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目的观察重组胰生定多肽(EXf)对H2O2诱导的INS-1胰岛β细胞凋亡的影响。方法将INS-1细胞分成5组:空白对照组、阴性对照组、阳性对照组(EX-4)、给药组(EXf,终浓度分别为10、100 nmol/L),每组至少6孔。除空白对照组外,均给予终浓度为100μmol/L H2O2诱导INS-1胰岛β细胞凋亡模型。通过MTT检测法、Hoechest 33342染色法、DNA ladder检测法,观察EXf对H2O2诱导INS-1胰岛β细胞细胞存活率、形态学变化和凋亡的影响。结果 MTT实验结果显示与模型对照组相比,EXf能使H2O2处理的INS-1胰岛β细胞存活率明显增加(P<0.01),并呈剂量依赖性。Hoechst 33342染色检测显示给药组细胞凋亡数量明显减少,正常细胞数量明显增加。EXf能明显抑制DNA ladder的出现,且呈一定量效关系。结论 EXf对H2O2诱导的INS-1胰岛β细胞的凋亡具有明显的保护作用。
Objective To investigate the effect of EXf on H2O2-induced pancreatic β-cell apoptosis in INS-1 cells. Methods INS-1 cells were divided into 5 groups: blank control group, negative control group, positive control group (EX-4), administration group (EXf, final concentration of 10,100 nmol / L) . Except the blank control group, the model of INS-1 islet β cell apoptosis was induced by H2O2 at a concentration of 100 μmol / L. The effect of EXf on the cell viability, morphological changes and apoptosis of INS-1 islet β cells induced by H2O2 was observed by MTT assay, Hoechest 33342 staining and DNA ladder assay. Results The results of MTT showed that compared with the model control group, the survival rate of INS-1 cells induced by H2O2 was significantly increased (P <0.01) by EXf in a dose-dependent manner. Hoechst 33342 staining showed that the number of apoptotic cells in the treated group was significantly reduced and the number of normal cells was significantly increased. EXf can significantly inhibit the appearance of DNA ladder, and showed a dose-effect relationship. Conclusion EXf has obvious protective effect on the apoptosis of INS-1 islet β cells induced by H2O2.