人脐带间充质干细胞共培养改善高糖诱导的足细胞凋亡  

Human umbilical cord mesenchymal stem cells co-culture ameliorates podocytic apoptosis：a possible role of HGF

作　　者：齐文文[1] 吕莎莎[1] 柳刚[1] 程静[1] 宋燕[1] 明彤彤 关广聚[1] 

机构地区：[1]山东大学第二医院肾内科山东大学肾脏病研究所,济南250033

出　　处：《中华肾脏病杂志》2014年第12期933-938,共6页Chinese Journal of Nephrology

基　　金：山东省科技发展计划基金（2013GSF12104）

摘　　要：目的：探讨人脐带间充质干细胞（HUC-MSCs）对高糖诱导的足细胞凋亡的影响及其机制。方法小鼠足细胞（MPC5）细胞被分为6组：（1）正常糖浓度组（NG）；（2）高糖组（HG）；（3）甘露醇组（NG＋MA）；（4）HUC-MSCs共培养组（HUC-MSCs）；（5）HUC-MSCs＋重组人肝细胞生长因子（HGF）中和抗体组（HGF-NtAb）；（6）重组人肝细胞生长因子组（rhHGF）。刺激72 h后，流式细胞术检测足细胞凋亡率；Western印迹法检测凋亡标志蛋白多聚ADP核糖聚合酶（PARP）、细胞凋亡抑制因子Bcl-2的表达；细胞免疫荧光法检测足细胞相关分子的表达及排列；Hoechst染色法检测足细胞凋亡水平；ELISA法检测HUC-MSCs分泌HGF水平。结果与NG组比较，HG组MPC5细胞凋亡标志蛋白PARP的表达增多，Bcl-2表达减少，处理72 h组组间比较差异有统计学意义（P﹤0.05）；足细胞相关分子podoplanin表达减少，足细胞凋亡率增高（P﹤0.05）。与HG组比较，HUC-MSCs组，rhHGF组PARP的表达减少、Bcl-2表达增加（P﹤0.05）， podoplanin表达正常，足细胞凋亡率减低（P﹤0.05）。与HUC-MSCs组比较，加入HGF中和抗体后PARP的表达增多，Bcl-2表达减少（P﹤0.05），podoplanin表达减少，足细胞凋亡率增高（P﹤0.05）。结论高糖刺激可诱导足细胞发生凋亡和损伤，HUC-MSCs共培养可以改善高糖诱导的足细胞凋亡和损伤，HUC-MSC可能通过分泌HGF缓解高糖诱导的足细胞凋亡和损伤。Objective To explore the effects of human umbilical cord mesenchymal stem cells （HUC-MSCs） on podocytic apoptosis and injury induced by high glucose （HG） and the underlying mechanisms. Methods Podocytes were divided into six groups according to treatment： ⑴ normal glucose group （NG）;⑵high glucose group （HG）;⑶mannitol control group （NG＋Ma）;⑷HUC-MSC co-culture group （HUC-MSCs）; ⑸ recombinant human hepatocyte growth factor treatment group （rhHGF）;⑹ neutralizing antibody group（HGF-NtAb）. Cytometry and Hoechst staining were used to detect the apoptosis rates. Western blot was used to measure the ratio of active PARP to total PARP and the level of Bcl-2. Immunofluorescence was used to study podocytic apoptosis and injury. Neutralizing antibody （NtAb） was used to block its function and the recombinant cytokine was added to induce its function. Results High glucose induced podocytic apoptosis in a time-dependent manner, HUC-MSCs co-culture decreased the podocytic apoptosis rate and the expression of PARP （all P﹤0.05）, increased the expression of Bcl-2, prevented the reduced expression and maintained the normal arrangement of podocytic podoplanin. The rhHGF prevented podocytic apoptosis and injury similarly to HUC-MSCs, the beneficial effect of HUC-MSC decreased when blockade of HGF. Conclusions HUC-MSCs co-culture ameliorates podocytic apoptosis and injure induced by HG, probably through secreting soluble HGF.

关 键 词：糖尿病肾病 干细胞 间充质 足细胞 凋亡 肝细胞生长因子 

分 类 号：R587.2[医药卫生—内分泌]