半夏泻心汤对过氧化氢叔丁醇诱导的MIN6细胞凋亡及PI3K/AKT/FOXO1信号通路的影响

Effects of Banxia Xiexin Decoction (半夏泻心汤) on TBHP-induced MIN6 Cell Apoptosis and PI3K/AKT/FOXO1 Signaling Pathway

目的探讨半夏泻心汤治疗2型糖尿病的可能作用机制。方法MIN6细胞分为空白组、模型组、利拉鲁肽组及半夏泻心汤低、中、高剂量组,半夏泻心汤低、中、高剂量组分别加入0.25、0.5、1.0mg/ml浓度的半夏泻心汤lOjil预预处理12h,利拉鲁肽组加入0.lS^g/ml利拉鲁肽注射液10|il处理12h。半夏泻心汤各剂量组、利拉鲁肽组和模型组用过氧化氢叔丁醇(TBHP)孵育4h建立细胞凋亡模型,MTT法检测各组细胞的凋亡抑制率,Western blot法检测各组细胞中蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)、叉头转录因子1(FOXO1)、磷酸化叉头转录因子1(p-FOXOl)及P27蛋白的表达。结果与空白组比较,模型组对MIN6细胞凋亡的抑制率明显升高(P<0.05);与模型组比较,利拉鲁肽组及半夏泻心汤中、高剂量组凋亡抑制率均降低(P<0.05);各干预组间比较差异无统计学意义(P>0.05)。与空白组比较,模型组P-AKT/AKT、p-FOXOl/FOXOl表达明显下调,P27表达明显上调(P<0.05);与模型组比较,除半夏泻心汤低剂量组p-FOXOl/FOXOl外,利拉鲁肽组及半夏泻心汤各剂量组p-AKT/AKT.p-FOXOl/FOXOl表达均明显上调,P27表达均明显下调(P<0.05);与半夏泻心汤高剂量组比较,利拉鲁肽组、半夏泻心汤中剂量组p-FOXOl/FOXOl表达明显上调(P<0.05);与半夏泻心汤低剂量组比较,半夏泻心汤中、高剂量组及利拉鲁肽组P27表达均明显下调(P<0.05)o结论半夏泻心汤可显著抑制MIN6胰岛B细胞的凋亡而发挥防治2型糖尿病的治疗作用,以髙剂量效果更佳,其机制可能是通过激活PI3K/AKT/FOXO1信号通路,从而调节相关蛋白表达。

Objective To investigate the possible mechanism of type 2 diabetes treated by Banxia Xiexin Decoction(半夏泻心汤).Methods MIN6 cells were divided into a blank group,a model group,a liraglutide group,and Banxia Xiexin Decoction low dose,medium dose and high dose group.Banxia Xiexin Decoction low dose,medium dose and high dose group were respectively added 10 pel of Banxia Xiexin Decoction at concentration of 0.25,0.5,1.0mg/ml pretreated for 12h,and 0.18 jxg/ml Liraglutide Injection was added to the liraglutide group for 12h.All groups of Banxia Xiexin Decoction,the liraglutide group and the model group used tert-butanol hydrogen peroxide(TBHP)to establish cell apoptosis model.MTT method was used to detect the cell apoptosis rate of each group,and Western blot method was used to detect the expression of cellular protein kinase B(AKT),phosphorylated protein kinase B(p-AKT),forkhead transcription factor 1(FOXOl),phosphorylated forkhead transcription factor 1(p-FOXOl)and P27 protein.Results Compared with the blank group,the inhibition rate of MIN6 cells in the model group was significantly increased(P<0.05).Compared with the model group,the inhibition rates of the liraglutide group and the Banxia Xiexin Decoction medium dose and high dose group were all reduced(P<0.05).There was no significant difference between the intervention groups(P>0.05).Compared with the blank group,the expression of p-AKT/AKT,p-FOXOl/FOXOl in the model group was significantly down-regulated,and the expression of P27 was significantly up-regulated(P<0.05).Compared with the model group,except p-FOXOl/FOXOl in the Banxia Xiexin Decoction low dose group,the expression of p-AKT/AKT,p-FOXl/FOXl in the liraglutide group and different dose of Banxia Xiexin Decoction groups were significantly upregulated,and the expression of P27 was significantly down-regulated(P<0.05).Compared with the Banxia Xiexin Decoction high-dose group,the expression of p-FOXl/FOXl in the liraglutide group and the Banxia Xiexin Decoction medium-dose group was significantly upregulated(P<0.05).Compared with the Banxia Xiexin Decoction low-dose group,the expression of P27 in Banxia Xiexin Decoction medium-dose and high-dose group,and the liraglutide group was significantly down-regulated(P<0.05).Conclusion The medium and high dose of Banxia Xiexin Decoction can significantly inhibit the apoptosis of MIN6 islet 0 cells to treat type 2 diabetes,and it is more effective at high doses.The mechanism may be activating the PI3K/AKT/FOXO1 signaling pathway to regulate the expression of related proteins.