罗格列酮对RIN-m细胞高糖损害干预作用研究

The effect of rosiglitazone on high glucose-induced RIN-m cells impairment

目的研究罗格列酮(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。

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 μmol/L （ NG ＋ R）, high glucose （33.3 mmol/L, HG） and HG ＋ R 50 μmol/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 - PER 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 13 cells against the glucotoxicity.