treated cells with NOS inhibitor L-NAME, and found that L-NAME significantly inhibited KMUP-3-induced LC3-II expression and eNOS phosphorylation. Because it was recently reported that phosphatidylinositol 3-kinase (PI3K) and its downstream effector Akt are implicated in the activation of eNOS. We treated cells with PI3K inhibitor wortmannin, and found that wortmannin abolished KMUP-3-induced eNOS phosphorylation and Akt phosphorylation, providing the first evidence that KMUP-3 stimulates autophagy probably through PI3K/Akt/eNOS signaling. Besides, enhancement of AMPK activity also induces eNOS activity. We treated cells with KMUP-3 for different time, and found that the phosporylation of AMPK was markedly upregulated in time-dependent manner. Taken together, our results show that KMUP-3 induces autophagy through AMPK/PI3K/Akt/eNOS signaling in cardiac fibroblasts and cardiomyocytes.

碩士
高雄醫學大學
藥理學研究所碩士班
94
KMUP-1, a chemically synthetic xanthine-based derivative, has been demonstrated not only increase of cyclic nucleotides and inhibition of phosphodiesterases, but also activation of K+ channels resulting in relaxation in rat aortic smooth muscle (SM), rabbit corpus cavernosum and guinea-pig tracheal SM. However, a direct evidence of calcium currents inhibition by KMUP-1 has not yet been documented. This study is to examine the effects of KMUP-1 on L-type calcium currents (ICa,L). We used the conventional whole cell patch-clamp technique to investigate Ba2+ currents (IBa) through L-type Ca2+ channels in rat basilar artery myocytes. Under voltage-clamp conditions, KMUP-1 inhibited the IBa in a concentration-dependent manner without any change in current-voltage relationship of IBa. Additionally, KMUP-1 inhibited the protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA, 1 ?嵱), induced IBa. Pretreatment with the PKC inhibitor chelerythrine (5 ?嵱) enhances the inhibition of IBa by KMUP-1. However, a Rho kinase inhibitor Y-27632 (30 ?嵱) failed to affect the inhibition of IBa by KMUP-1. In fura-2-loaded rat basilar arteries, KMUP-1 inhibited the Ca2+ signal evoked by 100 mM KCl and 1 ?嵱 PMA. In addition, KMUP-1 also inhibited the Ca2+ signal evoked by 10 ?嵱 thapsigargine, a specific inhibitor of endoplasmic reticulum Ca2+-ATPase. In light of these results, we suggest that KMUP-1 inhibits the L-type calcium channels in concentration- and voltage-dependent manners in rat basilar artery myocytes and the effects may partially related to the PKC pathway.

碩士
高雄醫學大學
醫學研究所碩士班
93
Phosphodiesterase inhibitor had been proved on its anti-angiogenesis and anti-proliferation effects through numerous signal transduction pathway in various cancer cells. In this study, KMUP-1, a phosphodiesterase inhibitor, was investigated on its anti-angiogenesis and anti-proliferation effects in human liver cancer cell line Hep G2 . The cell growth inhibition ability of KMUP-1 was determined by MTT assay. The results indicated that KMUP-1, possessed a higher cell growth inhibition ability in normoxic condition than in hypoxic condition. Using flow cytometry techniques, we found that KMUP-1 induced apoptosis in a time- and concentration dependent manners under normoxic and hypoxic state, and these effects was dependent on sGC/cyclic GMP/PKG signal pathway. Notably, the effects were dramatically changed after 48 and 72 hrs exposured to KMUP-1. In addition, we also found that KMUP-1 could arrest cell cycle progression at G0/G1 phase. Western blotting analysis indicated that KMUP-1-induced cell cycle arrest may depend on increasing p21CIP1/WAF1 and p27KIP1 protein expression and on decreasing cyclin D/CDK6 or cyclin D/CDK4 complex expression. Hypoxia-inducible factor-1 alpha (HIF-1α), a component of HIF-1, is expressed in survived human tumors and renders cells grown under hypoxia condition. Besides, HIF-1 is a key transcription factor that regulate the blood supply through activity on the expression of vascular endothelial growth factor (VEGF), which promotes the angiogenesis. cause significant decrease on HIF-1α and VEGF protein expression. In summary, KMUP-1 could not only activate cyclin kinase inhibitors (CKIs) expression and inhibit cyclin D/CDK4 or 6 complex level for arresting hepatoma cells growth but also cause cell apoptosis dependent on sGC/cGMP/PKG signal pathway both under normoxic and hypoxic condition. In addition, KMUP-1 can reduce HIF-1α and VEGF expression under hypoxic condition. These findings implicate that KMUP-1 may be useful for anti-angiogenesis and anti-proliferation therapy in human liver cancer.The result from Western blotting analysis also showed that KMUP-1 could

碩士
高雄醫學大學
藥理學研究所
98
Hepatic ischemia-reperfusion (I/R) -induced injury is a major cause of morbidity and mortality associated with liver transplantation and resectional surgery as well as septic and hemorrhagic shock. Indeed, there is a large body of experimental and clinical evidence suggesting that I/R induced by these surgical procedures or pathophysiological events injures the liver and may ultimately lead to tissue dysfunction and possibly liver failure. These surgical procedures or pathophysiological events usually results in oxidative stress, hepatocyte death, endothelial cell damage and microcirculatory disturbances, leading to liver dysfunction. Previous reports have defined this injury as bi-phasic, having both an acute and a sub-acute phase. The acute phase occurs within the first 6 h of reperfusion and is characterized by the activation of resident Kupffer cells, resulting in enhanced production of reactive oxygen species (ROS). Historically, enhanced ROS production has been thought to generate severe oxidative stress to the tissue by virtue of its ability to degrade membrane lipids and/or proteins. A growing body of experimental evidence suggests that nitric oxide (NO) may also modulate I/R-induced tissue injury in various organ systems. In vitro and in vivo data suggest that endothelial nitric oxide synthase (eNOS)-derived NO may act to protect tissue by virtue of its ability to react with and decompose ROS, and interfere with Caspase activation. Guanosine 3’,5’cyclic momnophosphate (cGMP), a key intracellular second messenger molecule, has been shown to up-regulate Bcl-2 expression in some cells. Phosphodiesterase 5 (PDE5) inhibition results in raising the intracellular cGMP concentration which subsequently activates cytosolic cGMP-dependent protein kinase (PKG). Previous studies demonstrated that PKG expression increased the protective effect against necrosis and apoptosis following simulated ischemia and reoxygenation. KMUP-1, a xanthine derivative, was demonstrated to promote vasodilation, inhibition of PDEs, enhancement of cGMP, and activation of K+ channels. In this study, we investigated the protective effects and pharmacological mechanism of action of KMUP-1 in liver after I/R. Male Wistar rats were randomly divided into five groups, as follows: Sham-operated group, I/R with Vehicle group, I/R with KMUP-1 0.25 mg/kg group, I/R with KMUP-1 0.5 mg/kg group, and I/R with KMUP-1 1 mg/kg group. They underwent 70% partial hepatic ischemia for 45 minutes and subsequent reperfusion for 240 miniutes. Sham-operated group underwent all surgical procedure except ligation of portal triad. All rats were treated by bolus injection via femoral vein (i.v.) 10 min before partial ischemia. They were sacrificed at 240 minutes after reperfusion. In a rat model of acute hepatic ischemia-reperfusion, administration of KMUP-1 inhibited I/R-induced apoptosis, as detected by ladder-pattern fragmentation of genomic DNA. KMUP-1 improved parameters of liver function, enhanced protein expressions of eNOS/ cGMP/ PKG pathway, and ratio of Bcl-2/ Bax and decreased ROS production and caspase-3 activation. In conclusion, these results suggest that KMUP-1 may protect liver from I/R-induced apoptosis by scavenging free radicals and regulating the protein expression of Bcl-2 family and eNOS/ cGMP/ PKG pathway. Thus, KMUP-1 will be useful clinically in the prevention of acute hepatic ischemia-reperfusion injury.

碩士
高雄醫學大學
藥理學研究所碩士班
95
In pulmonary vascular smooth muscles, modulation of large-conductance Ca2+-activated K+ (BKCa) channel is important in the regulation of pulmonary arterial pressure. KMUP-1, a synthetic xanthine-based derivative, has been demonstrated to have BKCa currents activation in basilar artery myocytes. In this study, we tried to investigate the ionic mechanisms of KMUP-1 whether and by what signaling to stimulate BKCa in pulmonary arteries. Pulmonary arterial smooth muscle cells (PASMCs) were enzymatically isolated from Sprague-Dawley rats. KMUP-1 significantly increased the BKCa current and open probability using conventional whole cell and inside-out patch-clamp techniques. Increased BKCa current activity was abolished by a BKCa channel inhibitor iberiotoxin (100 nM). To study the mechanisms of KMUP-1 on BKCa channels, a soluble guanylate cyclase inhibitor (ODQ, 10 μM), an adenylate cyclase inhibitor (SQ22536, 10 μM), competitive antagonists of cGMP and cAMP (Rp-cGMP, 100 μM and Rp-cAMP, 100 μM), and cGMP- and cAMP-dependent protein kinase inhibitors (KT5823, 300 nM and KT5720, 300 nM) were applied. BKCa current activation by KMUP-1 was significantly inhibited by these agents. We also found that KMUP-1 has the ability to prevent uridine triphosphate (UTP, 10 μM) -induced inhibition of delayed rectifier K+ (KDR) channels. To study the relationship of KMUP-1 and RhoA/Rho kinase pathway on PASMCs, a RhoA inhibitor C3 exoenzyme (10 μg/ml), a Rho kinase (ROCK) inhibitor Y27632 (30 μM), and PKG and PKA inhibitors (KT5823, 300 nM and KT5720, 300 nM) were applied. In conclusion, KMUP-1 increases the BKCa current and channel open probability by stimulating the activity of cyclic nucleotide-dependent protein kinases. Additionally, KMUP-1 also inhibits the RhoA/ROCK pathway would result in the relaxation of pulmonary arteries.

碩士
高雄醫學大學
藥理學研究所碩士班
94
Cerebral vasospasm after aneurismal subarachnoid hemorrhage (SAH) is characterized by diffuse and long-lasting narrowing of arteries. The large conductance Ca2+-activated K+ (BKCa) channel plays a negative feedback role to prevent vasospasm. KMUP-1, a xanthine-based derivative, has been demonstrated to activate K+ channels resulting in the relaxation of aortic and tracheal smooth muscles. In this study, we tried to investigate whether SAH-induced vasospasm could be a defect in the function of vascular BKCa channels, and therefore to explore the protective activity of KMUP-1 in SAH rats. Rats were divided into four groups including control, SAH, KMUP-1-treated and pinacidil-treated groups. Rats were subjected to experimental SAH by injecting autologous blood into the cisterna magna, then KMUP-1 (1 mg/kg) and pinacidil (1 mg/kg) were administered intraperitoneally at 1 h after SAH in this study. All rats were sacrificed at 24 h after SAH in this study. Cerebrovascular smooth muscle cells (CVSMCs) were enzymatically isolated from rat basilar arteries. Using whole-cell patch-clamp techniques, the iberiotoxin (IbTX)-sensitive BKCa currents attenuated in SAH groups compared with control groups. Pretreatment with KMUP-1 or pinacidil, the IbTX-sensitive currents were restored. In inside-out patches, the unitary conductance and voltage sensitivity of SAH, KMUP-1 and pinacidil groups showed effects similar to the results of control groups. However, the BKCa channel open probability (NPo) and calcium sensitivity were decreased in SAH groups compared with control groups. In conclusion, we suggest that the down-regulation of the BKCa channel activity could be due to the modification of the BKCa channel’s calcium sensitivity. From the results indicated that KMUP-1 and pinacidil could protect against SAH-induced vasospasm.

碩士
高雄醫學大學
藥理學研究所碩士班
94
It is known that chronic treatment of a ??-adrenergic agonist, isoproterenol, induced cardiac hypertrophy. Another model, spontaneously hypertensive rats (SHR) also developed chronic hypertension leading to cardiac hypertrophy and heart failure. When given of NOS inhibitor N-omega-nitro-L-arginine (L-NNA) or KATP channel blockor 5-hydroxydecanoate (5-HD), aggravated the response of cardiac hypertrophy. The present study investigated that the in vivo effects of KMUP-1 and sildenafil, phosphodiestrase-5 (PDE-5) inhibitors, on the hypertrophic responses of rat heart to isoproterenol and SHR, and the relation of the effects to the levels of myocardial cyclic guanosine monophosphate (cGMP) and nitric oxide, the activities of PKG, NOS, GSK3-???z?ncalcineurin A and ERK1/2. The results showed that daily subcutaneous administration of isoproterenol for 10 days caused significantly cardiac hypertrophy, cell injury and decline in survival. Treatment of L-NNA (20 mg/l/day) in SHR also developed significantly cardiac hypertrophy and decline in survival. KMUP-1 (0.5 mg/kg/day) and sildenafil (0.7 mg/kg/day) were intraperitoneal injected, one hour before isoproterenol. KMUP-1 and sildenafil was also intraperitoneal injected to SHR. We found that both KMUP-1 and sildenafil significantly improved the survival rate and decreased the ratio of heart weight to body weight (HW/BW). Both KMUP-1 and sildenafil increased the expression of eNOS, PKG and GSK-3??, and the production of NO and cGMP, but suppressed the expression of iNOS, calcineurin A and ERK1/2. However these effects of KMUP-1 and sildenafil were partially reversed by treatment of L-NNA and 5-HD. These present study indicated that KMUP-1 similar to sildenafil has a cardioprotective effect against isoproterenol- and spontaneously hypertension-induced myocardial cell injury. KMUP-1 may through the activation NO/cGMP/PKG-1 pathway and then to improve survival rate and cardiac function, suggesting that it may have great potential in the prevention of cardiac hypertrophy and heart failure.

碩士
高雄醫學大學
藥理學研究所碩士班
95
Cardiac myocytes undergo apoptosis under condition of ischemia. Myocardial oxidative stress and Ca2+ overload induced by ischemia may be involved in the development and progression of myocardial apoptosis. We hypothesized that KMUP-1 could prevent the ischemia-induced reactive oxygen species production and Ca2+ overload, leading to decreased calcium-responsive signaling in myocardial dysfunction. We showed that serum/glucose deprivation and hypoxia, components of ischemia in vivo, resulted in apoptosis of H9c2 cardiomyoblasts. H9c2 cells apoptosis is evidenced by an increase in DNA ladder and Hoechst 33342 positive. In this model of simulated ischemia, represented by serum/glucose deprivation and hypoxia, KMUP-1 (0.1, 1, and 10 μM) could protect H9c2 cells against simulated ischemia-induced apoptosis. KMUP-1 also inhibited several of the molecular events of apoptosis that follow simulated ischemia, such as the DNA fragmentation, the production of ROS, and the ratio of Bcl-2 to Bax. Furthermore, KMUP-1 could modulate eNOS and enhances NO production. KMUP-1 also could stimulate soluble guanylate cyclase (sGC) and activate PKG. NO has been shown to exert a number of actions that would be expected to be beneficial during myocardial ischemia, including inhibition of Ca2+ inflow and a decrease in myocardiac calculated oxygen consumption. Moreover, our results suggested that KMUP-1 can not only increase the expression of ERK, but can also decrease the expression of JNK and p38. In addition, pretreatment with KMUP-1 could protect against LPC-induced cytotoxicity in H9c2 cells and the drug can also inhibit LPC-stimulated ROS production, intracellular calcium elevation, the decrease of ERK phosphorylation, and the increase of p38 phosphorylation. In conclusion, these results demonstrate that KMUP-1 can increase the survival rate of H9c2 cells in the hypoxia and LPC models. The cardioprotective effect of KMUP-1 may via MAPK and NO-cGMP-PKG pathway, decrease intracellular ROS production, and Ca2+ overload to prevent myocardial ischemic injury.

碩士
高雄醫學大學
藥理學研究所
97
Exposure to hypoxia results in the development of pulmonary hypertension An increase in the intracellular Ca2+ concentration ([Ca2+]i) in pulmonary artery smooth muscle cells (PASMCs) is a major trigger for pulmonary vasoconstriction and proliferation. The aim of this study was to examine the mechanism by which KMUP-1 inhibited the hypoxia-induced canonical transient receptor potential (TRPC) protein overexpression and regulated the [Ca2+]i through store-operated calcium channels (SOCC). Primary PASMCs were cultured from Spraque-Dawley rats and placed in a modular incubator chamber under 1% O2/5% CO2 for 24 hours to induce SOCC overexpression. KMUP-1 (1 μM) inhibited hypoxia-induced TRPC family encoded for SOCC overexpression, particularly TRPC1. KMUP-1-inhibited TRPC1 attenuated by the PKG inhibitor KT5823 (1 μM) and the PKA inhibitor KT5720 (1 μM). A PKC activator PMA (1 μM)-mediated TRPC1 was attenuated by KMUP-1. Taken together, we suggest that the effects of KMUP-1 on TRPC1 might involve the activation of cGMP/PKG and cAMP/PKA pathways, and inhibition of PKC pathway. Moreover, we use Fura-2/AM to measure the store calcium release from sarcoplasmic reticulum (SR) and calcium entry through SOCC in hypoxic PASMCs. Under hypoxic conditions, the activity of store-operated calcium entry (SOCE) was enhanced. KMUP-1 could refill the stores and attenuate SOCE through SOCC in hypoxic PASMCs. In conclusion, KMUP-1 inhibited TRPC1 protein expression and reduced CCE could be through the SOCC in hypoxic PASMCs.

碩士
高雄醫學大學
藥理學研究所
97
Endothelin-1 (ET-1) has been implicated in cardiac pathology, such as the progression from cardiac hypertrophy to failure. KMUP-1 is a unique xanthine and piperazine derivative, combining the sGC stimulation, K+ channels opening and particularly phosphodiesterase type 5（PDE-5）inhibition activities in one molecule. The purpose of this study was to determine the efficacy and the possible mechanism of action of KMUP-1 on the ET-1-induced cardiac hypertrophy in cardiomyocytes. When cardiac myoblasts (H9c2 cells) were treated with ET-1 (100 nM) for 4 days, hypertrophy was observed, as assessed by the measurement of cell surface area, and this effect was strongly prevented by KMUP-1. Western blot analysis showed that KMUP-1 decreased ET-1-induced phosphorylation of ERK1/2, p38 and Akt/GSK3?? and also decreased calcineurin/NFAT and RhoA/Rock expression. KMUP-1 also enhanced HO-1 protein expression, and previous study have shown that HO-1 can inhibit mitogen-activated protein kinase (MAPK), calcineurin/NFAT signaling and hypertrophy in cardiac myocytes. Electrophoretic mobility shift assay showed that KMUP-1 inhibited ET-1-induced activator protein-1 (AP-1) DNA binding activities. In addition, incubation of H9c2 cells with KMUP-1 significantly decreased the intracellular peroxide level induced by ET-1 according to fluorescent microscopic observation using DCHF-DA as a fluorescent substrate. We further examined the effect of KMUP-1 on ET-1-induced increase oxygen consumption as an index of ROS generation in H9c2 cells. In conclusion, these findings suggest that KMUP-1 protects ET-1-induced hypertrophy in H9c2 cells by blocking phosphorylation of ERK1/2, p38, Akt/GSK3β, calcineurin/NFAT and RhoA/Rock activation, enhancing HO-1 protein expression and decreasing ROS generation.

碩士
高雄醫學大學
藥理學研究所
96
Pulmonary artery hypertension (PAH) is characterized by a sustained increase in pulmonary artery pressure (PAP) leading to progressive right ventricular failure and death. This study is to investigate the effects of KMUP-1 on PAP and potassium channels in monocrotaline (MCT) -induced PAH rats. Sprague-Dawley (SD) rats were divided into three groups including the control (CTL), MCT, and KMUP-1 treated. PAH rats were induced by single intraperitoneal injection of MCT (60 mg kg-1), then KMUP-1 (5 mg kg-1) was administrated intraperitoneally once daily from 2nd day to 20th day. All rats were sacrificed at 21th day after treating MCT. The right ventricular systolic pressure (RVSP) and arterial pressure were determined simultaneously from right ventricle and femoral artery catheterization, respectively. Pulmonary artery smooth muscle cells (PASMCs) were enzymatically isolated from intrapulmonary arteries. The currents of large conductance calcium activated potassium (BKCa) channel and voltage-gated potassium (Kv) channel were measured by using conventional whole cell or inside-out patch-clamp techniques. In this study, the increase of RVSP was almost fully prevented by KMUP-1. Using whole cell patch clamp, the paxilline-sensitive BKCa and 4-aminopyridine (4-AP) sensitive Kv currents were attenuated in MCT group compared with the CTL group. Pretreatment with KMUP-1 (5 mg kg-1) reversed the inhibition of paxilline-sensitive and 4-AP sensitive currents. In inside-out patch clamp, the unitary conductance and voltage sensitivity of MCT and KMUP-1 treated groups are similar to CTL group. However, the open probability (NPo) and calcium sensitivity of BKCa channels were decreased in MCT group compared with CTL group. Pretreatment with KMUP-1 also reversed the MCT-induced reduction of NPo and calcium sensitivity. In conclusion, persistent decrease of the BKCa and Kv currents may take part in the development of MCT-induced PAH rats. KMUP-1 did decrease the elevated RVSP and prevent downregulation of BKCa and Kv channels in MCT rats. The results provide the evidence that KMUP-1 could be used in the management of PAH in the future.

碩士
高雄醫學大學
醫學系藥理學科研究所
102
Autophagy is important for the turnover of organelles at low basal levels under normal conditions and it is upregulated in response to stresses such as ischemia/reperfusion and in cardiovascular diseases such as heart failure. Apoptosis also plays important biological roles in the pathogenesis of many diseases. Oxidative stress induced by myocardial infarction is one of the major factor of heart failures. In our previous studies, 7-[2-[4-(4-nitrobenzene)piperazinyl]ethyl]-1,3-dimethylxanthine (KMUP-3) is a chemically synthetic xanthine-based derivative. It has been shown to induce autophagy in cardiac fibroblasts. The aim of this study was to investigate how KMUP-3 modulates hydrogen peroxide (H2O2)-induced apoptosis in neonatal rat cardiac fibroblasts and to elucidate the cellular and molecular mechanism. 　　In this study, MTT assay showed that H2O2 induced cardiac fibroblasts death, and pre-treat KMUP-3 block it. Flow cytometry analysis H2O2 induced cardiac fibroblasts death through apoptosis, and pre-treat KMUP-3 prevent cell death. Western blot showed that KMUP-3 enhanced p-eNOS, eNOS, PKG, LC3-II, Atg7 formation, and increased Bcl-2/Bax ratio in H2O2-treated neonatal rat cardiac fibroblasts. Moreover, KMUP-3 attenuated H2O2-induced MMP-2, MMP-9 and cleaved caspase-3 protein expressions. These effects were blocked by both the L-NAME and L-NIO, indicating that eNOS plays a role in the modulation of KMUP-3 in H2O2-induced apoptosis. However, when cardiac fibroblasts treated with Atg7 siRNA, which blocked the autophagy in the cells and resulted in a further increase in cell apoptosis. 　　These results showed that KMUP-3 may promote autophagy to decrease oxidative stress-induced apoptosis in cardiac fibroblasts. KMUP-3 might exert cardioprotective effects in heart diseases through regulation of autophagy and apoptosis. These cardioprotective effects are possibly mediated through e-NOS enhancing and cGMP/ PKG-dependent signalling pathways.

Rhythmic motions, regular or irregular, are an integral part of motor behavior both in health and in disease. Better understanding of its neural control mechanisms helps in developing methods for controlling the progression of diseases manifesting as rhythmic motions. I studied two specific aspects of rhythmic motions: bilateral coordination of hand tremors in human subjects and modular control of locomotion in invertebrates. Many types of tremors, including the physiological tremor (PT) and the essential tremor (ET) occur in limbs on both the sides of the body, with similar fundamental frequency of the oscillation. This raises the possibility that the contralateral tremors may have a common source or are otherwise coupled. However, while significant contralateral interaction is seen in these two types of tremors, only limited evidence of bilateral coherence has been shown in the previous literature. Therefore, in my study I explored the existence of a weak coupling between the left and right oscillators the may lead to intermittent bilateral coherence. I measured triaxial acceleration of the two hands and systematically assessed their bilateral coherence, using both stationary and non-stationary (wavelet-based) analyses methods. Measuring all three axes allowed examination of a more complete set...

碩士
高雄醫學大學
藥理學研究所
98
Serotonin (5-hydroxytryptamine, 5-HT) is a potent pulmonary vasoconstrictor and promotes pulmonary arterial smooth muscle cells (PASMCs) proliferation. K+ channels play an essential role in regulating resting membrane potential and contraction of vascular smooth muscle. Serotonin-induced inhibition of K+ channels could increase the intracellular Ca2+ concentration ([Ca2+]i) in PASMCs and could be a major trigger for pulmonary vasoconstriction and development of pulmonary arterial hypertension (PAH). In this study, we examined the mechanism of action by which KMUP-1 could attenuate the pulmonary vasoconstriction in isolated pulmonary arteries (PAs) and prevent the serotonin-induced K+ channel inhibitory activity in PASMCs. PASMCs were primary cultured from Spraque-Dawley rats and placed in the incubator. PASMCs from passage 3 to 6 were used in this study. Cells were incubated with serotonin (10 μM), KMUP-1 (1 μM) or test agents in the same medium for 24 h. Stimulation of isolated PAs with serotonin induced a significant contractile response and KMUP-1 could prevent it. The various K+ channel inhibitors blocked the effect of vasodilatation of KMUP-1. We suggest that the prevention of KMUP-1 on serotonin-induced contraction might via activation of K+ channels. Additionally, serotonin caused decreases the protein expression and activity of voltage-gated K+ (Kv1.5 and Kv2.1) and large-conductance Ca2+-activated K+ (BKCa) channels in PASMCs. KMUP-1 avoided serotonin-induced decreases in K+ channel proteins. Serotonin-inhibited K+ channel proteins were attenuated by the PKA activator 8-Br-cAMP (100 μM). And a PKC inhibitor chelerythrine (1 μM) enhanced serotonin-inhibited K+ channel proteins. These results indicate that serotonin-inhibited K+ channel might involve the PKA inhibition and PKC activation. In conclusion, KMUP-1 could be used to prevent serotonin-induced K+ channels inhibition and vasoconstriction, which promote the development of PAH.

碩士
國立成功大學
資訊工程學系碩博士班
96
The usage of the Internet has grown enormously in recent years. There are many viruses affect the network security significantly. Network Intrusion Detection System (NIDS) is a system developed for identifying attacks by using a set of rules. These rules are defined by Snort; one of the most often applied NIDS in the world. Searching for multiple patterns is a computationally expensive task in NIDS. We want to find a method to search patterns more efficiently and have cheaper cost. However, the traditional software-based NIDS solutions usually can not achieve a high-speed required for ever growing Internet attacks. Therefore, the issue of the implement in hardware for preventing the malicious attack from intruders has became more significantly. In this thesis, we proposed an efficient architecture based on KMP algorithm. Our main idea is design an architecture that always processes n characters per clock cycle. Here the variable, n, means the amount of the input characters we could process in each clock cycle. To ensure our proposed scheme could always process efficiently, we discuss all possible circumstances and compare at most two potential matching prefix at the same clock cycle. Although there will be many complex compare results, our design will find the correct index of the pattern and repeat the comparisons in next clock cycle. According to our observation, our approach could process in giga-bit ratio and use a small quantity of hardware resource.

碩士
高雄醫學大學
藥理學研究所
99
Pancreatic β-cells release insulin depending on glucose metabolism and membrane depolarization. Insulin secretion is triggered by the closure of ATP-sensitive K+ (KATP) channels, causing a membrane depolarization and Ca2+ influx. Insulin release process is terminated by membrane repolarization via opening of voltage-dependent K+ (Kv) channels. KMUP-1, a chemically synthetic xanthine-based derivative, has been demonstrated not only effective on K+ channels, but also a phosphodiesterases inhibitor. However, it has newer been addressed in the inhibition of Kv channels in pancreatic β-cells. In this study, we examined the action mechanisms by which KMUP-1 could inhibit high glucose (25 mM)-induced Kv current activation in pancreatic β-cells. Pancreatic islets were isolated from Wistar rats. For electrophysiological study, islets were dispersed into single β-cells by 0.05% trypsin-EDTA solution and plated onto 35-mm culture dishes for 2-4 days. Perforated patch-clamp (nystatin 200 μg/mL in pipette solution) technique was used to investigate Kv currents. Pancreatic β-cells were identified by the size, capacitance and membrane potential. The peak Kv current in 25 mM glucose-treated β-cells was ~1.4-fold greater than in 5.6 mM glucose (normal)-treated, measured at +50 mV. KMUP-1 (1, 10, 30 μM) prevented 25 mM glucose-stimulated Kv currents in a concentration-dependent manner. Insulin secretion was decreased after high glucose incubation, and increased by treating KMUP-1. Reduction of high glucose-mediated Kv current was found in incubating protein kinase A (PKA) activator 8-Br-cAMP (100 μM). Additionally, KMUP-1 (30 μM)-inhibited this current was partially reversed by the PKA inhibitor H-89 (1 μM). Otherwise, pretreatment with PKC activator or inhibitor had no effect on Kv currents in normal or high glucose condition. In conclusion, glucose-stimulated insulin secretion was reduced by the Kv channels opening. KMUP-1 could decrease high glucose-stimulated Kv currents via the PKA but not PKC signaling pathway. According to these results, we provide an evidence that KMUP-1 might be useful in the control of type II diabetes.

碩士
高雄醫學大學
藥理學研究所
100
Neuropathic pain is a deleterious change to the sensory transmission pathway after a lesion or dysfunction of the nervous system. Peripheral neuropathic pain is characterized by spontaneous pain, hyperalgesia and allodynia. It adversely affects quality of life, reduces physical and emotional functioning. Previous studies have been shown that peripheral nerve injury could induce inflammatory states, resulting in the genesis and maintenance of neuropathic pain. Phrmacological interventions have been tried in experimental models and in clinical trials but unfortunately have met with limited success. In the present study, we tried to investigate whether KMUP-1 could improve pain hypersensitivity and reduce the production of inflammatory mediators, and also explore the possible underlying mechanism in sciatic nerve in rats with chronic constriction injury (CCI) to induce neuropathic pain. Sprague–Dawley (SD) rats were randomly divided into four groups, including sham, sham with KMUP-1, chronic constriction injury (CCI) of bilateral sciatic nerve and CCI with KMUP-1 group. Rats were subjected to CCI surgery, KMUP-1 (5 mg/kg) was administrated intraperitoneally once daily starting at 1 day after surgery. Both mechanical and thermal responses of both hind paws were assessed before surgery and at day 3, 7, 14 after sciatic nerve injury. Behavioral monitoring, including thermal hyperalgesia and mechanical allodynia were both significantly reduced in CCI treated with KMUP-1 group compared with CCI group. Sciatic nerves were isolated from the four groups for western blots and enzyme-linked immunosorbent assay (ELISA) to analyze proteins and cytokines level, respectively. We found that the protein expressions of inflammatory mediators (COX2, iNOS, nNOS) and proinflammatory mediators (IL-1β, TNF-α) induced by CCI were significantly decreased in KMUP-1-treated group at day 7 after surgery. Additionally, KMUP-1 inhibited neuropathic pain-related mechanisms, including p38 and ERK activation, but not JNK, which are members of mitogen-activated protein kinases (MAPKs). KMUP-1 also blocked CCI-induced inhibitor κBα (IκBα) phosphorylation and nuclear factor-kappa B (NF-κB) translocation to nuclear. In conclusion, the results suggest that KMUP-1 has anti- inflammatory and anti-pain hypersensitivity properties in CCI-induced neuropathic pain through inhibition of MAPK and NF-κB activation. Therefore, KMUP-1 might be a potential agent for the control of neuropathic pain.

碩士
高雄醫學大學
藥理學研究所
101
Chronic neuropathic pain is a refractory pain characterized by its complex mechanisms and diverse clinical manifestations. Traditional therapies usually bring about many side effects and limited success. In the study, we attempt to investigate whether KMUP-1 could reduce hyperalgesia and inflammatory mediators, and to reveal its underlying mechanisms in the dorsal root ganglion (DRG) following chronic constriction injury (CCI)-induced neuropathic pain. Sprague–Dawley rats were randomly divided into four groups: sham, sham with KMUP-1 (5 mg/kg, i.p), CCI and CCI with KMUP-1 (5 mg/kg, i.p). KMUP-1 (5 mg/kg) was administrated intraperitoneally once daily starting at day 1 after CCI surgery. Each group of rats (n=5) were sacrificed and L4-L6 DRGs removed quickly at day 3, 7 and 14 after CCI. Behavior tests were assessed before surgery and at those scheduled time-points after CCI. KMUP-1 decreased mechanical allodynia at day 3, 7 and 14, and thermal hyperalgesia at day 7 and 14 after CCI ipsilateral side, but not CCI contralateral side. KMUP-1-treated group significantly inhibited CCI-induced inflammatory mediators (iNOS, COX2) and proinflammatory mediators (TNF-??, IL-1??). Activation of PKA, PKC and ERK in the DRG contributes to the initiation of CCI-induced pain hypersensitivity. KMUP-1 inhibited the BDNF, CGRP, PKA, PKC and ERK activations that could attribute, at least in part, to its possible mechanisms in CCI-induced neuropathic pain. KMUP-1 also blocked CCI-induced inhibitor ?羠?? (I?羠??) phosphorylation and nuclear factor-kappa B (NF-?羠) translocation to nuclear. Based on our results, KMUP-1 has anti-inflammatory and anti-hyperalgesia properties in CCI-induced neuropathic pain via inhibition of BDNF, CGRP, PKA, PKC and NF-?羠. We suggest that KMUP-1 might be a potential agent for the control of neuropathic pain.

博士
高雄醫學大學
醫學研究所
97
KMUP-1 is a xanthine derivate with soluble guanylyl cyclase activation, phosphodiesterases (PDE) inhibition and K+ channel (BKCa and KATP) opening activity in corpus cavernosum smooth muscle, basilar artery myocytes, aortic smooth muscle, and trachea. Our previous study shows that KMUP-1 inhibits tumor necrosis factor-alpha (TNF-α)-induced expression of iNOS in tracheal smooth muscle cells, involving the sGC/cGMP/PKG expression pathway. This study is aimed to investigate whether KMUP-1 has α1A/α1D-adremoceptor blocked activity in the prostate and anti-proliferation activity in benign and malignant prostate epithelial cells. Furthermore, we confirmed the effects of KMUP-1 on growth of LNCaP xenografts in nude mice. The results indicated that KMUP-1 possessed potent α1A/α1D-adrenoceptor binding inhibition activity, increased cAMP/cGMP levels and increased the expression of sGC, PKG and PKA protein in rat prostate. Moreover, KMUP-1 inhibited phenylephrine-induced ROCK2 expression. KMUP-1 inhibited cell growth, arrested the cell cycle at G0/G1 phase and increased the expression of p21 in PZ-HPV-7 cells. Treatment of LNCaP cells (androgen-dependent prostate cancer cell) with KMUP-1 resulted in cell cycle arrest and apoptotic activities. Moreover, KMUP-1 inhibited androgen receptor and prostate specific antigen (PSA) mRNA and protein expression. Regular administration of KMUP-1 suppressed the LNCaP xenograft tumor growth in nude mice. These results broaden our knowledge of sGC/cGMP/PKG and ROCK2 regulation on the relaxation and proliferation of prostate, which may help in the design of benign prostate hyperplasia (BPH) therapies that target these signaling pathways. Further, KMUP-1 might be used as a chemoprevention agent for preventing the development of prostate cancer without cardiovascular adverse effect.

碩士
高雄醫學大學
醫學系藥理學科碩士班
103
Atherosclerosis, a complex chronic inflammatory and metabolic disease, remains the leading cause of morbidity and mortality worldwide. Which is caused by the recruitment of blood monocytes, deposition of lipids, and formation of macrophage foam cells in the arterial wall. Apolipoprotein E-knockout (ApoE-KO) mice, a classic model of atherosclerosis, develop spontaneous atherosclerosis and thus have been widely used in cardiovascular research. In our previous studies, KMUP-1, a chemical synthetic xanthine-based derivative, has been shown its abilities of anti-inflammatory, anti-proliferation, and cardioprotective properties. This study aims to investigate whether KMUP-1 plays a protective role in inhibiting the progression of atherosclerosis in ApoE-KO mice. 　　For atherosclerosis studies, male ApoE-KO mice were randomly divided into four groups at the age of 8 weeks. High fat diet (HFD), prevention (KMUP-1-P) and treatment (KMUP-1-T) groups were fed with western diet containing 0.15% cholesterol and 21% fat (wt/wt) for 12 weeks. And the control (CTL) group was fed a normal chow diet for the same period. The KMUP-1-P group was administrated with KMUP-1 (5mg/kg/day) dissolved in drinking water for whole 12 weeks while the KMUP-1-T group for the last 4 weeks before sacrificing. According to our studies, KMUP-1 alleviated body and heart weight gain of ApoE-KO mice. In the echocardiography, FS% and EF% were improved in the KMUP-1-P and KMUP-1-T groups compared with the HFD group. The blood lipid profiles showed that KMUP-1 tended to decrease the level of TC, TG and LDL. However, KMUP-1 could somewhat increase the HDL level. Likewise, inflammatory cytokines IL-1?? and TNF-?? was decreased by KMUP-1. Histologically, atherosclerotic plaque with Oil-Red-O positive lesion areas were decreased in KMUP-1-given ApoE-KO mice. 　　KMUP-1 seemed to increase protein expressions of Atg7, Beclin-1, Bax and Bcl-2 in the aortic tissue of ApoE-KO. It is possible that KMUP-1 makes a protective role in the progression of atherosclerosis in ApoE-KO mice through the autophagy pathway. The results in our studies indicated that KMUP-1 may be a potential agent to restrict atherosclerosis development.

碩士
高雄醫學大學
醫學系藥理學科碩士班
103
Diabetes cardiomyopathy, a common disease occurring hypertension, hyperglycemia and ventricle hypertrophy in diabetes patients, is a major complication leads to heart failure. Recently, studies have reported that diabetes induced cardiomyocyte apoptosis and suppressed cardiac autophagy, showing that the relationship between the autophagy and apoptotic cell death pathways plays an important role in the pathogenesis of diabetic cardiomyopathy. Our recent studies indicated that KMUP-3 can induce autophagy in cardiomyocytes. Therefore, we further investigated whether KMUP-3’s promotion of autophagy activity can prevent high glucose (HG)-induce cardiac injury and improve cardiac functions in diabetes mellitus (DM) rats. In this study, we mimicked hyperglycemia condition in neonatal rat (1-3 day) cardiomyocytes with HG model. Cardiomyocytes were incubated in 30 mM HG in the presence or absence of KMUP-3 (1 to 10 &;#61549;M). An experimental diabetic rat model was induced by 65 mg/kg of streptozoticin (STZ). KMUP-3 was intraperitoneally injected at a dose of 1 mg/kg. Cardiac functions were evaluated by serial echocardiography. We found that KMUP-3 treatment attenuated HG-induced cell death by MTT assay. Additionally, KMUP-3 also inhibited HG-induced apoptosis, with associated increase of Bcl-2 protein, and decrease of Bax protein and caspase-3 cleavage. Microtubule-associated protein I light chain 3-II (LC3-II) is the key protein associated with autophagy. KMUP-3 significantly enhanced production of LC3-II and phosphorylation of AMPK in a time-dependent manner. As expected, KMUP-3 pretreatment dose-dependently reduced the HG-induced decrease of LC3-II, Atg7, and phosphor-AMPK expression. Fractional shortening (FS) and ejection fraction (EF), the index of left ventricular systolic function, were significantly decreased in DM group. Then compared with DM group, these changes were attenuated when diabetic rats were treated with KMUP-3 (P<0.05). In summary, KMUP-3 attenuates HG-induced cardiomyocytes apoptosis by inducing autophagy. These findings suggest that KMUP-3 may have great therapeutic potential in the treatment of diabetic cardiomyopathy.

碩士
高雄醫學大學
醫學系藥理學科碩士班
104
Vascular smooth muscle cells (VSMCs) apoptosis, occurring in many arterial diseases, including atherosclerosis, aneurysm formation and cell calcification, accelerates inflammation, smooth muscle cells loss and vascular remodeling. Abdominal aortic aneurysm (AAA), a chronic degenerative process of the abdominal aorta, is a life-threatening disease in the elderly population, especially in men. Pathologically, AAAs are associated with inflammation, smooth muscle cell apoptosis and matrix degradation. In our previous studies of high glucose-induced cardiomyocytes injury and diabetes rats’ model, we found that KMUP-3 can induce autophagy in cardiomyocytes. And reduce cardiac injury and improve cardiac functions. Vascular calcification is a sign of a degenerative inflammatory process involved in the arterial wall and it is a main risk factor in the cardiovascular field and may strengthen the rupture risk assessment of the AAA. We further investigated whether KMUP-3 can promote autophagy activity and attenuate VSMCs calcification, AAA formation and improve cardiac functions in apoE−/− mice. In this study, we used β-glycerophosphate (β-GP) to mimic the hyperphosphatemia condition which had already been proved to induce apoptosis in VSMCs. Primary 7-8 weeks male SD rats’ VSMCs were incubated in 10 mM β-GP in the presence or absence of KMUP-3 (0.1-10 μM) for 48 hr and 10 days. An experimental AAA model was induced via osmotic mini-pump (Alzet 2004) which was implanted into 24 weeks male apoE−/− C57BL/6 mice in order to administrate saline or angiotensin II (Ang II) s.c. at a dose of 1,000 ng/kg/min for 28 days, during which mice were fed with a Western diet (0.15% cholesterol and 21% milk fat). KMUP-3 was intraperitoneally injected at a dose of 1 mg/kg/day. At the end of the experimental period, we used transthoracic echocardiography to measure cardiac function, such as left ventricular end-systolic dimension (LVESD), LV end-diastolic dimension (LVEDD) and LV fractional shortening (%FS). We found that KMUP-3 treatment attenuated β-GP-induced cell death and VSMCs calcification by MTT assay, Alizarin Red S staining, Von Kossa staining, calcium deposition and ALP activity. Additionally, KMUP-3 also inhibited β-GP-induced apoptosis, with associated increased the activity of Bcl-2, and decreased the expression of Bax. The Annexin V/PI positive cell numbers in flow cytometry are also decreased. KMUP-3 significantly enhanced autophagy markers in VSMCs, such as LC3-II, ATG5 and phosphorylation of AMPK in pretreatment manner. In our animal model, KMUP-3 could not only reduce the formation of the aneurysm and decrease α-SMA depletion, vascular fibrosis, elastin degradation and calcium deposition at abdominal aorta, but also improve cardiac function of Ang II-infused APOE mice. The results in our studies indicated that KMUP-3 could dose-dependently attenuate β-GP-induced calcium deposition, ALP activity, the expression of angiogenesis, apoptosis and osteochondrogenic-related proteins through autophagy. Furthermore, KMUP-3 also has great capabilities to restrict AAA dilation. Based on our results, KMUP-3 may be a potent agent to decrease VSMCs calcification and AAA formation.

博士
高雄醫學大學
醫學研究所
99
The aim of this study was to investigate the inotropic effect and cardioprotective mechanism of KMUP-3 (7-[2-[4-(4-Nitro- benzene)-piperazinyl]-ethyl]-1,3-dimethyl-xanthine). KMUP-3 was a new drug synthesized in our laboratory and has been demonstrated to have phosphodiesterase inhibition, endothelial nitric oxide synthase (eNOS) enhancement, and KATP channel opening activities in human umbilical vein endothelial cells, aortic smooth muscle cells, and tracheal smooth muscle cells previously. It has been shown that cAMP and cGMP are involved in the cardiac inotropic effect, while KATP channel opening and eNOS activation have been shown to have cardioprotective effect. Here, we investigated the influence of KMUP-3 of cardiac output and protein expression in isolated atrium and Wistar rats in vivo. Our results demonstrated that Through cAMP enhancement, KMUP-3 increased contractility of isolated atrium and ventricule in Wistar rats. In isolated right atrium, KMUP-3 decreased the atrial contraction rate through activation of eNOS and parasympathetic nervous system. KMUP-3 increased the expression of PKA, eNOS, RhoA and Rho kinase (ROCK). The ROCK inhibitor, Y-27632 blocked the inotropic effect of KMUP-3, which suggested the involvement of RhoA/ROCK pathway in cardiac contractility. In the second part of the study, we further investigated the cardioprotective effect of KMUP-3 in myocardial infarction (MI) rats. Wistar rats were randomized into three groups: MI, MI + KMUP-3 group, and sham group. MI was induced by ligation of the left anterior descending coronary artery. After recovery, MI + KMUP-3 group received KMUP-3 (0.3 mg/kg/day) infusion for 4 weeks, while MI and sham group received vehicle only. To further confirm the eNOS-dependent activity, KMUP-3 was applied in the culture of transforming growth factor-β (TGF-β)-stimulated human cardiac fibroblasts (HCFs). KMUP-3 treatment attenuated cardiac hypertrophy with reduced infarction size after MI and improved cardiac output subsequently. The fibrotic area was reduced by KMUP-3 both in central, peri- and non-infarction area. KMUP-3 enhanced eNOS and tissue inhibitor of metalloproteinase-1 (TIMP-1) expression, with reduction of matrix metalloproteinase-9 (MMP-9) expression in MI rats. In HCFs, the ability of KMUP-3 in reducing MMP-9 and enhancing TIMP-1 expression was blocked by pretreatment with eNOS inhibitor Nω-nitro-L-arginine methyl ester (L-NAME). Thus, KATP channel opener KMUP-3 preserved cardiac function after MI through eNOS enhancement. KMUP-3 restored the myocardial MMP-9/TIMP-1 balance and attenuated ventricular remodeling with an eNOS-dependent mechanism.

碩士
高雄醫學大學
藥理學研究所
99
Osteoclasts are responsible for bone erosion in diseases as diverse as osteoporosis, periodonitis, and rheumatoid arthritis. Inhibition of osteoclast differentiation and bone resorption is considered as an effective therapeutic approach in the treatment of bone loss. Recently, phosphodiesterase (PDE) 4 inhibitors have been shown to have therapeutic effects in different experimental osteopenia models. The effect of KMUP-1, a PDE inhibitor, in M-CSF (macrophage colony-stimulating factor) and receptor activator of nuclear factor kappa B (RANKL)-induced osteoclast differentiation was examined in this study. In vitro, we found that KMUP-1 inhibited RANKL-mediated osteoclast differentiation in bone marrow macrophages (BMMs) of mice in a dose-dependent manner without any evidence of cytotoxicity and attenuated the bone resorption activity of differentiated osteoclasts. KMUP-1 also suppressed the MAP kinases, NF-κB, PI3K/Akt signaling pathways and the induction of c-fos and NFATc1 during osteoclastogenesis. We further investigated the effects of KMUP-1 on ovariectomy-induced bone loss using Micro-CT and serum markers assay for bone remodeling. Mice treated with KMUP-1 demonstrated attenuation of bone erosion based on Micro-CT and biochemical markers of bone turnover. These results collectively suggested that KMUP-1 demonstrated inhibitory effects on osteoclast differentiation in vitro, and suppressed ovariectomy-induced bone loss in vivo. Thus, KMUP-1 may serve as a therapeutic drug in the prevention of bone loss.

碩士
高雄醫學大學
藥理學研究所
98
In bone remodeling, an imbalance caused by increased bone resorption over bone formation leads to skeletal diseases such as osteoporosis, periodontitis, and rheumatoid arthritis. Recently, phosphodiesterase (PDE) 4 inhibitors have been shown to have therapeutic effects in different experimental osteopenia models. The receptor activator of nuclear factor-B ligand (RANKL) plays a critical role in the differentiation and bone resorptive activity of osteoclasts. Thus, in this study, we performed to determine whether KMUP-1, a PDE3, 4, 5 inhibitor, can attenuate the differentiation and proliferation of RANKL-induced osteoclast-like cells from RAW264.7 cells. In vitro, we found that KMUP-1 inhibited the RANKL-induced tartrate-resistance acid phosphatase (TRAP, a marker for osteoclast differentiation) activity and the formation of multinucleated osteoclasts in dose-dependent manner without any cytotoxicity. KMUP-1 also attenuated the bone resorption activity of mature osteoclasts. In addition, KMUP-1 inhibited inflammatory cytokines release and HMGB1 translocation which reported to play a major role in osteoclastogenesis. Furthermore, KMUP-1 prevented RANKL-induced activation of signaling molecules (Akt, MAP kinases and NF-κB) and key transcription factors (c-Fos and NFATc1) during early osteoclastogenesis. We also observed that KMUP-1 attenuated the activity and protein expression of matrix metalloproteinase-9 (MMP-9) and MMP-2. In vivo, Osteoarthritis was induced by intraarticular injection of monosodium iodoacetate (MIA) into knee joints of rats, KMUP-1 1, 2.5 and 5 mg/kg were orally 4 administered once a day for 7 days before and after MIA injection. We found that KMUP-1 was effective in articular cartilage erosion in MIA-induced osteoarthritis. Taken together, our results demonstrate that KMUP-1 potentially inhibits RANKL-induced osteoclast differentiation and proliferation by attenuating the downstream signaling molecules and transcription factors required for osteoclastogenesis in vitro. KMUP-1 attenuates MIA-induced osteoarthritis in vivo. Thus, we indicate that KMUP-1 may be a new therapeutic treatment for bone loss diseases.