罗格列酮对rinm细胞高糖损害干预作用分析.pdf

罗格列酮对RIN-m 细胞高糖损害干预作用分析 【摘要】研究罗格列酮(rosiglitazone，RSG)对胰岛β 细胞高糖损害的干预作 用。方法 采用生理浓度葡萄糖（5.5 mmol/L）、生理浓度葡萄糖+罗格列酮、 高浓度葡萄糖（33.3 mmol/L）、高浓度葡萄糖+罗格列酮培养RIN-m 细胞。以 放射免疫法检测胰岛素分泌水平。以流式细胞仪及TUNEL 法检测RIN-m 细胞凋 亡。RT-PCR 方法检测胰腺十二指肠同源盒-1 （pancreatic duodenal homeobox factor-1，PDX-1）和胰岛素mRNA 表达。结果 ①RIN-m 细胞与33.3 mmol/L 的 葡萄糖孵育2 周后胰岛素分泌功能开始下降，细胞凋亡率增长2.7 倍（P＜ 0.01）。而罗格列酮可以修复胰岛素的分泌，降低RIN-m 细胞凋亡率。②罗格 列酮上调 PDX-1 （1.30±0.06 vs. 1.61±0.10，P＜0.01）和胰岛素 (1.75± 0.09 vs. 1.98±0.11，P＜0.01) mRNA 表达。结论 高糖抑制胰岛β 细胞胰岛 素分泌，并诱导其凋亡。罗格列酮具有直接保护β 细胞免于高糖毒性的作用。 【关键词】罗格列酮 RIN-m 细胞 凋亡 胰腺十二指肠同源盒-1 Abstract: Objective To investigate the effect of rosiglitazone (R) on high glucose-induced RIN-m cell impairment. Methods The RIN-m cells were cultured in media containing normal glucose (5.5 mmol/L， NG)， NG+R 50 μ mol/L (NG+R)， high glucose (33.3 mmol/L， HG) and HG+R 50 μ mol/L (HG+R). Insulin secretion was measured by radioimmunoassay. Apoptosis of RIN-m cells were determined by in situ TUNEL method combined with flow cytometry. The mRNA expression of pancreatic duodenal homeobox factor-1 (PDX-1) and insulin was measured by semi-quantitative RT-PCR assay. Results (1) After 2 weeks of incubation， the presence of high glucose increased the apoptosis of RIN-m cells to 2.7 folds and decreased the insulin secretion (P＜ 0.05) . The addition of R inhibited the high glucose-induced increase of apoptosis of RIN-m cells and improved the insulin secretion (P＜ 0.05). (2) Incubation of RIN-m cells with high glucose plus R for 2 weeks， increased the mRNA expression of PDX-1 ［from （1.30±0.06 ） to (1.63±0.10)，P＜0.05］ and that of insulin ［from (1.75±0.09) to ( 1.98±0.11)， P＜0.05］. Conclusions High glucose can induce cell apoptosis and decrease insulin secretion. R (rosiglitazone) has direct protective effects on β cells against the glucotoxicity. Key words: rosiglitazone; RIN-m cell; apoptosis; pancreatic duodenal homeobox factor-1 （PDX-1） 持续高糖环境导致胰岛功能缺陷和胰岛数量的减少［1］