多聚二磷酸腺苷(ADP)核糖聚合酶-1(PARP-1)在高糖致大鼠肾脏系膜细胞细胞外基质沉积中的作用

高血糖作为糖尿病的最基本生化特性,通过多种机制引起糖尿病肾病（diabetic nephropathy，DN），其中氧化应激是一个重要损伤机制。而有研究发现高血糖介导的ROS可以激活多聚ADP核糖聚合酶[poly（ADP-ribose）polymerase，PARP][1]。PARP是一种存在于除酵母外所有真核细胞核内对DNA断裂敏感的蛋白酶，目前其在DN发病机制中的作用尚未完全阐明。DN时，肾小球系膜细胞是多种致病因子作用的主要靶细胞，主要表现为肾小球的肥大、细胞外基质的堆积、肾小

多聚二磷酸腺苷（ADP）核糖聚合酶-1（PARP-1）在高糖致大鼠肾脏系膜细胞细胞外基质沉积中的作用

朱恒梅祝胜郎▲陈结慧叶玲蒋莹李向阳

广东省深圳市第六人民医院肾内科，广东深圳510082

[摘要]目的探讨多聚二磷酸腺苷（ADP）核糖聚合酶-1（PARP-1）在高糖（25mM）致大鼠肾脏系膜细胞细胞外基质沉积中的作用。方法①体外培养大鼠肾脏系膜细胞株（MCs1097），使用高糖（25mM）处理大鼠肾脏系膜细胞，部分实验组中应用PARP-1抑制剂PJ34（3×10-6M）进行干预处理。②RT-PCR和Western blot检测PARP-1、纤维粘连蛋白（FN）、胶原Ⅳ（COLⅣ）的mRNA及蛋白表达。③高通量比色测定法检测PARP-1的活性。结果①高糖显著诱导大鼠肾脏系膜细胞PARP-1mRNA和蛋白的表达，分别较低糖对照组增加72.3%和68.4%（P<0.05），PJ34可明显抑制高糖诱导的PARP-1mRNA和蛋白的过度表达，mRNA和蛋白分别为高糖组的31.8%和55.5%（P<

0.05）。同时，高糖显著诱导大鼠肾脏系膜细胞PARP-1激活，高糖组为低糖对照组活性的1.77倍（P<0.05），PJ34

可显著抑制高糖诱导的PARP激活，活性降低为高糖组的67.8%（P<0.05）。②高糖显著增加COLⅣ和FN表达，PJ34显著抑制高糖诱导的COLⅣ和FN mRNA过度表达（分别为高糖组的68.2%和54.7%（P<0.05）；COLⅣ和FN蛋白水平的变化与RNA水平变化相一致，PJ34可显著抑制高糖诱导的COLⅣ和FN蛋白表达（分别为高糖组的54.0%及66.6%（P<0.05）。结论高糖可以诱导培养的大鼠肾脏系膜细胞内PARP激活，PARP-1表达增加，PJ-34预处理可以明显降低高糖诱导的大鼠肾脏系膜细胞内PARP的激活以及下游FN及COLⅣ的高表达，提示PARP1参与了肾脏系膜细胞细胞外基质沉积的发生发展过程。

[关键词]系膜细胞；PARP；高血糖；细胞外基质

[中图分类号]R587.2[文献标识码]A[文章编号]1673-7210（2012）11（a）-0008-04

Poly(ADP-ribose)polymerase-1mediates high glucose-induced extracel-lular matrix accumulation in rat mesangial cells

ZHU Hengmei ZHU Shenglang▲CHEN Jiehui YE Ling JIANG Ying LI Xiangyang

Department of Nephrology,the Sixth People's Hospital of Shenzhen City,Guangdong Province,Shenzhen510082,China [Abstract]Objective To investigate the role of Poly(ADP-ribose)Polymerase-1(PARP-1)in high glucose-induced accu-mulation of extracellular matrix(ECM)in rat mesangial cells(RMCs).Methods RMCs were treated with or without high glucose(25mmol/L).In some experimental groups,cells were pre-treated with PARP-1inhibitor PJ34(3×10-6mol/L).

RT-PCR was employed to detect the mRNA expression for ADP-ribose polymerase-1(PARP-1),fibronectin(FN).collagen Ⅳ(COLⅣ),and all those proteins expression were examined by Western blot.The activity of PARP-1was examined by Colorimetric assay.Results high glucose significantly stimulated both PARP-1mRNA and protein overexpression in RM-Cs.the mRNA and protein expression of PARP-1were up-regulated by72.3%and68.4%(P<0.05).PJ34reduced high glucose-induced upregulation of PARP-1mRNA and protein to31.8%and55.5%(P<0.05),compared with high glucose stimulation group.At the same time,high glucose obviously stimulated PARP-1activation,the PARP-1activity of high glucose group was upregulated by1.77fold(P<0.05),compared with control cells.PJ34effectively inhibited the activa-tion of PARP-1by32.2%(P<0.05),compared with the cells treated with high glucose alone.Also,high glucose in-creased COLⅣand FN pared with high glucose stimulation group,PJ34suppressed high glucose-induced upregulation of COLⅣand FN mRNA by31.8%and45.3%,respectively(P<0.05).The changes in COLⅣand FN were confirmed at protein level.the inhibition rate of COLⅣand FN protein expression were54.0%and66.6%,respec-tively,compared with high glucose stimulation group(P<0.05).Conclusion Our data suggest that PARP-1mediates high glucose-induced accumulation of ECM in rat mesangial cells and thus may represent a potential therapeutic target in the management of glomerular disease.

[Key words]Mesangial cells;PARP;Extracellular matrix

[基金项目]2011广东省医学科研立项（编号：A2011570）。2011年广东省深

圳市科技局科技立项（编号：201102134）。广东省深圳市南山区科技局基

金（编号：南卫2011004）。

▲通讯作者

8

中国医药导报CHINA MEDICAL HERALD