葛根素对大鼠星形胶质细胞增殖、凋亡和β-catenin、cyclinD1、Bax蛋白表达的影响

Effects of puerarin on proliferation,apoptosis,and expressions of β-catenin,cyclinD1,Bax proteins of rat astrocytes

目的:探讨葛根素对H_2O_2诱导的大鼠星形胶质细胞(AST)增殖、凋亡的影响及作用机制。方法:采用MTT法检测100、200、400、600、800、1 000μmol/L H_2O_2对AST活力的影响;将AST分为空白对照组(细胞未经任何处理)、H_2O_2组(400μmol/L H_2O_2处理AST 24 h)和葛根素+H_2O_2组(1 mmol/L葛根素处理30 min后,400μmol/L H_2O_2处理24 h),应用MTT、Annexin V-FITC/PI双染法及Western blot分别检测细胞活力、凋亡率及β-catenin、cyclin D1和Bax蛋白的表达。结果:随着H_2O_2浓度增加,AST活力降低(P<0.05)。与空白对照组比较,H_2O_2组细胞活力明显降低,凋亡率升高,β-catenin和cyclin D1表达降低,Bax表达升高(P<0.05);与H_2O_2组比较,葛根素+H_2O_2组细胞活力升高,凋亡率降低,β-catenin和cyclin D1表达升高,Bax表达降低(P<0.05)。结论:葛根素可减轻H_2O_2诱导的大鼠AST氧化应激损伤,机制与激活Wnt/β-catenin信号通路有关。

Aim:To investigate the effects and mechanism of puerarin on proliferation and apoptosis of rat astrocytes induced by H 2O 2.Methods:The effects of different concentrations(100,200,400,600,800,1 000μmol/L)of H2O2 on the viability of astrocytes were detected by MTT assay.Rat astrocytes were divided into blank control group(untreated cells),H2O2 group(treated with 400μmol/L H2O2 for 24 hours),puerarin+H2O2 group(treated with 1 mmol/L puerarin for 30 min and 400μmol/L H2O2 for 24 hours).MTT,flow cytometry and Western blot were used to detect cell viability,apoptosis and expressions ofβ-catenin,cyclinD1 and Bax proteins,respectively.Results:With the increase of H2O2 concentration,the activity of rat astrocytes decreased(P<0.05).Compared with the blank control group,the cell viability in the H2O2 group was significantly decreased,the apoptosis rate was significantly increased,the expressions ofβ-catenin and cyclinD1 were significantly decreased,and the expression of Bax was significantly increased(P<0.05).Compared with the H2O2 group,the cell viability in the puerarin+H2O2 group was significantly increased,the apoptosis rate was significantly decreased,the expressions ofβ-catenin and cyclinD1 were significantly increased,and the expression of Bax was significantly decreased(P<0.05).Conclusion:Puerarin can promote the proliferation and inhibit apoptosis of rat astrocytes induced by H 2O 2,and the mechanism is related to the activation of Wnt/β-catenin signaling pathway.