小檗碱调控AMPK-TSC1/LC3-Ⅱ信号通路促进高糖高脂应激下NIT-1胰岛β细胞的自噬  

Berberine induces autophagy in NIT-1 cells exposed by high glucose and palmitic acid through targeting the AMPK-TSC1/LC3-Ⅱ signaling pathway

作　　者：胡亚耘 陆付耳[1] 胡倩 易屏 HU Ya-yun;LU Fu-er;HU Qian;YI Ping(Institute of Integrated Traditional Chinese and Western Medicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Hubei Wuhan 430030,China;The First Hospital of Wuhan,Hubei Wuhan 430030,China;Department of Integrated Traditional Chinese and WesternMedicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Hubei Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院中西医结合研究所,湖北武汉430030 [2]武汉市中西医结合医院内分泌科,湖北武汉430030 [3]华中科技大学同济医学院附属同济医院中西医结合科,湖北武汉430030

出　　处：《中国医院药学杂志》2020年第1期29-37,共9页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金项目(编号:81673757、81573787)

摘　　要：目的:观察小檗碱(berberine, BBR)对高糖高脂应激条件下的NIT-1胰岛β细胞自噬的影响并探讨其潜在的分子机制。方法:分别采用高浓度葡萄糖(25 mmol·L^-1)和软脂酸(0.25 mmol·L^-1)诱导NIT-1胰岛β细胞建立高糖、高脂模型,予以小檗碱处理,用放射免疫法检测胰岛素水平,通过电镜观察自噬体的形成;然后予以腺苷酸活化蛋白激酶(AMPK)激动剂AICAR,腺苷酸活化蛋白激酶(AMPK)抑制剂Compound C,自噬诱导剂雷帕霉素(Rapa)进行干预,Western blot方法检测各组细胞AMPK的活化、自噬相关蛋白结节性硬化复合物1(TSC1)、微管相关蛋白1轻链蛋白3(LC3-Ⅱ)的表达。结果:在高糖高脂诱导的NIT-1胰岛β细胞中,小檗碱可以促进胰岛素的分泌和自噬体的形成,促进自噬相关蛋白TSC1,LC3-Ⅱ的表达,并活化其上游的AMPK。结论:小檗碱在高糖高脂的应激条件下可能通过促进自噬来发挥保护胰岛β细胞功能的作用,其分子机制可能与小檗碱调控AMPK-TSC1/LC3-II信号通路相关蛋白有关。OBJECTIVE To observe the effect of berberine(BBR) on autophagy in NIT-1 pancreatic β cells under high glucose and high fat stress and to explore its potential molecular mechanism.METHODS High glucose(25 mmol · L^-1) and palmitic acid(0.25 mmol · L^-1) were used to induce NIT-1 pancreatic β cells to establish high glucose and high fat models, respectively. Berberine was used to treat NIT-1 pancreatic β cells. Insulin levels were measured by radioimmunoassay, and autophagosome formation was observed by electron microscopy. Then, Adenosine 5‘-monophosphate(AMP)-activated protein kinase agonist AICAR, AMP-activated protein kinase(AMPK) inhibitor Compound C, and autophagy inducer rapamycin(Rapa) were used to intervene. Western blot was used to detect the activation of AMPK, the expression of autophagy-related protein nodular sclerosis complex 1(TSC1) and microtubule-associated protein 1 light chain protein 3(LC3-Ⅱ).RESULTS Berberine could promote insulin secretion and autophagosome formation, enhance the expression of autophagy related proteins TSC1, LC3-II and activate AMPK in NIT-1 pancreatic β cells induced by high glucose and high fat.CONCLUSION Berberine may play a role in protecting pancreatic β cells by promoting autophagy under high glucose and high fat stress conditions, and its molecular mechanism may be related to the regulation of AMPK-TSC1/LC3-Ⅱ signaling pathway related proteins by berberine.

关 键 词：小檗碱 自噬 NIT-1细胞 AMPK TSC1 LC3-Ⅱ 

分 类 号：R285[医药卫生—中药学]