葛根素通过Fetuin B-AMPK/ACC信号通路减轻2型糖尿病小鼠肝脏胰岛素抵抗  

Puerarin alleviates insulin resistance in type 2 diabetic mice by modulating fetuin B-AMPK/ACC signaling pathway in the liver

作　　者：高俊凤 刘曼曼[2] 郭召平[2] 胡春平 冯珍凤 严军 GAO Junfeng;LIU Manman;GUO Zhaoping;HU Chunping;FENG Zhenfeng;YAN Jun(Graduate School,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Cooperation Research Center of Shanghai University of Traditional Chinese Medicine//Department of Endocrinology,Traditional Chinese Medicine Hospital of Jiading District,Shanghai 201899,China)

机构地区：[1]上海中医药大学研究生院,上海201203 [2]上海中医药大学联合培养单位//上海市嘉定区中医医院内分泌科,上海201899

出　　处：《南方医科大学学报》2021年第6期839-846,共8页Journal of Southern Medical University

基　　金：上海市嘉定区科委项目(JDKW-2019-W17);上海市五大新城中医药特色专科支持计划(2020-2023);上海市进一步加快中医药事业发展三年行动计划[ZY(2018-2020)-FWTX-4011]。

摘　　要：目的探讨葛根素通过Fetuin B-AMPK/ACC信号通路对高脂饮食联合链脲佐菌素诱导的2型糖尿病小鼠肝脏胰岛素抵抗的调控机制。方法 C57BL/6J小鼠随机选取10只正常饮食作为正常对照组,其余40只通过高脂饲料喂养联合腹腔注射100 mg/kg链脲佐菌素建立2型糖尿病小鼠模型,造模成功后采用随机数字表法将小鼠分为模型对照组、葛根素低剂量组(Pue-50 mg/kg)、葛根素中剂量组(Pue-100 mg/kg)和葛根素高剂量组(Pue-200 mg/kg),干预8周。末次给药后,尾静脉取血检测各组小鼠空腹血糖(FBG),取血和肝组织;ELISA法检测各组小鼠血清空腹胰岛素(FINS),肝脏甘油三酯、胆固醇、游离脂肪酸水平;HE染色法观察小鼠肝脏病理变化;免疫组化法检测小鼠肝脏Fetuin B含量;实时荧光定量聚合酶链式反应法检测小鼠肝脏Fetuin B、AMPK、ACC mRNA表达;Western blot法检测小鼠肝脏Fetuin B、AMPKα1、ACC、P-AMPKαT183/T172、P-ACCS79蛋白表达。结果末次给药后,Pue-100 mg/kg、Pue-200 mg/kg剂量组甘油三酯、胆固醇、游离脂肪酸及FBG低于模型对照组(P<0.01);葛根素不同剂量给药组较正常对照组,FINS和胰岛素抵抗指数(HOMA-IR)降低(P<0.01)。与正常对照组比较,模型对照组小鼠肝脏Fetuin B、ACC mRNA表达明显上升,AMPK mRNA表达降低(P<0.01);Fetuin B、ACC蛋白表达升高,AMPKα1、PAMPKαT183/T172、P-ACC S79蛋白表达降低(P<0.01);病理学观察到肝脏脂肪变性明显,肝脏Fetuin B表达含量增加。与模型对照组比较,葛根素呈剂量依赖性抑制肝脏Fetuin B、ACC mRNA表达和Fetuin B、ACC蛋白表达,上调AMPK mRNA表达和AMPKα1、P-AMPKαT183/T172、P-ACC S79蛋白表达(P<0.01)。Pue-50 mg/kg给药组上调AMPK mRNA表达,下调ACC mRNA表达差异无统计学意义(P>0.05)。结论葛根素可能通过调节肝脏Fetuin B-AMPK/ACC信号通路减轻2型糖尿病小鼠胰岛素抵抗,进一步改善糖脂代谢。Objective To explore the role of fetuin B-AMPK/ACC signaling pathway in mediating the effect of puerarin on hepatic insulin resistance in mice with type 2 diabetes mellitus(T2DM).Methods Forty C57BL/6J mouse models of T2DM induced by high-fat diet and intraperitoneal injection of streptozotocin were randomized into diabetic model(HFD)group and 3 puerarin groups for treatment with low-,moderate-and high-dose puerarin(50,100 and 200 mg/kg,respectively),with another 10 mice fed a normal diet as the control group.After treatment for 8 weeks,the mice were examined for fasting blood glucose(FBG),fasting insulin(FINS),liver triglycerides(TG),cholesterol(TC)and free fatty acids(FFA)levels.The expression of fetuin B in the liver was detected by immunohistochemistry.RT-qPCR was used to detect the expressions of fetuin B,AMPK,and ACC mRNA in the liver,and the protein expressions of fetuin B,AMPKα1,ACC,P-AMPKαT183/T172,and P-ACC S79 were determined with Western blotting.Results Treatment with moderate-and high-dose puerarin significantly lowered TG,TC,FFA and FBG levels in diabetic mice(P<0.01).Puerarin at all the 3 doses significantly lowered FINS and HOMA-IR of the mice(P<0.01).In diabetic mice,hepatic expressions of fetuin B and ACC mRNA increased and AMPK mRNA decreased significantly(P<0.01);the protein expressions of fetuin B and ACC increased while those of AMPKα1,P-AMPKαT183/T172 and P-ACC S79 decreased significantly(P<0.01).Puerarin dose-dependently inhibited the mRNA and protein expressions of fetuin B and ACC,increased AMPK mRNA and protein expressions of AMPKα1,P-AMPKαT183/T172,and P-ACC S79,and lowered fetuin B content in the liver of diabetic mice(P<0.01).Conclusion Puerarin alleviates insulin resistance and improves glucolipid metabolism in T2DM mice by modulating hepatic fetuin B-AMPK/ACC signaling pathway.

关 键 词：葛根素 Fetuin B AMPK/ACC 胰岛素抵抗 糖脂代谢 

分 类 号：R285.5[医药卫生—中药学]