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目的：探讨高浓度葡萄糖对体外培养大鼠胰岛细胞凋亡的影响及作用机制。方法：SD 大鼠胰岛细胞原代培养，不同浓度的葡萄糖处理后，采用MTT 比色法检测细胞存活率，Hoechst-PI 染色观察细胞凋亡，电镜观察细胞超微结构改变，RT-PCR 方法检测Bax 及Bcl-2 相关基因mRNA 表达水平。结果：5.5 mmol·L-1，11.1 mmol·L-1和22.2 mmol·L-1葡萄糖处理后，胰岛细胞活性分别下降到78.08 %± 2.29 %、58.39 %± 3.13 %和36.05 %± 2.63 %，与阴性对照组比较差异具有统计学意义(P<0.05)；Hoechst-PI染色结果显示随着葡萄糖作用浓度的增加，凋亡细胞数量也增加；RT-PCR 显示胰岛细胞经不同浓度葡萄糖处理后，Bax mRNA的表达明显上调，Bcl-2 mRNA表达下调(P<0.05)；电镜观察结果显示随着葡萄糖作用浓度的增加，胰岛细胞超微结构损害程度依次加重。结论：高浓度葡萄糖能明显引起胰岛细胞活性的下降，诱导凋亡反应的发生，凋亡机制与Bcl-2 家族蛋白相关。

英文摘要:

Objective:To investigate the effect and mechanismof high concentration of glucose on apoptosis in cultured rat islets.Methods:SD rats islet cells were primarily cultured, treated with different concentrations of glucose, and the cells viability were detected by MTT colorimetric, and the cells apoptosis were checked by Hoechst-PI staining, and the cell ultrastructural changes were detected by transmission electron microscopy, and the expression level of apoptosis-related genes were detected by RT-PCR.Results:After treated with glucose with the different dose of 5.5 mmol·L-1, 11.1 mmol·L-1 and 22.2 mmol·L-1, islet cells activity was decreased to 78.08 %± 2.29 %, 58.39 %± 3.13 % and 36.05 %± 2.63 %, which had statistical significance compared with control group (P<0.05); Hoechst-PI results showed that apoptosis cells were increased companied with the elevation of concentration of glucose; the results of RT-PCR detection showed that Bax mRNA level was upregulated and Bcl-2 mRNA level was downregulated with the elevation of glucose concentration; and the results of electromicroscopy showed that the ultrastructure of islets cells also was changed with the concentration of glucose.Conclusion:High glucose obviously can cause the decline of cell activity in islet cells, and induce the apoptosis reaction, and the apoptotic mechanismmay be related to Bcl-2 family proteins.

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