西格列汀对AGEs作用下系膜细胞自噬的影响  

Effect of sitagliptin on autopaghy in mesangial cells induced by AGEs

作　　者：江颖娟[1] 蒋作锋[1] 吴小兰 黄珮[1] 吴文法[1] 余慧文[1] JIANG Ying-juan;JIANG Zuo-feng;WU Xiao-lan;HUANG Pei;WU Weng-fa;Yu Hui-wen(Department of General Medicine, Guangzhou Red Cross Hospital,School of Medicine,Jinan University,Guangzhou 510220, China)

机构地区：[1]暨南大学医学院附属广州红十字会医院全科医学科,广东广州510220

出　　处：《中国病理生理杂志》2017年第8期1455-1459,共5页Chinese Journal of Pathophysiology

基　　金：广州市卫生和计划生育科技一般引导项目(No.20161A011020)

摘　　要：目的:探讨西格列汀对晚期糖基化终末产物(advanced glycation end products,AGEs)作用下系膜细胞细胞外基质和自噬相关蛋白表达的影响。方法:培养大鼠肾小球系膜细胞,共分5组,分别为正常对照(control)组、AGE组和不同浓度(5、10和20μmol/L)西格列汀组。培养48 h后采用MTT法测定细胞活力,采用ELISA法检测细胞培养上清胶原蛋白Ⅳ(collagen IV,Col IV)含量的变化。采用Western blot检测自噬相关蛋白beclin-1、腺苷酸活化蛋白激酶(adenosine monophosphate-activated protein kinase,AMPK)、p-AMPK、p70S6K和p-p70S6K的蛋白水平。结果:与control组比较,AGEs可明显引起系膜细胞活力和Col IV表达增加,不同浓度的西格列汀均可明显抑制AGEs诱导的系膜细胞活力和Col IV表达的增加;与control组比较,AGEs可引起系膜细胞自噬相关蛋白beclin-1表达和AMPK磷酸化水平下降,p70S6K磷酸化水平增加;不同浓度的西格列汀均可促进系膜细胞自噬相关蛋白beclin-1表达及AMPK磷酸化,抑制p70S6K磷酸化,并呈一定的浓度依赖性。结论:西格列汀可能通过引起系膜细胞的自噬发挥肾脏保护作用。AIM:To investigate the effect of sitagliptin on the autopaghy and the expression of extracellular matrix in mesangial cells induced by advanced glycation end products(AGEs).METHODS:The cells were divided into5groups:control group,AGE group,and sitagliptin(5,10and20μmol/L)groups.After48h,the cell viability was measured by MTT assay,and the content of collagen(Col)Ⅳin the supernatant of the cell culture was detected by ELISA.The protein levels of beclin-1,adenosine monophosphate-activated protein kinase(AMPK),p-AMPK,p70S6K and p-p70S6K were determined by Western blot.RESULTS:Compared with control group,the viability and the expression of Col IV induced by AGEs in the cultured mesangial cells were significantly increased(P<0.01).Sitagliptin decreased the viability and the expression of Col IV induced by AGEs in the mesangial cells in a dose-dependent manner.AGEs significantly inhibited the protein levels of beclin-1and p-AMPK,but significantly increased the protein level of p-p70S6K.Compared with AGE group,sitagliptin significantly reversed the above results in a dose-dependent manner.CONCLUSION:Autophagy may mediate the protective effect of sitagliptin on mesangial cells induced by AGEs.

关 键 词：西格列汀 自噬 系膜细胞 糖尿病肾病 Beclm-1 腺苷酸活化蛋白激酶 

分 类 号：R363.21[医药卫生—病理学] R587.1[医药卫生—基础医学]