Variation Of KIM-1 And NO In Ischemia-reperfusion Injuried NRK-52E Cells And Protection Of Cordyceps Sinensis On It

Objective To investigate the expression of NO and kidney injury molecule-1(Kim-1) in NRK-52E cells,which were induced ischemia-reperfusion injury by Antimycin A and observe the role of Cordyceps Sinensis on NRK-52E.Methods Culture cells, rat renal tubular epithelia cells(NRK-52E), were randomly divided into three groups: control, IRI, Cordyceps Sinensis + IRI group.CS+IRI group was pretreated for 72 hours by medium containing 40mg/L stock solution of cordyceps Sinensis,other two groups were cultured by blank medium for same time. After 72 hours, based on documented ways, were removed media and incubated with Antimycin A for 1 hour, and then recover the media to simulate the ischemia-reperfusion injury in vitro. On six time in the course (beginning, ischemia for 60min, ischemia 60min/reperfusion 10min, ischemia 60min/reperfusion 30min, ischemia 60min/reperfusion 60min, ischemia 60min/reperfusion 120min.), we detected the apoptosis ratio of cells by flow-cyto-meter and evaluated expression of Kim-1 by ELISA and reverse transcription-polymerase chain reaction (RT-PCR) and NO by Nitrate reductase method.Results Use the method to simulate the ischemia-reperfusion injury of NRK-52E by Antimycin A for 1 hour, we can observe the increasd expression of Kim-1(p<0.05). The apoptosis ratio significantly elevated in cells compared with the control group(p<0.05), the change above were time-dependent. The changes of expression of NO happened dynamically as the reperfusion time. Compared to model group, the Cordyceps Sinensis preincubated NRK-52E cells have showed apparente tolerance to ischemia-reperfusion injury, which manifestated depressed apoptosis ratio, decreased expression of NO and Kim-1.Conclusions Antimycin A can induce NRK-52E cells to ischemia-reperfusion injury, to apoptosis-based, accompanied by Early high expression of NO and Kim-1 in this process. The Cordyceps Sinensis can protect the NRK-52E cells from ischemia-reperfusion injury to some extent. The protect effection may link with the reduction of cell's apoptosis and downregulation of NO and Kim-1.