文献类型：期刊文献

中文题名：葛根芩连汤对db/db糖尿病小鼠SIRT1/FoxO1自噬通路的影响

英文题名：Effect of Gegenqinlian Decoction on SIRT1/FoxO1 Autophagy Pathway in db/db Diabetic Mice

作者：张媛媛[1];朱向东[2];樊俐慧[1];苏菲[1];关晓文[1];翟艳会[1];曹力仁[1];周楠[1];霍敏峰[1];梁建庆[1]

第一作者：张媛媛

机构：[1]甘肃中医药大学,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：14

起止页码：30

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);甘肃省教育科技创新项目(2021CYZC-03)。

语种：中文

中文关键词：葛根芩连汤;2型糖尿病;胰岛素抵抗;沉默信息调节因子1(SIRT1)/叉头框蛋白O1(FoxO1)信号通路;自噬

外文关键词：Gegenqinlian decoction;Type 2 diabetes;Insulin resistance;SIRT1/FoxO1 signaling pathway;Autophagy

摘要：目的:探究葛根芩连汤是否通过调控沉默信息调节因子1(SIRT1)/叉头框蛋白O1(FoxO1)自噬通路减轻db/db糖尿病小鼠胰岛素抵抗。方法:选取SPF级自发性2型糖尿病动物模型db/db小鼠75只及对照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8周,1次/d,使用罗氏血糖仪检测空腹血糖(FBG),全自动生化分析仪检测血清中高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)和总胆固醇(TC)水平,酶联免疫吸附测定法(ELISA)检测血清空腹胰岛素(INS)水平,计算胰岛素抵抗指数(HOMA-IR);采用蛋白免疫印迹法(Western blot)检测肝脏组织中Beclin-1、微管相关蛋白1轻链3(LC3)及SIRT1/FoxO1自噬通路相关蛋白的表达;免疫组化检测肝脏组织中SIRT1、FoxO1、Beclin-1、LC3B蛋白的表达;透射电镜观察肝脏自噬小体形成。结果:与正常组比较,模型组FBG、FINS、HOMA-IR、TC、TG、LDL-C、HDL-C水平显著升高(P<0.01),肝脏组织SIRT1、Beclin-1、LC3蛋白表达显著升高(P<0.01),FoxO1显著升高(P<0.01),透射电镜显示模型组自噬小体最多;与模型组比较,二甲双胍组、葛根芩连汤低、中、高剂量组血清FBG、FINS、HOMA-IR、TC、TG、LDL-C、HDL-C水平明显降低(P<0.05,P<0.01),肝脏组织SIRT1、Beclin-1、LC3蛋白表达明显降低(P<0.05,P<0.01),FoxO1显著降低(P<0.01),透射电镜显示用药组自噬程度有所减轻;与二甲双胍组比较,葛根芩连汤中、高剂量组FBG、FINS、TG水平显著降低(P<0.01),肝脏组织SIRT1、Beclin-1、LC3蛋白表达明显降低(P<0.05,P<0.01),FoxO1显著降低(P<0.01),葛根芩连汤高剂量HOMA-IR、TC、LDL-C、HDL-C水平明显降低(P<0.05,P<0.01),透射电镜显示葛根芩连汤中、高剂量组自噬小体明显减少。结论:葛根芩连汤可显著改善糖脂代谢紊乱,通过激活SIRT1/FoxO1自噬通路减轻db/db小鼠胰岛素抵抗从而防治2型糖尿病。
Objective:To validate the alleviating effect of Gegen Qinliantang(GGQLT)on insulin esistance in db/db diabetic mice by regulating the silent information regulator 1(SIRT1)/forkhead transcription factor O1(FoxO1)autophagy pathway.Method:Seventy-five SPF-grade spontaneous type 2 diabetic db/db mice and 15 control db/m mice were selected and maintained on regular feed for one week before measuring blood glucose.They were randomly divided into six groups,with 15 mice in each group.The groups included a normal group(physiological saline,0.2 g·kg^(-1)),a metformin group(0.2 g·kg^(-1)),high-,medium-,and lowdose GGQLT groups(31.9,19.1,6.9 g·kg^(-1)),and a model group(physiological saline,0.2 g·kg^(-1)).They were orally treated with corresponding drugs for eight weeks,once daily.Fasting blood glucose(FBG)was measured using a Roche glucometer.Serum levels of high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),triglyceride(TG),and total cholesterol(TC)were measured using an automated biochemical analyzer.Fasting serum insulin(INS)levels were determined using enzyme-linked immunosorbent assay(ELISA),and the homeostasis model assessment of insulin resistance(HOMA-IR)was calculated.Western blot was used to detect the expression of Beclin-l,microtubule-associated protein 1 light chain 3(LC3),and SIRT1/FoxO1 autophagy pathway-related proteins in liver tissues.Immunohistochemistry was performed to assess the expression of SIRT1,FoxO1,Beclin-1,and LC3B proteins in liver tissues.Transmission electron microscopy was used to observe the formation of autophagosomes in the liver.Result:Compared with the normal group,the model group showed significant increases in FBG,FINS,HOMA-IR,TC,TG,LDL-C,and HDL-C levels(P<0.01),and significant increases in the expression of SIRT1,Beclin-1,LC3,and FoxO1 proteins in liver tissues(P<0.01).Transmission electron microscopy revealed the highest number of autophagosomes in the model group.Compared with the model group,the metformin group and the low-,medium-,and high-dose GGQLT groups showed significant decreases in serum FBG,FINS,HOMA-IR,TC,TG,LDL-C,and HDL-C levels(P<0.05,P<0.01),significant decreases in the expression of SIRT1,Beclin-1,LC3(P<0.05,P<0.01),and up-regulated FoxO1 protein(P<0.01).Transmission electron microscopy showed a reduction in the degree of autophagy in the treatment groups.Compared with the metformin group,the medium-and high-dose GGQLT groups showed significant decreases in FBG,FINS,and TG levels(P<0.01),significant decreases in the expression of SIRT1,Beclin-1,and LC3 in liver tissues(P<0.05,P<0.01),and reduced FoxO1 protein(P<0.01).The high-dose GGQLT group showed reduced HOMAIR,TC,LDL-C,and HDL-C levels(P<0.05,P<0.01).Transmission electron microscopy revealed a significant reduction in autophagosomes in the medium-and high-dose GGQLT groups.Conclusion:GGQLT can significantly improve glucose and lipid metabolism disorders,alleviate insulin resistance in db/db mice,and prevent and treat type 2 diabetes by activating the SIRT1/FoxO1 autophagy pathway.