Experimental Study On Interventing Factors During Hepatic Ischemic-reperfusion Injury

Objective To observe the protective effect of verapamil and Shengmai injection on ischemic-reperfusion injury in rat liver.Methods A rat model of liver ischemic-reperfusion injury was established. 40 healthy female Wistar rats were randomly divided into 4 groups as follows: Sham operation group(A group), ischemic-reperfusion group(B group), verapamil group(C group), pretreated with verapamil by caudal vein injection 10 minutes before ischemia, Shengmai injection group(D group), pretreated with shengmai injection by intraperitoneal injection 3 days before ischemia. Ischemia of the left lobe was created by total inflow occlusion for 30 minutes followed by 24-hour perfusion. Then liver tissues were obtained for light microscopy study by formalin-fixed, paraffin-embedded, slices cut and HE staining. The heat shock protein70(HSP70) expression level was determined by western blotting. Malondialdehyde(MDA) was measured by TBA colorimetric method. Serum levels of tumor necrosis factor alpha(TNF- α ) was determined using radioimmunoassay kit , and alanine aminotrasferase(ALT), aspartate aminotrasferase (AST) were measured by full-automatic biochemical analyzer.Result (1) In A group,there were the lest TNF- α , MDA ,ALT, AST changes,as well as lowest HSP70 expression level, and normal liver tissue morphologic changes.(2)In B group, after ischemic-reperfusion, there were more TNF- α , MDA .ALT, AST changes as well as higher HSP70 expression level, and the morphologic injury changes of liver tissues more obvious than in the other three groups.(3) ALT was 109.13±18.91,131.75±21.23(U/L) separately in C group and D group remarkably reduced than 183.89±27.90 in B group(q=10.08,7.03, P<0.01), and there was significantly different between C group and D goup(q=2.97, P<0.05);AST was 269.50±51.95(U/L)(C group),281.63±52.54(U/L)(D group), remarkably reduced than 331.11±49.51(U/L) in B group(q=3.82,3.07, P<0.05) , but no significant difference was found between C group and D group(q=0.73,P>0.05);TNF- α was 143.69±16.94(pg/ml) (C group),146.141±4.39(pg/ml) (D group), significantly reduced than 177.56±24.50(pg/ml) in B goup(q=3.72,3.45, P<0.05), but no significant difference was found between C group and D group(q=0.26,P>0.05);MDA was 1.11±0.10(nmol/ml) (C group),1.15±0.08(nmol/ml) (D group) significantly reduced than 1.310±0.22(nmol/ml) in B group(q=3.61.2.89, P<0.05), but no significant difference was found between C group and D group(q=0.70, P>0.05);HSP 70 expression levels were obviously lower than that in B group, while a little less in Cgroup than in D group;the liver cellular edema necrosis and inflammatory infiltrationwere effectively alleviated.Conclusion After ischemic reperfusion,the morphologic injury changes of rat livertissues were obvious.The mechanism might be related to calcium overload andgeneration of oxygen free radicals;Verapamil and shengmai injection administeredbefore ischemia could effectively protect the rat liver from ischemic-reperfusioninjury.