SnoN对高糖诱导大鼠原代肾小管上皮细胞纤维连接蛋白合成的影响  

Effect of SnoN protein on high-glucose-induced expression of fibronectin in primary cultured rat renal tubular cells

作　　者：刘瑞霞[1] 郭兵[1] 王圆圆[1] 肖瑛[1] 石明隽[1] 张国忠[1] 

机构地区：[1]贵阳医学院病理生理学教研室,贵州贵阳550004

出　　处：《中国病理生理杂志》2011年第10期1931-1937,共7页Chinese Journal of Pathophysiology

基　　金：贵州省优秀科技教育人才省长专项基金资助项目(No.黔省专合字[2010]40号);贵阳市社会发展科技攻关资助项目(No.筑科农合同字第1-社-15号);贵阳医学院2009年研究生教育创新基地专项经费资助项目(No.B200903)

摘　　要：目的:观察核转录共抑制因子SnoN(Ski-related novel protein N)对高糖诱导大鼠原代肾小管上皮细胞(RTECs)纤维连接蛋白(FN)合成的影响。方法:原代培养大鼠RTECs,免疫荧光细胞化学和Western blot-ting检测培养细胞的上皮性钙黏蛋白、角蛋白-18和α-平滑肌肌动蛋白表达,鉴定培养细胞;正常糖(5.5 mmol/L)、高糖(25 mmol/L)和相同渗透压的甘露醇分别培养RTECs 30 min、2 h、12 h、24 h、48 h、72 h和96 h;SnoN siRNA转染RTECs,分为对照组、高糖组、control siRNA组和SnoN siRNA组;Western blotting和免疫细胞化学检测不同处理组RTECs中SnoN和FN蛋白的表达;RT-PCR检测FN mRNA。结果:与正常糖培养组相比,高糖刺激能抑制RTECs中SnoN蛋白表达,且呈时间依赖性;高糖刺激能使RTECs中FN蛋白和mRNA表达增加;与单纯高糖培养组相比,转染SnoN siRNA能进一步促进高糖诱导的RTECs中FN蛋白表达(P<0.05)。结论:SnoN表达减少参与了高糖诱导的RTECs中FN合成过程。AIM：To investigate the effect of Ski-related novel protein N（SnoN） on high-glucose-induced expression of fibronetin（FN） in primary cultured rat renal tubular cells（RTECs）.METHODS： The primary renal cells were cultured,and the cell types were indentified to be RTECs.The cells were divided into 3 groups： normal-glucose group（DMEM＋2% FBS）,high-glucose group（19.5 mmol/L D-glucose＋DMEM＋2% FBS） and high-osmotic group（19.5 mmol/L D-mannitol＋DMEM＋2% FBS）.The cells were harvested at 30 min,2 h,12 h,24 h,48 h,72 h and 96 h.SnoN expression in primary cultured RTECs was knocked down by RNA interference,then the cells were divided into 4 groups： normal-glucose group,high-glucose group,control siRNA group and SnoN siRNA group.The protein expressions of SnoN and FN in RTECs was examined by the methods of Western blotting,immunocytochemistry staining and immunofluorescence cytochemistry.RT-PCR was used to examine the mRNA expression of FN and SnoN.RESULTS： The RTECs constituted the major cell type of cultured cells.SnoN protein was decreased in a time-dependent manner in RTECs under high-glucose condition.The FN protein and mRNA levels raised in high-glucose group and sustained through entire experiment.Moderate reduction of SnoN in RTECs was observed by RNAi strategy,which greatly up-regulated the expression of FN（P〈0.05）.CONCLUSION： The down-regulation of SnoN participates high-glucose-induced expression of FN in RTECs.

关 键 词：高糖 肾小管上皮细胞 SNON 纤连蛋白类 

分 类 号：R363[医药卫生—病理学]