基于PKA/CREB/GLP-1信号通路探讨加味葛根芩连汤对糖尿病db/db小鼠胰腺损伤的影响  

Effects of Modified Gegen Qinlian Decoction on Pancreatic Injury of Diabetes Mellitus db/db Mice Based on PKA/CREB/GLP-1 Signaling Pathway

作　　者：柳荣 王佳慧 杨霞 高艳奎 刘苗 梁永林 朱向东 LIU Rong;WANG Jiahui;YANG Xia;GAO Yankui;LIU Miao;LIANG Yonglin;ZHU Xiangdong(School of Basic Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;School of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750000,China)

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

出　　处：《中国中医药信息杂志》2023年第9期74-81,共8页Chinese Journal of Information on Traditional Chinese Medicine

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(2022CMG02034);甘肃省优秀研究生“创新之星”项目(2022CXZX-734)。

摘　　要：目的观察加味葛根芩连汤对2型糖尿病db/db小鼠胰腺损伤的影响,基于PKA/CREB/GLP-1信号通路探讨其治疗糖尿病胰腺损伤的作用机制。方法将50只db/db小鼠随机分为模型组、二甲双胍组和加味葛根芩连汤高、中、低剂量组,每组10只。另取10只m/m小鼠作为空白组。加味葛根芩连汤高、中、低剂量组分别予加味葛根芩连汤31.9、19.1、6.4 g/kg灌胃,二甲双胍组予二甲双胍0.2 g/kg灌胃,空白组和模型组予蒸馏水灌胃,连续12周。给药结束后检测小鼠空腹血糖(FBG),进行口服葡萄糖耐量试验,检测小鼠糖化血红蛋白(HbA1c)含量,TUNEL法检测胰腺组织细胞凋亡情况,RT-qPCR检测胰腺组织G蛋白偶联胆汁酸受体5(TGR5)、蛋白激酶A(PKA)、环磷腺苷效应元件结合蛋白(CREB)、前蛋白转化酶1/3(PC1/3)、胰高血糖素样肽-1(GLP-1)mRNA表达,免疫组化法检测胰腺组织TGR5、PKA、p-PKA、CREB、p-CREB、PC1/3、GLP-1蛋白表达,免疫荧光染色检测胰腺组织TGR5/GLP-1、神经源性分化因子1(NeuroD1)/GLP-1共表达。结果与空白组比较,模型组小鼠FBG、HbA1c含量显著升高(P<0.01),口服葡萄糖耐量显著降低;胰腺组织细胞凋亡率显著升高(P<0.01),TGR5、PKA、CREB、PC1/3、GLP-1 mRNA表达显著降低(P<0.01),TGR5、PKA、p-PKA、CREB、p-CREB、PC1/3、GLP-1蛋白表达显著降低(P<0.01),TGR5/GLP-1、NeuroD1/GLP-1共表达显著降低(P<0.01)。与模型组比较,二甲双胍组和加味葛根芩连汤高、中剂量组小鼠FBG、HbA1c含量显著降低(P<0.01,P<0.05),口服葡萄糖耐量升高(P<0.01);各给药组小鼠胰腺组织细胞凋亡率显著降低(P<0.01),二甲双胍组和加味葛根芩连汤高、中剂量组小鼠胰腺组织TGR5、PKA、CREB、PC1/3、GLP-1 mRNA表达显著升高(P<0.01),TGR5/GLP-1、NeuroD1/GLP-1共表达显著升高(P<0.01),二甲双胍组和加味葛根芩连汤高剂量组胰腺组织TGR5、PKA、p-PKA、CREB、p-CREB、PC1/3、GLP-1蛋白表达显著升高(P<0.01)。结论�Objective To observe the effects of modified Gegen Qinlian Decoction on the pancreatic injury of type 2 diabetes mellitus;To explore its mechanism in the treatment of pancreatic injury in diabetes mellitus based on PKA/CREB/GLP-1 signaling pathway.Methods Totally 50 db/db mice were randomly divided into the model group,metformin group,modified Gegen Qinlian Decoction high-,medium-,and low-dosage groups,with 10 mice in each group.Another 10 m/m mice were selected as the blank group.The modified Gegen Qinlian Decoction high-,medium-,and low-dosage groups were given modified Gegen Qinlian Decoction 31.9,19.1,6.4 g/kg by gavage respectively,the metformin group was given metformin 0.2 g/kg by gavage,and the blank group and model group were given equal volume of distilled water by gavage.Each group was given continuous gavage for 12 weeks.The fasting blood glucose(FBG)was measured,an oral glucose tolerance test was performed,the contents of HbA1c were detected,apoptosis in pancreatic tissue was measured by TUNEL staining,TGR5,PKA,CREB,PC1/3,GLP-1 mRNA expression in pancreatic tissue were detected by RT-qPCR,the protein expression of TGR5,PKA,p-PKA,CREB,p-CREB,PC1/3 and GLP-1 in pancreatic tissue were detected by immunohistochemical method,coexpression of TGR5/GLP-1,NeuroD1/GLP-1 in pancreatic tissue were detected by immunofluorescence staining.Results Compared with the blank group,the contents of FBG and HbA1c in the model group significantly increased(P<0.01),while the oral glucose tolerance significantly decreased;cells apoptosis rate of pancreatic tissue significantly increased(P<0.01);TGR5,PKA,CREB,PC1/3,GLP-1 mRNA expressions were significantly decreased(P<0.01),TGR5,PKA,p-PKA,CREB,p-CREB,PC1/3,GLP-1 protein expressions significantly decreased(P<0.01),and TGR5/GLP-1,NeuroD1/GLP-1 co-expression was significantly decreased(P<0.01).Compared with the model group,the contents of FBG and HbA1c in the metformin group and modified Gegen Qinlian Decoction high-,and medium-dosage groups significantly decreased(P<0.01,P<0.05

关 键 词：2型糖尿病 加味葛根芩连汤 胰腺损伤 PKA/CREB/GLP-1信号通路 小鼠 

分 类 号：R285.5[医药卫生—中药学]