白虎加人参汤加减方对2型糖尿病大鼠骨骼肌GLUT4表达及抗氧化应激作用机制的研究  

Study on the Modified Baihu Plus Renshen Decoction on the GLUT4 Expression in Skeletal Muscle and Mechanism of Anti-oxidative Stress of Type 2 Diabetes Mellitus Rats

作　　者：王芳 刘清[2] 高天慧 宋小妹[2] WANG Fang;LIU Qing;GAO Tianhui;SONG Xiaomei(Shaanxi Energy Institute,Xianyang 712000 Shaanxi,China;Shaanxi University of Chinese Medicine,Xianyang 712046 Shaanxi,China)

机构地区：[1]陕西能源职业技术学院,陕西咸阳712000 [2]陕西中医药大学,陕西咸阳712046

出　　处：《中药新药与临床药理》2023年第9期1203-1210,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：陕西省科技厅自然科学基础研究计划项目(2021IQ-888);陕西省教育厅科研计划项目(21JK0578);陕西能源职业技术学院校级自然科学重点项目(2021KY01)。

摘　　要：目的探讨白虎加人参汤加减方对2型糖尿病(T2DM)大鼠骨骼肌葡萄糖转运蛋白4(GLUT4)表达及抗氧化应激作用的影响及机制。方法将SD大鼠随机分为对照组、模型组、阳性药组(二甲双胍,200 mg·kg^(-1))及白虎加人参汤加减方高、低剂量组(37.8、18.9 g·kg^(-1)),每组8只。采用高脂高糖饲料喂养4周后,一次性腹腔注射STZ 40 mg·kg^(-1)复制T2DM大鼠模型;灌胃给药(10 mL·kg^(-1)),每日1次,连续8周,T2DM大鼠继续给予高脂高糖饲料喂养。给药期间每2周测定1次空腹血糖(FBG)值;给药结束后,测定大鼠糖化血红蛋白(HbA1c)、血清胰岛素(INS)水平,计算胰岛素抵抗指数(HOMA-IR);测定大鼠胰腺组织中谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及丙二醛(MDA)水平;qRT-PCR法检测肌肉组织中GLUT4 mRNA表达水平;Western Blot法检测肌肉组织中GLUT4蛋白及胰腺组织中蛋白激酶B(AKT)/糖原合成激酶3β(GSK-3β)/核因子NF-E2相关因子(Nrf2)通路相关蛋白表达水平;免疫组织化学法检测胰腺组织中血红素加氧酶1(HO-1)、n-Nrf2蛋白表达水平。结果与对照组比较,模型组大鼠的FBG、HbA1c及HOMA-IR明显升高(P<0.05),血清INS水平明显降低(P<0.05);胰腺组织的CAT、SOD、GSH-Px水平均明显降低(P<0.05),MDA水平明显升高(P<0.05);肌肉组织中GLUT4蛋白及mRNA表达水平明显降低(P<0.05);胰腺组织中的p-AKT/AKT、p-GSK-3β/GSK-3β蛋白表达水平明显降低(P<0.05),n-Nrf2、n-Fyn、HO-1蛋白表达水平明显升高(P<0.05)。与模型组比较,白虎加人参汤加减方组大鼠第4、6、8周的FBG水平及HbA1c、HOMA-IR均明显降低(P<0.05),血清INS水平明显升高(P<0.05);胰腺组织中CAT、SOD、GSH-Px水平明显升高(P<0.05),MDA水平明显降低(P<0.05);肌肉组织中GLUT4蛋白及mRNA表达水平均明显升高(P<0.05);胰腺组织中p-AKT/AKT、p-GSK-3β/GSK-3β、n-Nrf2、HO-1蛋白表达水平明显升高(P<0.05),n-Fyn蛋白表达水平明�Objective To investigate the effects and mechanisms of Modified Baihu Plus Renshen Decoction on glucose transporter 4(GLUT4)expression in skeletal muscle and anti-oxidative stress in type 2 diabetes mellitus(T2DM)rats.Methods SD rats were randomly divided into control group,model group,positive drug group(Metformin,200 mg·kg^(-1))and high-and low-dose groups(37.8 and 18.9 g·kg^(-1))of Modified Baihu Plus Renshen Decoction,8 rats in each group.After 4 weeks of high-fat and high-sugar diet,the T2DM rat model was replicated by a one-time intraperitoneal injection of STZ 40 mg-kg';the drug was administered by gavage(10 mL·kg^(-1))once a day for 8 weeks,and the T2DM rats continued to be fed with high-fat and high-sugar diet.Fasting blood glucose(FBG)was measured every 2 weeks during the administration period;at the end of the administration period,the levels of glycated haemoglobin(HbAlc)and serum insulin(INS)were measured,and the index of insulin resistance(HOMA-IR)was calculated;and the levels of glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),catalase(CAT),and malondialdehyde(MDA)were determined in the pancreas tissue of the rats.The mRNA expression level of GLUT4 in muscle tissue were detected by qRT-PCR.The protein expression levels of GLUT4 in muscle tissue and protein kinase B(AKT)/glycogen synthase kinase 3β(GSK-3β)/nuclear factor NF-E2-related factor(Nrf2)pathway in pancreas were detected by Western Blot;the protein expression levels of heme oxygenase 1(HO-1)and n-Nrf2 in pancreatic tissue were detected by immunohistochemistry.Results Compared with the control group,the FBG,HbAlc and HOMA-IR of rats in the model group were significantly increased(P<0.05),and the serum INS level was significantly decreased(P<0.05);the CAT,SOD and GSH-Px levels of pancreatic tssue were allsignificantly decreased(P<0.05),and the MDA level was significantly increased(P<0.05);the protein and mRNA expression levels of CLUT4 were significantly reduced(P<0.05);protein expression levels of p-AKT/AKT,p-GSK-3β/GSK-3βin p

关 键 词：白虎加人参汤加减方 2型糖尿病 胰岛素抵抗 葡萄糖转运蛋白4 抗氧化应激 AKT/GSK-3β/Nrf2通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]