肾气丸加减方对2型糖尿病小鼠胰岛β细胞PI3K/Akt/GSK-3β信号通路的影响  

Effects of Modified Shenqi Pill on pancreaticβcell PI3K/Akt/GSK-3βsignal pathways of type 2 diabetes mice

作　　者：张月颖 张珊 温志歌 倪青[1] ZHANG Yueying;ZHANG Shan;WEN Zhige;NI Qing(Guang’anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China)

机构地区：[1]中国中医科学院广安门医院,北京100053

出　　处：《中华中医药杂志》2024年第11期5875-5882,共8页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82174354)。

摘　　要：目的:探讨肾气丸加减方通过调控PI3K/A kt/G S K-3β信号通路保护胰岛β细胞功能的机制。方法:将C57BL/6小鼠随机分为空白组(n=10),模型组(n=12),肾气丸加减方低(n=11)、中(n=12)、高(n=12)剂量组,罗格列酮组(n=11)以及通路阻滞剂组(n=10)。观察小鼠体质量、外周糖脂代谢指标、胰岛素分泌水平,Tunel法测定胰岛β细胞凋亡水平,透射电子显微镜观察胰岛β细胞超微结构,Western blot法检测PI3K/Akt/GSK-3β信号通路以及PDX-1和MafA的蛋白表达水平。结果:干预8周后,与空白组比较,模型组小鼠体质量及空腹血糖水平显著升高(P<0.01),胰岛素分泌水平显著下降(P<0.01),胰岛β细胞凋亡率显著升高(P<0.01);胰岛β细胞PI3K、p-PI3K、Akt、p-Akt、p-GSK-3β、PDX-1以及MafA蛋白表达显著降低(P<0.05,P<0.01),GSK-3β蛋白表达量显著升高(P<0.01),电子显微镜下线粒体嵴结构消失。干预8周后,与模型组比较,肾气丸加减方高剂量组与罗格列酮组体质量及空腹血糖水平显著降低(P<0.01),胰岛素分泌水平显著升高(P<0.05),β细胞凋亡率显著降低(P<0.01),肾气丸加减方中、高剂量组PI3K、p-PI3K、Akt、p-Akt、p-GSK-3β、PDX-1以及MafA蛋白表达量均显著升高(P<0.05,P<0.01),GSK-3β蛋白表达量显著降低(P<0.01),加入通路阻滞剂LY294002后其作用被抵消(P<0.05,P<0.01),电子显微镜下线粒体形态规则,嵴结构存在。结论:肾气丸加减方可作用PI3K/Akt/GSK-3β信号通路改善T2DM小鼠糖脂代谢水平,保护线粒体嵴结构以减少氧化应激对胰岛β细胞的损伤。Objective:To explore the mechanism of modified Shenqi Pill in protecting pancreaticβ-cell function by regulating PI3K/Akt/GSK-3βsignaling pathway.Methods:C57BL/6 mice were randomly divided into blank group(n=10),model group(n=12),low-dose group(n=11),medium-dose group(n=12),high-dose group(n=12)of Modified Shenqi Pill,rosiglitazone group(n=11),and pathway blocker group(n=10).The body weight,peripheral glucose and lipid metabolism indexes,and insulin secretion levels of the mice were observed.The apoptosis level of pancreaticβcells was determined by Tunel method,the ultrastructure of pancreaticβcells was observed by transmission electron microscopy,and the protein expression of PI3K/Akt/GSK-3βsignaling pathway and PDX-1 and MafA were detected by Western blot.Results:After 8 weeks of treatment,compared with the blank group,the body weight and fasting blood glucose levels of mice in the model group were significantly increased(P<0.01),the insulin secretion level was significantly decreased(P<0.01),and the apoptosis rate of pancreaticβcells was significantly increased(P<0.01).The protein expressions of PI3K,p-PI3K,Akt,p-Akt,p-GSK-3β,PDX-1 and MafA of pancreaticβcells in the model group were decreased(P<0.05,P<0.01),the protein expression of GSK-3βwas significantly increased(P<0.01),and the mitochondrial cristae structure disappeared under electron microscopy.After 8 weeks of treatment,compared with the model group,the body weight and the fasting blood glucose level of the high-dose Modified Shenqi Pill and rosiglitazone groups were significantly reduced(P<0.01),the insulin secretion level was significantly increased(P<0.05),theβ-cell apoptosis rate was significantly reduced(P<0.01),the protein expression levels of PI3K,p-PI3K,Akt,p-Akt,p-GSK-3β,PDX-1 and MafA in the medium and high-dose Modified Shenqi Pill groups were significantly increased(P<0.05,P<0.01),the protein expression level of GSK-3βwas significantly decreased(P<0.01),and its effect was offset by the addition of pathway blocker LY294002(P<0.05

关 键 词：肾气丸加减方 2型糖尿病 胰岛Β细胞 PI3K/Akt/GSK-3β PDX-1/MafA 

分 类 号：R285.5[医药卫生—中药学]