摘要
该文探究了C1q/肿瘤坏死因子相关蛋白-3(C1q/TNF-related protein 3,CTRP3)在高糖条件下肾小管细胞胆固醇转运中的作用及机制。体外培养人肾小管上皮细胞HK-2,随机分为正常糖对照组(NG)、正常糖+CTRP3干预组(NG+CT)、高糖培养组(HG)、高糖培养+CTRP3干预组(HG+CT)、高糖培养+CTRP3干预组+siRNA转染组(HG+CT+siRNA)和高糖培养+CTRP3干预组+Sirt1 siRNA转染组(HG+CT+siSirt1)。试剂盒检测细胞内胆固醇含量及胆固醇流出情况;Filipin染色观察细胞内胆固醇蓄积情况;试剂盒检测Sirt1酶活性;Western blot检测CTRP3、Sirt1、ABCA1及LXRα的蛋白表达;实时定量PCR检测CTRP3的mRNA表达。结果显示,重组CTRP3蛋白干预能够抑制高糖条件下HK-2细胞胆固醇蓄积,上调ABCA1及LXRα的表达从而促进胆固醇外流;同时增强Sirt1的蛋白表达及活性;应用Sirt1 siRNA抑制Sirt1的表达后CTRP3的上述调控作用均消失了。以上结果提示,CTRP3对高糖条件下的肾小管细胞的保护作用,可能是部分通过调控Sirt1的表达促进高糖条件下肾小管细胞的胆固醇外流实现的。
The purpose of this study is to investigate the role and molecular mechanisms of CTRP3 on cholesterol transport in renal tubular cells under high glucose condition.Cultured HK-2 cells were randomly divided into normal glucose group(NG),NG plus CTRP3 group(NG+CT),high glucose(HG),HG plus CTRP3 group(HG+CT),HG plus CTRP3 treatment plus siRNA transfection group(HG+CT+siRNA),HG plus CTRP3 treatment plus Sirt1 siRNA transfection group(HG+CT+siSirt1).Cholesterol content and efflux levels in different groups were detected using cholesterol detection kits according to instructions of the manufacturer,and cholesterol accumulation was detected by Filipin staining.The activity of Sirt1 enzyme was measured using commercial detection kit.Western blot was used to detect the protein expression of CTRP3,Sirt1,LXRα,and ABCA1 in HK-2 cells.The level of CTRP3 mRNA was detected by Real-time PCR.Recombinant protein CTRP3 treatment significantly inhibited the HG-induced cholesterol accumulation and promoted the level of cholesterol efflux in HK-2 cells.In addition,down-regulation of CTRP3,Sirt1,LXRα,and ABCA1 protein expression and Sirt1 activity induced by HG were markedly prevented by treatment with recombinant protein CTRP3.After the application of Sirt1 siRNA to inhibit the expression of Sirt1,the above regulatory effects of CTRP3 on HK-2 cells disappeared.These findings indicated that CTRP3 had protective effects on HG-induced cholesterol accumulation in HK-2 cells.The mechanism of role performed by CTRP3 may include,at least in part regulating the expression of Sirt1 and promoting the cholesterol efflux.
作者
梁子辉
朱艳
任韫卓
吴明
韩伟霞
张国宇
胡越
杜春阳
LIANG Zihui;ZHU Yan;REN Yunzhuo;WU Ming;HAN Weixia;ZHANG Guoyu;HU Yue;DU Chunyang(Hebei Medical University,Department of General Theory of surgery and Surgical Science,Shijiangzhuang,050017,China;Hebei Medical University,Department of Electron Microscope,Shijiazhuang,050017,China;Hebei Medical University,Department of Pathology,Shijiazhuang 050017,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2019年第11期2108-2115,共8页
Chinese Journal of Cell Biology
基金
河北省教育厅重点研发计划项目(批准号:ZD2018007)
河北省重点研发计划项目(批准号:18277748D)
河北医科大学大学生创新实验项目(批准号:USIP2018183、USIP2018151)资助的课题.
作者简介
通讯作者:杜春阳,Tel:0311-86265724,E-mail:duchunyang55@163.com
