Protective Effects Of Salvia Miltiorrhiza Combined With Verapamil On Ischemia Reperfusion Hepatic Injury In Rats

ObjectiveTo study the protective effects of salvia miltiorrhiza combined with verapa- mil on ischemia-reperfusion hepatic injury in rats and to explore the mechanism of them on the ischemia-reperfusion hepatic injury so as to provide experiment- al evidence for them that was used to prevent and treat the ischemia-reperfus- ion hepatic injury in clinic.MethodsThe hepatic ischemia-reperfusion rat model was established by the method of Nauta. the rats were randomly divided into 5 groups: sham operation(S), is- chemia-reperfusion(I/R), salvia miltiorrhiza preconditioning(SM), verapamil preconditioning(V) and salvia miltiorrhiza combined with verapamil preconditi- oning(SV). At 2, 8, 24h after ischemia-reperfusion, immunohistochemistry was employed to detect the hepatic expressions of NF-κB and ICAM-1. Measureme- nt of hepatic tissue myeloperoxidase(MPO) concentration was used to reflect the neutrophilic infiltration of hepatic tissue. Hepatic tissue maleic dialdehyde(MDA) and superoxide dismutase(SOD) concentration were measured by the methods of the reagent box. And the injury severity of hepatic tissue was evaluated by the levels of serum alanine aminotransferase (ALT) and asparta- te aminotransferase(AST) and the examinations of light microscopy and elect- ron microscopy. ResultsThe NF-κB level at 2h(0.415±0.042)post-reperfusion was significantly higher in the I/R group. And it decreased gradually at 8h(0.310±0.031)and 24 h(0.244±0.034). And the ICAM-1 level increased at 2h(0.359±0.033)post-re- perfusion, peaked at 8h(0.450±0.034)and began to decrease at 24h(0.296±0.022). The changes of MDA, MPO, ALT and AST were similar to ICAM-1. The above-mentioned indicators at different time-points were lower in the SM, V and SV groups than those of the I/R group(P<0.05), but higher than the S group(P<0.05). the changes in ALT, MDA, MPO, NF-κB and ICAM-1 had a positive correlation(P<0.01). The SOD level at 2h(208.84±16.84)post-reperfusion decreased, it was lowest at 8h(184.80±12.96), and bagan to increase at 24h(202.78±14.70). SOD at different time-point was higher in the SM, V and SV groups than it of the I/R group(P<0.05), but lower than the S group(P<0.05) and it was much higher in the SV group than the SM and V groups. The chang- es in SOD had a negative correlation with ALT and MDA(P<0.01). Light and electron microscopy observations also showed significant differences in differe- nt groups apart from SM and V groups themselves.ConclusionsThe rat hepatic ischemia-reperfusion stimulates NF-κB activation, ICAM-1 expression and reduce the activity of SOD during hepatic ischemia-reperfusion injury process; Salvia miltiorrhiza and verapamil all can significantly reduce NF-κB activation, ICAM-1 expression and the activity of SOD after ischemia- reperfusion in hepatic tissue and effectively protect hepatic ischemia-reperfusi- on injury. In addition, salvia miltiorrhiza have collaborative effects combined with verapamil.