白藜芦醇对嘌呤霉素氨基核苷诱导的足细胞氧化应激损伤的保护作用

Protective effect of resveratrol on oxidative stress injury of podocytes induced by puromycin aminonucleoside

目的探讨白藜芦醇(RSV)对嘌呤霉素氨基核苷(PAN)引起的足细胞氧化应激损伤的保护作用及可能机制。方法将小鼠肾脏足细胞系MPC5分为空白对照组(A组)、PAN处理组(B组)和PAN+RSV处理组(C组)。采用台盼蓝染色法和流式细胞术分别检测足细胞活力和凋亡情况,共聚焦显微镜下观察细胞骨架排列情况,DCFH-DA染色检测足细胞内活性氧(ROS)含量,比色法检测乳酸脱氢酶(LDH)和谷胱甘肽过氧化物酶(GSH-Px)活性,羟胺法检测超氧化物歧化酶(SOD)活性,硫代巴比妥酸法检测丙二醛(MDA)含量。结果与A组相比,B组小鼠足细胞活力、SOD和GSH-Px活性降低(P<0.05),足细胞凋亡以及ROS、LDH和MDA含量增加(P<0.05),足细胞骨架出现紊乱、重组和降解,并向边缘聚集;而C组经RSV干预后各项观察指标均得到有效改善(P<0.05)。结论 RSV可以部分逆转PAN诱导的足细胞损伤,其保护作用可能与抑制氧化应激、改善线粒体功能障碍相关。

Objective To investigate the protective effect and underlying mechanism of resveratrol(RSV) on oxidative stress injury of podocytes induced by puromycin aminonucleoside(PAN).Methods The mouse podocyte MPC5 was divided into three groups of A(blank control),B(treated with PAN 100 μg/mL) and C(treated with PAN 100 μg/mL and RSV 1 μg/mL).The cell viability and apoptosis were measured by trypan blue staining and flow cytometry, respectively.The change of cytoskeleton was observed under confocal microscopy.The content of reactive oxygen species(ROS) in podocytes was determined by DCFH-DA staining.The activities of LDH and GSH-Px were detected by colorimetric method.The activity of SOD and content of MDA were analyzed by hydroxylamine and thiobarbituric acid methods, respectively.Results Compared with group A,the viability of podocytes and activities of SOD and GSH-Px were decreased(P<0.05),the apoptosis of podocytes and contents of ROS,LDH and MDA were increased(P<0.05),and the disordered, restructured and degraded cytoskeleton was found in group B,which were all significantly improved after the intervention of RSV in group C(P<0.05).Conclusion RSV could partly reverse PAN-induced injury of podocytes, which may be related to the inhibition of oxidative stress and improvement of mitochondrial dysfunction.