南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (6): 839-846.doi: 10.12122/j.issn.1673-4254.2021.06.05

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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

高俊凤，刘曼曼，郭召平，胡春平，冯珍凤，严 军   

  • 出版日期:2021-06-20 发布日期:2021-07-02

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

  • Online:2021-06-20 Published:2021-07-02

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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、P-AMPKαT183/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-AMPK αT183/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ACCmRNA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。

关键词:

Abstract: 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.

Key words: puerarin; fetuin B; AMPK/ACC; insulin resistance; glucolipid metabolism