文献类型：期刊文献

中文题名：基于AMPK-FoxO3a自噬轴探讨葛根芩连汤改善2型糖尿病db/db小鼠肝脏脂质异位蓄积的机制

英文题名：Mechanism of Gegen Qinliantang in Improving Ectopic Lipid Accumulation in Liver of db/db Mice with Type 2 Diabetes Mellitus by Regulating AMPK-FoxO3a Autophagy Axis

作者：梁建庆[1];张媛媛[1];朱向东[2];司晓丽[1]

第一作者：梁建庆

机构：[1]甘肃中医药大学基础医学院,甘肃省中医方药挖掘与创新转化重点实验室,糖尿病实验室,兰州730000;[2]宁夏医科大学,银川750004

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）|甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2023

卷号：29

期号：18

起止页码：1

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：甘肃省自然科学基金一般项目(2021-0405-JCC-0791);宁夏回族自治区重点研发计划项目(2022CMG02034)。

语种：中文

中文关键词：葛根芩连汤;2型糖尿病;腺苷酸活化蛋白激酶(AMPK)-叉头框蛋白O3a(FoxO3a)自噬轴;脂质的异位蓄积

外文关键词：Gegen Qinliantang;type 2 diabetes mellitus;adenosine monophosphate-activated protein kinase(AMPK)-forkhead box O3a(FoxO3a)autophagy axis;ectopic lipid accumulation

摘要：目的:探讨葛根芩连汤调控腺苷酸活化蛋白激酶(AMPK)-叉头框蛋白O3a(FoxO3a)自噬轴改善2型糖尿病db/db小鼠肝脏脂质异位蓄积的机制研究,为阐明葛根芩连汤的降糖机制及推广应用提供科学依据。方法:选取SPF级自发性2型糖尿病动物模型db/db小鼠75只及正常db/m小鼠15只予维持饲料喂养1周后检测血糖,随机分为6组,每组15只。正常组(生理盐水0.2 g·kg^(-1))、二甲双胍组(0.2 g·kg^(-1))、葛根芩连汤高、中、低剂量组(31.9、19.1、6.9 g·kg^(-1))及模型组(生理盐水0.2 g·kg^(-1)),连续灌胃给药12周,1次/d,检测各组小鼠空腹血糖(FBG)、糖化血红蛋白(HbA1c),酶联免疫吸附测定法(ELISA)检测血清空腹胰岛素(FINS)、游离脂肪酸(FFA)水平,苏木素-伊红(HE)染色观察肝组织病理改变,使用蛋白免疫印迹法(Western blot)检测肝脏组织中磷酸化(p)-AMPK、p-FoxO3a及自噬相关蛋白微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)、p62蛋白的表达;免疫荧光检测肝脏组织中缺氧诱导因子-1α(HIF-1α)蛋白表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测肝组织AMPK、FoxO3a、LC3 mRNA表达。结果:与正常组比较,模型组小鼠肝脏组织有病理学改变;模型组小鼠FBG、HbA1c、FINS和FFA水平显著升高(P<0.01);肝脏组织中p-AMPK、p62、HIF-1α蛋白表达水平显著升高,p-FoxO3a、LC3Ⅱ的蛋白表达显著降低(P<0.01);模型组小鼠肝脏组织AMPK mRNA表达显著降低,FoxO3a表达显著升高(P<0.01)。与模型组比较,各给药组肝脏组织病变程度减轻;各给药组FBG、HbA1c、FINS和FFA水平有所降低(P<0.01);葛根芩连汤中、高剂量组p-AMPK、p62、HIF-1α蛋白表达有所增加,p-FoxO3a的表达呈剂量依赖性降低,二甲双胍组、葛根芩连汤高剂量组LC3Ⅱ的表达有所升高(P<0.01);给药组AMPK mRNA的表达呈剂量依赖性增加,FoxO3a的表达呈剂量依赖性降低(P<0.01)。结论:葛根芩连汤可改善2型糖尿病db/db小鼠肝脏脂质异位蓄积,可能与调控AMPK-FoxO3a自噬轴相关。
Objective:To explore the mechanism of Gegen Qinliantang(GQT)in improving ectopic lipid accumulation in the liver of db/db mice with type 2 diabetes mellitus(T2DM)by regulating the adenosine monophosphate-activated protein kinase(AMPK)-forkhead box O3a(FoxO3a)autophagy axis,to provide a scientific basis for clarifying the hypoglycemic mechanism of GQT and its clinical application.Method:Seventy-five spontaneous T2DM db/db mice and 15 normal db/m mice were selected and maintained on a regular diet for one week,followed by the measurement of blood glucose.They were then randomly divided into six groups,with 15 mice in each group,including normal group(0.2 g·kg^(-1)saline),metformin group(0.2 g·kg^(-1)),high-,medium,and low-dose GQT group(31.9,19.1,6.9 g·kg^(-1)),and model group(0.2 g·kg^(-1)saline).The mice were orally administered the corresponding drugs once daily for 12 weeks.Fasting blood glucose(FBG)and glycated hemoglobin(HbA1c)were detected.Fasting insulin(FINS)and free fatty acid(FFA)levels were measured by enzyme-linked immunosorbent assay(ELISA).Pathological changes in liver tissues were observed by hematoxylin-eosin(HE)staining.The protein expression levels of phosphorylated(p)-AMPK,p-FoxO3a,and autophagy-related proteins microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ)and p62 were detected using Western blot.Immunofluorescence was used to detect the expression of hypoxia-inducible factor-1α(HIF-1α)in liver tissues.Real-time polymerase chain reaction(Real-time PCR)was performed to detect the mRNA expression of AMPK,FoxO3a,and LC3 in liver tissues.Result:Compared with the normal group,the model group showed pathological changes in liver tissues,increased FBG,HbA1c,FINS,and FFA levels(P<0.01),increased protein expression levels of p-AMPK,p62,and HIF-1α,decreased protein expression levels of p-FoxO3a and LC3Ⅱin liver tissues(P<0.01),decreased mRNA expression of AMPK,and increased expression of FoxO3a(P<0.01).Compared with the model group,the treatment groups showed relieved liver tissue lesions and decreased FBG,HbA1c,FINS,and FFA levels(P<0.01).The expression of p-AMPK,p62,and HIF-1αincreased,while the expression of p-FoxO3a showed a dose-dependent decrease in the high-dose GQT group.The expression of LC3Ⅱincreased in the metformin group and the high-dose GQT group(P<0.01).The mRNA expression of AMPK showed a dose-dependent increase,and the expression of FoxO3a showed a dosedependent decrease in the treatment groups(P<0.01).Conclusion:GQT can improve ectopic lipid accumulation in the liver of T2DM db/db mice,which may be related to the regulation of the AMPK-FoxO3a autophagy axis.