津力达颗粒对高糖作用下小鼠胰岛β细胞增殖和凋亡的影响

Effects of Jinlida Granules on Proliferation and Apoptosis of Mouse Islet β Cells Under High Glucose Environment

目的探讨津力达颗粒对高糖作用下小鼠胰岛β细胞增殖和凋亡的影响。方法将MIN6细胞分为正常对照(NC)组、津力达(JLD)组、高糖(HG)组和高糖+津力达(HG+JLD)组。培养48 h后,分别用2.8,16.7 mmol·L^(-1)葡萄糖KRBH缓冲培养液孵育0.5 h,酶联免疫吸附实验(ELISA)检测上清液胰岛素含量。采用噻唑蓝(MTT)法检测细胞增殖率,流式细胞仪检测细胞凋亡率,免疫印迹法(Western blotting)检测Caspase-3、Bax、Bcl-2和Smad2/3蛋白表达。结果与2.8 mmol·L^(-1)葡萄糖刺激比较,16.7 mmol·L^(-1)葡萄糖刺激时各组胰岛素分泌(GSIS)均升高(均P<0.01)。16.7 mmol·L^(-1)葡萄糖刺激时HG+JLD组GSIS高于HG组(P<0.01)。与HG组比较,HG+JLD组细胞增殖率增高(P<0.01),早晚期凋亡率降低(均P<0.01),Caspase-3-2、Bax蛋白表达下调(P<0.01或P<0.05),Bcl-2蛋白表达上调(P<0.01),p-Smad2/3蛋白表达下调(P<0.01)。结论津力达颗粒可改善高糖环境下的小鼠胰岛β细胞胰岛素分泌,促进细胞增殖,调节凋亡蛋白的表达,抑制细胞凋亡,其作用可能与抑制Smad2/3磷酸化有关。

Objective To investigate the effects of Jinlida granules on the proliferation and apoptosis of pancreatic isletβcells in mice induced by high glucose.Methods MIN6 cells were divided into normal control(NC)group,Jinlida(JLD)group,high glucose(HG)group and high glucose and JLD(HG+JLD)group.After culture for 48 h,the cells were treated with KRBH medium for 0.5 h as glucose stimulation(2.8 or 16.7 mmol·L^(-1)).The insulin content of supernatant was detected by ELISA;cell proliferation rate was detected by MTT assay;cell apoptosis rate was detected by flow cytometry;and expression of Caspase-3,Bax,Bcl-2 and Smad2/3 protein were detected by Western blotting.Results Compared with 2.8 mmol·L^(-1) glucose stimulation,the glucose-stimulated insulin secretion(GSIS)of each group increased at 16.7 mmol·L^(-1) glucose stimulation(all P<0.01).The GSIS of HG+JLD group was higher than that of HG group at 16.7 mmol·L^(-1) glucose stimulation(P<0.01).Compared with HG group,HG+JLD group had higher proliferation rate(P<0.01),lower in early and late apoptosis rate(all P<0.01),down-regulated Caspase-3-2,Bax protein expression(P<0.01,P<0.05),up-regulated Bcl-2 protein expression(P<0.01)and down-regulated p-Smad2/3 protein expression(P<0.01).Conclusion Under high glucose environment,Jinlida granules can improve insulin secretion,promote the proliferation,regulate apoptosis protein expression,and inhibit apoptosis of mouse pancreaticβcells,which may be related to inhibition of Smad2/3 phosphorylation.