人尿源性干细胞条件培养基对高糖诱导足细胞凋亡的影响  

Effects of Human Urine Derived Stem Cells on Human Podocytes Apoptosis Exposed to High Glucose

作　　者：姜珍珍[1] 汪年松[2] 汪泱[3] 刘四红[1] 王筱霞[2] 范瑛[2] 

机构地区：[1]苏州大学医学部,苏州215006 [2]上海交通大学附属第六人民医院肾脏内科,上海200233 [3]上海交通大学附属第六人民医院四肢显微外科研究所,上海200233

出　　处：《中国中西医结合肾病杂志》2014年第10期861-864,I0002,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然基金面上项目(No.81270824;81270814);上海市科委基金资助项目(No.11DZ1921904);上海市科委基础研究重大项目(No.11DJ1400101);上海市科委重大项目(No.11DZ1973103)

摘　　要：目的:通过正常人尿液提取干细胞,探讨尿源干细胞(human urine-derived stem cells,hUSCs)的条件培养基(conditioned medium,CM)对高糖所致人肾脏足细胞损伤的影响。方法:从健康男性尿液提取hUSCs,鉴定后进行体外培养,制备hUSCs-CM;体外培养人肾脏足细胞株,按培养条件不同,分为正常葡萄糖对照组(NG组)、高糖组(HG组)、甘露醇高渗对照组(MA组)及HG+hUSCs-CM不同剂量(1×,5×,10×)干预组,干预24 h后收集细胞,以RT-PCR检测足细胞骨架蛋白synaptopodin的表达;以Annexin V-FITC和PI双染方法,应用流式细胞仪观察各组足细胞的早期凋亡率。结果:HG组足细胞synaptopodin mRNA的表达较NG组降低(P<0.05),HG+hUSCs-CM(1×,5×,10×)组的足细胞synaptopodin mRNA的表达较HG组均增加(P<0.05);HG组足细胞凋亡率较NG组增高(P<0.05),hUSCs-CM干预组较HG组足细胞凋亡率均下降,并和hUSCs-CM具有剂量相关性(P<0.05)。结论:hUSCs-CM对高糖诱导的人足细胞凋亡及损伤具有保护作用。Objective：To extract stem cells through normal urine and to investigate the effects of conditioned medium（CM） from human urine derived stem cells（hUSCs）on podocyte lesion induced by high glucose. Methods：hUSCs were isolated from void-ed urine of healthy men. After identification and proliferation,hUSCs - CM was prepared. According to the different culture condi-tions,human podocytes were divided into normal glucose group,high permeability group,high glucose group（HG）,and HG ＋hUSCs - CM（1 × ,5 × ,10 × ）group. Real - time PCR was applied to detect the expression levels of synaptopodin,and human podocytes apoptosis in each group were examined by flow cytometry after cultured for 24 hours. Results：Compared with incubated in NG,the expression of synaptopodin mRNA in HG was found to decrease（vs. NG,P 〈 0. 05）. HUSCs - CM increased the expression of podocytic synaptopodin mRNA（vs. HG,P 〈 0. 05）. Early Podocytes apoptosis rate was mildly higher in HG group than in NG group（P 〈 0. 05）,hUSCs - CM reduced the early podocytes apoptosis in a dose - dependent manner（vs. HG,P 〈 0. 05）. Conclu-sion：hUSCs - CM prevents podocytic apoptosis and injury induced by HG,and may be a new therapeutic method to ameliorate the progress of DKD.

关 键 词：人尿源干细胞 条件培养基 人足细胞 凋亡 

分 类 号：R587.2[医药卫生—内分泌] R692[医药卫生—内科学]