加味葛根芩连汤对2型糖尿病db/db小鼠FXR/SHP/PPARα信号通路的影响  

Effect of Modified Gegen Qinliantang on FXR/SHP/PPARαSignaling Pathway in Type 2 Diabetic db/db Mice

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 Traditional Chinese Medicine(TCM),Jiangxi University of Chinese Medicine,Nanchang 330004,China;School of Basic Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;School of TCM,Ningxia Medical University,Yinchuan 750000,China)

机构地区：[1]江西中医药大学中医学院,南昌330004 [2]甘肃中医药大学基础医学院,兰州730000 [3]宁夏医科大学中医学院,银川750000

出　　处：《中国实验方剂学杂志》2023年第20期1-8,共8页Chinese Journal of Experimental Traditional Medical Formulae

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)。

摘　　要：目的:该研究拟通过观察加味葛根芩连汤对2型糖尿病(T2DM)模型db/db小鼠肝组织核受体法尼醇X受体/小异源二聚体伴侣/过氧化物酶体增殖物激活受体α(FXR/SHP/PPARα)信号通路相关蛋白表达水平的影响,探讨加味葛根芩连汤可能的作用机制。方法:将30只db/db小鼠随机分为模型组、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)),每组6只,另取6只m/m小鼠作为空白组,分别给予相应药物灌胃12周。检测小鼠体质量、空腹血糖(FBG)、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)含量,油红O染色观察肝脏脂质蓄积,过碘酸-雪夫(PAS)染色观察肝糖原沉积,硫酸铁铵染色观察回肠组织胆固醇沉积,蛋白免疫印迹法(Western blot)检测肝组织FXR、胆固醇7α-羟化酶(CYP7A1)、SHP、PPARα蛋白表达,酶联免疫吸附测定法(ELISA)检测血清游离脂肪酸(FFA)水平。结果:治疗结束后,与空白组比较,模型组小鼠体质量、FBG、FFA、TC、TG、LDL-C水平均显著升高(P<0.01),肝组织内有大量脂滴,肝糖原显著减少,回肠组织内有明显胆固醇堆积,肝组织FXR、SHP、PPARα蛋白表达显著降低,CYP7A1蛋白表达显著增高(P<0.01);与模型组比较,二甲双胍组及加味葛根芩连汤高、中剂量组小鼠体质量、FBG、FFA、TC、TG、LDL-C水平明显降低(P<0.05,P<0.01),HDL-C水平明显增加(P<0.05,P<0.01),肝细胞脂质蓄积减少,肝糖原显著增加,回肠组织胆固醇减少,肝组织FXR、SHP、PPARα蛋白表达显著增高,CYP7A1蛋白表达显著降低(P<0.01)。结论:加味葛根芩连汤可能通过调节FXR/SHP/PPARα信号通路抑制FFA水平,改善T2DM小鼠脂质代谢。Objective:To observe the effect of modified Gegen Qinliantang on the expression levels of proteins related to the farnesoid X receptor/small heterodimer partner/peroxisome proliferator-activated receptorα(FXR/SHP/PPARα)signaling pathway in the liver tissue of db/db model mice with type 2 diabetes mellitus(T2DM)and explore the underlying mechanism of action of modified Gegen Qinliantang.Method:Thirty db/db mice were randomly divided into model group,metformin group(0.2 g·kg^(-1)),and high-,medium-,and low-dose modified Gegen Qinliantang groups(31.9,19.1,6.4 g·kg^(-1)),with 6 mice in each group.An additional six m/m mice were assigned to the blank group.Respective drugs were administered via oral gavage for 12 weeks.Mouse body weight,fasting blood glucose(FBG),total cholesterol(TC),triglyceride(TG),highdensity lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)levels were measured.Oil red O staining was used to observe hepatic lipid accumulation and periodic acid-schiff(PAS)staining was used to assess hepatic glycogen deposition.Ammonium ferric sulfate staining was used to observe cholesterol deposition in intestinal tissues.Western blot was employed to detect the expression of FXR,cholesterol 7α-hydroxylase(CYP7A1),SHP,and PPARαproteins in liver tissues,and enzyme-linked immunosorbent assay(ELISA)was used to measure serum free fatty acid(FFA)levels.Result:At the end of the treatment,compared with the blank group,the model group exhibited significant increases in mouse body weight,FBG,FFA,TC,TG,and LDL-C levels(P<0.01),along with significant hepatic lipid droplets,reduced hepatic glycogen,noticeable cholesterol accumulation in intestinal tissues,significantly decreased expression of FXR,SHP,PPARαproteins,and significantly increased expression of CYP7A1 protein in liver tissues(P<0.01).Compared with the model group,the metformin group and the high-and medium-dose modified Gegen Qinliantang groups demonstrated significant reductions in mouse body weight,FBG,FFA,TC,TG,LDL-C levels(P<0

关 键 词：2型糖尿病 加味葛根芩连汤 法尼醇X受体/小异源二聚体伴侣/过氧化物酶体增殖物激活受体α 

分 类 号：R2-0[医药卫生—中医学] R33R587.1R289