Effect of cysteine-rich protein 61 on oxidative stress in human kidney tubular epithelial cell line after anoxia / 中华肾脏病杂志

ABSTRACT

Objective To investigate the effect and mechanism of cysteine-rich protein 61 (Cyr61) on oxidative stress in human kidney tubular epithelial cell line after anoxia.Methods Human kidney tubular epithelial cell line (HK-2 cells) were divided into 5 groupscontrol group,Cyr61 group,MAPK inhibitor group (Cyr61 +PD98059),p38 inhibitor group (Cyr61 +SB203580) and PI3K inhibitor group (Cyr61+Wortmannin).Each group was pretreated for 12 h and then injured by anoxia.The cell viability was determined by MTT assay and the apoptosis rate of HK-2 cells was determined by flow-cytometry.The cellular ROS level was measured by spectro-fluorometry.The cellular superoxide dismutase (SOD) and catalase (CAT) were measured by nephelometry test.The expression of Nrf2 in HK-2 cells was detected by Western blotting.Results Anoxia enhanced the expression of ROS and Nrf2,decreased the expression of SOD and CAT significantly,meanwhile decreased HK-2 viability and increased HK-2 apoptosis (all P ＜ 0.05).Cyr61 increased the expression of p-Akt,Nrf2,SOD and CAT in HK-2,and decreased the expression of ROS,at the same time increased HK-2 viability and decreased HK-2 apoptosis (all P ＜ 0.05).Wortmannin inhibited the expression of p-Akt,Nrf2,SOD and CAT,meanwhile decreased HK-2 viability and increased HK-2 apoptosis (P ＜ 0.05).PD98059 and SB203580 had no affect on HK-2 compared to Cyr61 group (P＞0.05).Conclusions Cyr61 promotes the expression of Nrf2 through PI3K pathway in HK-2,which enhances the expression of SOD and CAT,and decreases the expression of ROS.Cyr61 exhibits protective effects on HK-2 cells injured by oxidative stress after anoxia.