中介素预处理对NRK-52E细胞缺氧/复氧损伤的保护作用及其机制OACSTPCD

Objective To explore the protective effect of intermedin(IMD)against rat renal tubular epithelial cell(NRK-52E)injury induced by hypoxia/reoxygenation(H/R).Methods The NRK-52E cells were randomly divided into normal control(NC)group,CoCl2 group and IMD+CoCl2 group.H/R injury models were induced by CoCl2(1.5×10-4 mol/L)for 24 h.The cells in IMD+CoCl2 group were incubated with IMD 20 ng/mL for 4 h,and then cultured with 20 ng/mL IMD and 1.5×10-4 mol/L CoCl2 for 24 h.Flow cytometry was used to access the apoptosis rate of cells,reactive oxygen species(ROS)generation was detected by DCFH-DA probe,mitochon-drial membrane potential(MMP)was detected by JC-1 probe.Real time PCR was employed to detect the mRNA expression levels of Bcl-2,Caspase-3,HO-1 and eNOS.Western blot was employed to detect the protein expression levels of Bcl-2,Caspase-3,HO-1 and eNOS.Results Compared with NC group,the cell apoptosis rate was significantly increased in CoCl2 group(P<0.01),the intracellu-lar ROS level was significantly increased and MMP was significantly decreased,the mRNA and protein expression levels of Caspase-3 were significantly increased(P<0.01),and the protein expression of Bcl-2 was significantly decreased(P<0.05).Compared with CoCl2 group,the cell apoptosis rate was significantly decreased in IMD+CoCl2 group(P<0.01),ROS level was decreased while MMP was increased,the mRNA and protein expressions of Caspase-3 were significantly decreased(P<0.05),and the mRNA and protein expres-sion levels of Bcl-2,HO-1 and eNOS were significantly increased(P<0.05).Conclusion IMD could ameliorate the hypoxia/reoxy-genation-induced injury in NRK-52E cells by inhibiting cell apoptosis and oxidative stress,and increasing the expression levels of Bcl-2,eNOS and HO-1.