文献类型：期刊文献

中文题名：加味葛根芩连汤对2型糖尿病db/db小鼠胰腺组织TGR5/cAMP/GLP-1信号通路的影响

英文题名：Effect of Modified Gegen Qinliantang on TGR5/c AMP/GLP-1 Signaling Pathway in Pancreatic Tissue of Type 2 Diabetes Mellitus db/db Mice

作者：柳荣[1];杨霞[1];高艳奎[1];王佳慧[1];梁永林[1];朱向东[2]

第一作者：柳荣

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]宁夏医科大学中医学院,银川750000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2023

卷号：29

期号：4

起止页码：25

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：甘肃省教育厅产业支撑计划项目(2021CYZC-03);宁夏回族自治区重点研发计划项目(2022CMG02034);甘肃省优秀研究生“创新之星”项目(2022CXZX-734)。

语种：中文

中文关键词：2型糖尿病;加味葛根芩连汤;胆汁酸;G蛋白偶联胆汁酸受体5(TGR5)/环磷腺苷(cAMP)/胰高糖素样肽-1(GLP-1)信号通路

外文关键词：type 2 diabetes mellitus;modified Gegen Qinliantang;bile acid;G protein-coupled receptor 5/cyclic adenosine monophosphate/glucagon-like peptide-1

摘要：目的:探讨加味葛根芩连汤对肥胖型2型糖尿病(T2DM)模型小鼠血糖血脂及胰腺组织中G蛋白偶联胆汁酸受体5(TGR5)相关途径的影响。方法:将10只7周龄无特定病原体(SPF)级雄性m/m小鼠及50只7周龄SPF级雄性db/db小鼠在SPF级实验室适应性喂养1周。m/m小鼠作为空白组。成模后随机分为5组,每组10只,分别作为模型组、二甲双胍组(0.2 g·kg^(-1))、加味葛根芩连汤高、中、低剂量组(31.9、19.1、6.4 g·kg^(-1)),灌胃体积均为10 mL·kg^(-1),模型组和空白组灌服等体积蒸馏水,1次/d,连续12周。定期检测小鼠空腹血糖(FBG)。药物干预12周后,检测血清中糖化血清蛋白(GSP)、血清葡萄糖(GLU)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)水平;采用苏木素-伊红(HE)染色观察各组小鼠胰腺组织病理改变,蛋白免疫印迹法(Western blot)检测胰腺组织TGR5、蛋白激酶A(PKA)、磷酸化(p)-PKA、环磷腺苷效应元件结合蛋白(CREB)、p-CREB、前蛋白转化酶1/3(PC1/3)、胰高糖素样肽-1(GLP-1)蛋白的表达水平,酶联免疫吸附测定法(ELISA)检测胰腺组织环磷腺苷(cAMP)的含量。结果:与空白组比较,模型组小鼠胰腺组织出现病理学改变;小鼠FBG、GSP、GLU、TC、TG、LDL-C水平显著增高(P<0.01),HDL-C水平明显降低(P<0.05);胰腺组织中TGR5、p-PKA(Thr197)/PKA、p-CREB(Ser133)/CREB、PC1/3、GLP-1蛋白表达水平显著降低(P<0.01);胰腺组织中cAMP的含量显著降低(P<0.01)。与模型组比较,治疗组胰腺组织病变程度减轻;加味葛根芩连汤高剂量组及二甲双胍组均能够明显降低db/db小鼠FBG、GSP、GLU、TC、TG、LDL-C水平(P<0.05,P<0.01),显著增高db/db小鼠HDL-C水平(P<0.01);除加味葛根芩连汤中剂量组GLP-1蛋白外,加味葛根芩连汤高、中剂量组及二甲双胍组TGR5、p-PKA(Thr197)/PKA、p-CREB(Ser133)/CREB、PC1/3、GLP-1蛋白表达水平均有不同程度的增加(P<0.05,P<0.01);加味葛根芩连汤高、中剂量组及二甲双胍组胰腺组织中cAMP的含量明显增高(P<0.05,P<0.01)。结论:加味葛根芩连汤能够改善T2DM模型db/db小鼠的葡萄糖稳态,其机制可能与调控TGR5/cAMP/GLP-1信号通路相关蛋白表达有关。
Objective:To discuss the effect of modified Gegen Qinliantang(MGQT)on blood glucose and lipids and Takeda G protein-coupled receptor 5(TGR5)-related pathways in pancreatic tissue of obese type 2diabetes mellitus(T2DM)mice.Method:A total of 10 male specific pathogen free(SPF)m/m mice(7 weeks old)and 50 male SPF(7 weeks old)were adaptively fed for one week in SPF laboratory.The m/m mice were included in the blank group.T2DM was induce d in the 50 db/db mice.The model mice were randomized into the model group,metformin group(0.2 g·kg^(-1)),high-dose,medium-dose,and low-dose(31.9,19.1,6.4 g·kg^(-1))MGQT groups,with 10 in each group,and the drug dose was10 mL·kg^(-1).The model group and the blank group received distilled water of the same volume.The administration lasted 12 weeks(once/day).Fasting blood glucose(FBG)was detected regularly.After 12 weeks of administration,serum levels of glycated serum protein(GSP),serum glucose(GLU),total cholesterol(TC),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)were detected.Pathological changes in the pancreatic tissue were based on hematoxylin-eosin(HE)staining.Western blot was used to determine the protein expression of TGR5,protein kinase A(PKA),phosphorylated(p)-PKA,cyclic-AMP response element binding protein(CREB),p-CREB,proprotein convertase 1/3(PC1/3),and glucagon-like peptide-1(GLP-1)in pancreatic tissues.The level of cyclic adenosine monophosphate(cAMP)in pancreatic tissue was determined by enzyme-linked immunosorbent assay(ELISA).Result:Compared with the blank group,the model group had pathological changes in pancreatic tissue,high levels of FBG,GSP,GLU,TC,TG,and LDL-C(P<0.01),low level of HDL-C(P<0.05),low protein expression of TGR5,p-PKA(Thr197)/PKA,p-CREB(Ser133)/CREB,PC1/3,and GLP-1 in pancreatic tissue(P<0.01),and low content of cAMP in the pancreas(P<0.01).Pancreatic tissue lesion in the treatment groups were milder than that in the model group.Both the high-dose MGQT and metformin can reduce the levels of FBG,GSP,GLU,TC,TG,and LDL-C in db/db mice(P<0.05,P<0.01)and increase the level of HDL-C(P<0.01).Except the GLP-1 protein in the medium-dose MGQT group,the protein expression of TGR5,p-PKA(Thr197)/PKA,p-CREB(Ser133)/CREB,PC1/3,and GLP-1in the high-dose and medium-dose MGQT groups and the metformin group increased compared with that in the model group(P<0.05,P<0.01).The content of cAMP in the pancreatic tissue of the high-dose and medium-dose MGQT groups and the metformin group was raised compared with that in model group(P<0.05,P<0.01).Conclusion:MGQT can improve the glucose homeostasis in db/db mice with T2DM by regulating TGR5/cAMP/GLP-1 signaling pathway-related protein expression.