發(fā)布時(shí)間：2018-04-15 20:53

  本文選題：過(guò)氧化物酶體增殖物激活受體γ + 噻唑烷二酮類(lèi)��； 參考：《大連醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：眾所周知，吡格列酮屬于TZD類(lèi)（thiazolidinediones，噻唑烷二酮類(lèi)）降血糖藥物。屬于PPARγ（Peroxisome Proliferator-activated Receptor-γ，過(guò)氧化物酶體增殖物激活受體γ）激動(dòng)劑。通過(guò)改善外周及肝臟對(duì)胰島素的敏感性來(lái)控制血糖水平。自1999年上市以來(lái)廣泛應(yīng)用于治療2型糖尿病。但近期有資料提示吡格列酮有增加膀胱癌風(fēng)險(xiǎn)的可能。吡格列酮屬于核受體超家族成員，通過(guò)與同源維甲酸受體X形成異二聚體，影響基因轉(zhuǎn)錄、蛋白合成，調(diào)節(jié)脂質(zhì)代謝、細(xì)胞生長(zhǎng)及分化等。噻唑烷二酮類(lèi)藥物中，曲格列酮1997年上市，2000年因其肝衰竭風(fēng)險(xiǎn)撤市。FDA（Food and Drug Admistraton，歐洲藥品管理局）暫停了另一種TZD類(lèi)藥物羅格列酮的銷(xiāo)售，因其可增加心血管事件的風(fēng)險(xiǎn)。目前，吡格列酮是唯一廣泛使用中的TZD類(lèi)藥物。因此，評(píng)估吡格列酮與膀胱癌的風(fēng)險(xiǎn)是至關(guān)重要的，本研究探討吡格列酮對(duì)膀胱正常移行上皮細(xì)胞及膀胱癌細(xì)胞生長(zhǎng)的影響，旨在為臨床用藥的安全性提供依據(jù)。 方法：原代培養(yǎng)大鼠膀胱正常移行上皮細(xì)胞。將膀胱正常移行上皮細(xì)胞及膀胱癌細(xì)胞株(J82)，分別以0μmol/L、5μmol/L、10μmol/L、20μmol/L、40μmol/L濃度吡格列酮作用0h、24h、48h、72h、8d、10d，電子顯微鏡觀察細(xì)胞形態(tài)及生長(zhǎng)情況；MTT（3-（4,5-Dimethylthiazol-2-y）2,5-diphenyltetrazolium bromide，噻唑藍(lán)）法檢測(cè)吡格列酮對(duì)細(xì)胞生長(zhǎng)的抑制作用；流式細(xì)胞技術(shù)分析細(xì)胞凋亡情況；實(shí)時(shí)定量PCR（Polymerase Chain Reaction，聚合酶鏈?zhǔn)椒磻?yīng)）法檢測(cè)腫瘤相關(guān)基因CyclinD1、P53及凋亡相關(guān)基因Bcl-2、Bax的表達(dá)；Western blot（蛋白印跡技術(shù)）檢測(cè)Cyclin D1、P53、Bcl-2、Bax蛋白的表達(dá)。最后，依據(jù)實(shí)驗(yàn)結(jié)果分析吡格列酮對(duì)膀胱正常移行上皮細(xì)胞及膀胱癌細(xì)胞生長(zhǎng)的影響。 結(jié)果：吡格列酮作用于膀胱正常移行上皮細(xì)胞48h后出現(xiàn)細(xì)胞凋亡，，作用強(qiáng)度呈濃度-時(shí)間依賴(lài)型。膀胱癌細(xì)胞J82則未見(jiàn)促細(xì)胞增殖或抑制現(xiàn)象。10μmol/L吡格列酮處理膀胱正常移行上皮細(xì)胞24h、72h的凋亡率（vs對(duì)照組）分別為：24h：18.8％vs9.4％、72h：49.7％vs11.3％。相同劑量吡格列酮處理大鼠肝臟正常細(xì)胞、腎臟正常細(xì)胞72h，均未見(jiàn)細(xì)胞凋亡現(xiàn)象。實(shí)時(shí)定量PCR法檢測(cè)10μmol/L吡格列酮作用于膀胱正常移行上皮細(xì)胞及J82細(xì)胞24h、48h、72h后腫瘤相關(guān)基因Cyclin D1、P53及凋亡相關(guān)基因Bcl-2、Bax的表達(dá)均未見(jiàn)差異。Western blot法檢測(cè)10μmol/L吡格列酮作用后Cyclin D1、P53、Bcl-2、Bax蛋白的表達(dá)，24h、48h、72h時(shí)Cyclin D1、P53、Bcl-2、Bax均未見(jiàn)差異。長(zhǎng)期作用膀胱正常移行上皮細(xì)胞也未見(jiàn)明顯差異。但J82細(xì)胞中作用8d后Cyclin D1、P53蛋白表達(dá)減弱。 結(jié)論：吡格列酮不增加膀胱正常移行上皮細(xì)胞癌變趨勢(shì)，亦不促進(jìn)膀胱癌細(xì)胞增殖；長(zhǎng)期培養(yǎng)（8d）可一定程度抑制膀胱癌細(xì)胞活性；吡格列酮促進(jìn)膀胱正常移行上皮細(xì)胞凋亡，具有組織特異性。從現(xiàn)有結(jié)果推測(cè)：糖尿病患者應(yīng)用吡格列酮可能并不增加其罹患膀胱癌風(fēng)險(xiǎn)；糖尿病已合并膀胱癌的患者使用吡格列酮的治療中，不會(huì)促進(jìn)腫瘤細(xì)胞的生長(zhǎng)，可能還會(huì)在一定程度上抑制腫瘤的發(fā)展。
[Abstract]:Objective: as everyone knows, pioglitazone belongs to the TZD class (thiazolidinediones, thiazolidine ketone two) hypoglycemic drugs. PPAR (Peroxisome Proliferator-activated Receptor- belongs to gamma gamma, peroxisome proliferator activated receptor gamma agonist). By improving peripheral and hepatic insulin sensitivity to control blood sugar levels. Since 1999 the market is widely used in the treatment of type 2 diabetes. But the recent data suggest that pioglitazone may increase the risk of bladder cancer. Pioglitazone belongs to the nuclear receptor superfamily, through the formation of two dimers and homology of retinoic acid receptor X, gene transcription, protein synthesis, regulation of lipid metabolism, cell growth and differentiation. Two thiazolidinediones troglitazone, listed in 1997 in 2000, because of the risk of liver failure,.FDA (Food and Drug from Admistraton, the European Medicines Agency) suspended another TZD drugs Compound rosiglitazone sales, because it can increase the risk of cardiovascular events. At present, pioglitazone is TZD drugs widely used in risk assessment. Therefore, the bladder cancer is very important, this study was to investigate the effect of pioglitazone in normal prostate epithelial cells and the growth of bladder cancer cells of the bladder, in order to provide a basis for the safety of clinical medication.
Methods: primary cultured rat bladder transitional epithelial cells. Normal bladder transitional epithelial cells and normal human bladder cancer cell line (J82), respectively, 0 mol/L, 5 mol/L, 10 mol/L, 20 mol/L, 40 mol/L 0h 24h, pioglitazone, 48h, 72h, 8D. 10d, electron microscope observe the cell morphology and growth; MTT (3- (4,5-Dimethylthiazol-2-y) 2,5-diphenyltetrazolium bromide, MTT) method was used to detect the inhibitory effect of pioglitazone on cell growth, cell apoptosis analysis; flow cytometry; real-time quantitative PCR (Polymerase Chain Reaction, polymerase chain reaction) method for the detection of tumor related gene CyclinD1, P53 and apoptosis related gene Bcl-2, expression of Bax; Western blot (Western blot) detection of Cyclin D1, P53, Bcl-2, Bax protein expression. Finally, based on the analysis of experimental results of pioglitazone on normal bladder transitional epithelial cells and bladder The effect of cystocysteine cell growth.
Results: the appearance of apoptosis of pioglitazone effect on normal bladder transitional epithelial cells after 48h intensity with concentration time dependent. The bladder cancer cell J82 was found in cell proliferation or inhibit the phenomenon of.10 mol/L pioglitazone treatment of normal bladder transitional epithelial cells 24h, 72h apoptosis rate (vs control group) respectively. 24h:18.8%vs9.4% 72h:49.7%vs11.3%., the same dose of pioglitazone treated rats in normal liver cells, normal kidney cell 72h, there were no cell apoptosis. Quantitative real-time PCR to detect 10 mol/L pioglitazone effect on normal bladder transitional epithelial cells and J82 cells in 24h, 48h, 72h of tumor related gene Cyclin and apoptosis related gene D1, P53 Bcl-2, the expression of Bax showed no difference between.Western detection method of blot 10 mol/L Cyclin D1 P53 after the effect of pioglitazone, Bcl-2, and Bax protein expression, 24h, 48h, 72h Cyclin D1, P53, Bcl There was no difference between -2 and Bax. There was no significant difference in the normal transitional epithelial cells of the bladder in the long term, but the expression of P53 protein was weakened after the action of Cyclin D1 in J82 cells.
Conclusion: pioglitazone does not increase the normal bladder transitional epithelial cells nor canceration trend, promote the proliferation of bladder cancer cells; long term culture (8D) can inhibit bladder cancer cell activity; pioglitazone promotes normal bladder transitional epithelial cell apoptosis, tissue specific. Inferred from existing results: diabetic patients with pioglitazone may not increase its the risk of bladder cancer risk; diabetes mellitus has combined with bladder cancer patients with the use of pioglitazone in the treatment does not promote tumor cell growth, may inhibit tumor development in a certain extent.

【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R737.14


本文編號(hào)：1755733