二甲双胍对高糖状态下大鼠肾小球系膜细胞增殖的作用及机制  

Effects of metformin on the proliferation of rat mesangial cells induced by high glucose

作　　者：李凌云[1,2] 游娜[1] 徐家蓉[1] 缪珩[1] 鲁一兵[1] 叶小龙[1] 

机构地区：[1]南京医科大学第二附属医院内分泌科,江苏南京210011 [2]南京市江宁医院内分泌科,江苏南京211100

出　　处：《南京医科大学学报（自然科学版）》2011年第4期487-490,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京市医学科技发展基金(YKK08136);南京医科大学第二附属医院青年科技启动基金(QN200913)

摘　　要：目的:探讨二甲双胍对高糖状态下大鼠肾小球系膜细胞增殖的作用以及可能的作用机制。方法:体外培养大鼠肾小球系膜细胞(HBZY-1),①分为正常对照(NC)组(葡萄糖浓度5.6 mmol/L)、NC+二甲双胍组(葡萄糖浓度5.6 mmol/L+二甲双胍浓度200 mmol/L)、高糖(HG)组(葡萄糖浓度30 mmol/L)、HG+二甲双胍组(葡萄糖浓度30 mmol/L+二甲双胍,浓度分别是50、100、200、400 mmol/L),作用时间分别为6、12、24 h。MTT法检测各组HBZY-1细胞增殖的情况;②分为NC组、NC+二甲双胍组、HG组和HG+二甲双胍组(二甲双胍浓度200 mmol/L),作用时间12 h,用Western blot;法检测各组细胞内转化生长因子-β1(TGF-β1)、p-AMPK和AMPK蛋白表达的变化。结果:①与NC组相比,高糖刺激12 h后HBZY-1增殖明显,差异有显著性(P<0.01);与HG组相比,HG加二甲双胍干预12 h后,随着二甲双胍的浓度逐渐增加,对HBZY-1增殖的抑制程度逐渐加大,差异均有显著性(P均<0.01);NC+二甲双胍组与NC组相比,两组之间HBZY-1增殖差异没有显著性(P>0.05);②与NC组相比,高糖刺激12 h后HBZY-1细胞TGF-β1的表达增高而p-AMPK表达水平明显下降,差异均具有显著性(P均<0.01)。与HG组相比,加用二甲双胍干预后,TGF-β1的表达水平有明显下降,p-AMPK表达水平有显著升高,差异均具有显著性(P均<0.01)。4组之间AMPK表达差异无显著性(P>0.05)。结论:高糖环境下HBZY-1增殖明显,二甲双胍可能通过增加肾小球系膜细胞p-AMPK的表达、降低TGF-β1的表达抑制HBZY-1的增殖。Objective：To investigate the effect of metformin on the proliferation of rat mesangial cells induced by high glucose and the possible mechanism.Methods：The rat messangial cells（HBZY-1） were cultured in vitro.①Cultured HBZY-1 cells were divided into four groups：normal glucose（5.6 mmol/L）,normal glucose plus metformin（200 mmol/L）,high glucose（30 mmol/L）,high glucose plus metformin at 50,100,200,400 mmol/L,respectively,and cultured for 6,12,24 h.The proliferation of HBZY-1 cells was detected by micro-tetrazolium salt colorimetric enzyme reaction test（MTT）.②Cultured HBZY-1 cells were divided into four groups：normal glucose（5.6 mmol/L）,normal glucose plus metformin（200 mmol/L）,high glucose（30 mmol/L）,high glucose plus metformin（200 mmol/L）,and cultured for 12 h.The protein levels of TGF-β1,p-AMPK and AMPK were measured by Western blot.Results：As compared to normal cells,exposure of HBZY-1 cells to D-glucose at concentration of 30 nmol/L caused a time-dependent increase in the proliferation of HBZY-1 cells and reached the highest level at approximately 12 h.The addition of metformin significantly attenuated the high glucose-enhanced proliferation of HBZY-1 cells（P 0.01） in a dose-dependent manner.But no notable difference was found between normal glucose plus metformin group and normal glucose group（P 0.05）.As compared to normal cells,the levels of TGF-β1 protein significantly increased,and the levels of p-AMPK significantly decreased in high glucose group（P 0.01）.Metformin significantly attenuated the increase of TGF-β1 expression and increased the expression of p-AMPK in HBZY-1 cells exposured to high D-glucose（P 0.01）.Conclusion：High glucose contributes to an increased proliferation of HBZY-1 cells.Metformin can significantly reverse high glucose up-regulated proliferation of HBZY-1 cells through inhibiting the expression of TGF-β1 protein and increasing the expression of p-AMPK protein in HBZY-1 cells.

关 键 词：二甲双胍 大鼠肾小球系膜细胞 高糖 增殖 转化生长因子-β1 AMPK 

分 类 号：R587.1[医药卫生—内分泌]