升清降糖合剂对H_2O_2诱导凋亡的胰岛β细胞周期和Caspase-3的影响

EFFECTS OF SHENGQING JIANGTANG MIXTURE ON CYCLE AND CASPASE-3 EXPRESSION OF PANCREATIC BETA-CELL APOPTOSIS INDUCED BY H_2O_2

目的:探讨升清降糖合剂(以下简称SQ)对氧化损伤的胰岛β细胞凋亡的保护作用。方法:以H2O2诱导建立胰岛细胞凋亡模型,采用半仿生提取法提取复方有效成分,以10μg/ml作用于胰岛细胞凋亡模型,采用TUNEL法检测细胞凋亡,流式细胞术检测细胞周期,分光光度法检测凋亡蛋白Caspase-3的活性。结果:H2O2可诱导胰岛β细胞凋亡,加用SQ后细胞凋亡现象明显下降。与正常组比较,模型组RINm5F细胞在G2/M期比例减少(P<0.01),保护组RINm5F细胞在G0/G1的比例减少(P<0.05),在G2/M期细胞比例上升(P<0.01)。模型组RINm5F细胞Caspase-3活化增强,模型组原代胰岛细胞Caspase-3活化增强,保护组Caspase-3活性下降,与模型组比较,有统计学差异(P<0.01)。结论:SQ对氧化损伤诱导的胰岛β细胞凋亡具有保护作用,其机制与调节胰岛β细胞的细胞周期和调节凋亡相关蛋白Caspase-3表达有关。

Objective: To explore the mechanism of protective effect of shengqing jiangtang mixture on pancreatic β-cell apoptosis caused by oxidative injury through observing the effects of the mixture on cycle and Caspase-3 activity in apoptosis pancreatic β-cell induced by H2O2. Methods: Models of pancreatic β-cell apoptosis were induced by H2O2,and intervened with 10μg/ml shengqing jiangtang mixture, apoptosis of pancreatic β-cell was determined by TUNEL method, cell cycle was observed by flow cytometry, the Caspase-3 activity was determined by spectrophotometry method. Results: H2O2 could induce the apoptosis of primary pancreatic β-cell and RINm5F apoptosis, shengqing jiangtang mixture could reduce the apoptosis of pancreatic β-cell. The number of RINm5Fcells at G2/M phase was less than that of normal, shengqing jiangtang mixture could decrease the number at G0/G1 phase and increase the number at G2/M phase. The Caspase-3 activity was enhanced obviously in model cells , shengqing jiangtang mixture could lower Caspase-3 activity, there was significant difference between the model and mixture group. Conclusion: Shengqing jiangtang mixture can protect pancreatic β-cell from apoptosis caused by oxidative injury, which is related to regulating pancreatic β-cell cycle and apoptosis-related protein Caspase-3.