CAJ | 학술논문

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。
목적：탐토인제대간충질간세포（HUC?MSCs）대고당유도적족세포조망적영향급기궤제。방법소서족세포（MPC5）세포피분위6조：（1）정상당농도조（NG）；（2）고당조（HG）；（3）감로순조（NG+MA）；（4）HUC?MSCs공배양조（HUC?MSCs）；（5）HUC?MSCs+중조인간세포생장인자（HGF）중화항체조（HGF?NtAb）；（6）중조인간세포생장인자조（rhHGF）。자격72 h후，류식세포술검측족세포조망솔；Western인적법검측조망표지단백다취ADP핵당취합매（PARP）、세포조망억제인자Bcl?2적표체；세포면역형광법검측족세포상관분자적표체급배렬；Hoechst염색법검측족세포조망수평；ELISA법검측HUC?MSCs분비HGF수평。결과여NG조비교，HG조MPC5세포조망표지단백PARP적표체증다，Bcl?2표체감소，처리72 h조조간비교차이유통계학의의（P﹤0.05）；족세포상관분자podoplanin표체감소，족세포조망솔증고（P﹤0.05）。여HG조비교，HUC?MSCs조，rhHGF조PARP적표체감소、Bcl?2표체증가（P﹤0.05）， podoplanin표체정상，족세포조망솔감저（P﹤0.05）。여HUC?MSCs조비교，가입HGF중화항체후PARP적표체증다，Bcl?2표체감소（P﹤0.05），podoplanin표체감소，족세포조망솔증고（P﹤0.05）。결론고당자격가유도족세포발생조망화손상，HUC?MSCs공배양가이개선고당유도적족세포조망화손상，HUC?MSC가능통과분비HGF완해고당유도적족세포조망화손상。
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.