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

·论著·2012年11月第9卷第31期中国医药导报CHINA MEDICAL HERALD 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（MCs 1097），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（COL Ⅳ）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COL Ⅳ和FN 表达，PJ34显著抑制高糖诱导的COL Ⅳ和FN mRNA 过度表达（分别为高糖组的68.2%和54.7%（P <0.05）；COL Ⅳ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COL Ⅳ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COL Ⅳ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,Shenzhen 510082,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(COL Ⅳ),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 by 72.3%and 68.4%(P <0.05).PJ34reduced high glucose -induced upregulation of PARP-1mRNA and protein to 31.8%and 55.5%(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 by 1.77fold (P <0.05),compared with control cells.PJ34effectively inhibited the activa -tion of PARP-1by 32.2%(P <0.05),compared with the cells treated with high glucose alone.Also,high glucose in -creased COL Ⅳand FN https://www.360docs.net/doc/5011918884.html,pared with high glucose stimulation group,PJ34suppressed high glucose -induced upregulation of COL Ⅳand FN mRNA by 31.8%and 45.3%,respectively (P <0.05).The changes in COL Ⅳand FN were confirmed at protein level.the inhibition rate of COL Ⅳand FN protein expression were 54.0%and 66.6%,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