| Liver transplantation has become the most effective treatment for the end stage of liver diseases. The donor shortage restricts the development of liver transplantation. The non-heart-beating donor(NHBD) liver graft suffers not only the warm ischemina injury but also the cold preservation and reperfusion injury. In the pathophysiological process, a number of cyokines, such as , TNF-α.et which are released with the activation of Kupffer cell, the obstruction of microcirculation, and the injury of liver cells and sinusoidal endothelial cells(SEC) become the major cause of primary non-function(PNF) of transplantating liver. How to take measures to reduce the ischemia-reperfusion and making good use of the NHBD is the studying hotspot. HSP as a kind of cellular endogenous protecting protein has many functions, such as molecular chaperone, antioxidating, cooperating with immunity, antiapoptosis and so on. And it can protect the liver ischemia-reperfusion injury obviously, before the graft harvest, increasing the expression of HSP of liver will lighten the liver ischemia-reperfusion injury. Objective: In this sdudy, we build the models of OLT of non-heart-beating donors of rats, induce the HSP by ischemic precondition, observe the effect of HSP70 on the liver graft ischemia-reperfusion injury at different time-point after non-heart-beating. The aim of this study is to investigate theoretical and experimental basis for using of the NHBD reasonably and effectively.Methods: seventy-six Wistar rats were randomly divided into two groups: IP (ischemic preconditioning) group and C (control) group. There are thirty-eight rats in each group. IP group: open the abdomen, sever the ligament around the liver, eliminate the collateral circulation, obstruct the hepatic portal for fifteen minutes by using the pringle's manoeuvre, the control group rats were only freed the hepatic portal. At 6h, 24h, 48h after the operation, graft samples of each group were taken out to detect the expression of HSP70. After 48 hours, the remainder rats were divided into 4 groups according to the ischemic preconditioning or not, and the warm ischemic time. The warm ischemic time was 45, 60 minutes respectively. All the rats were carried out orthotopic liver transplantation (OLT) as donors. At 1h after portal vein reperfusion ,eight rats of each group were executed, and blood samples were obtained to determine the levels of ALT and AST, grafts samples were obtained for optical microscope observation. And the survival rate of 1 week of the remainder rats were observed. One or two rats were selected randomly from the rats which survive for longer than one week, the other rats were executed to get the grafts samples for optical microscope observation. Detected the expression of HSP70 by Western blotting analysis; the levels of ALT and AST were determined by automatic biochemistry analysor; the liver samples were observed by optical microscope. During the operation cold ischemic time, non-hepatic time and surgical time of recipients were recorded.Results: 1. The HSP70 expression of IP group was low at 6h, and high at 24h ,48h. While there was almost no HSP70 expression in the C group at 24h, 48h.2. Cold ischemic time, non-hepatic time, surgical time among 4 groups were less discriminating (P>0.05). 3. At 1h after portal vein reperfusion, the levels of ALT and AST were significantly lower in the preconditioning groups (tN-45, tN-60) than in correspondingly non-preconditioning groups (N-45, N-60).4. At 1h after the reperfusion, in the N-45 group, some hepatocytes have eosinophilic degeneration, spotty necrosis, and vacuolar degeneration; in the N-60 group, there exist remarkable hepatic eosinophilic degeneration, cloudy swelling degeneration, spotty necrosis, vacuolar degeneration and inflammatory cell infiltration of portal area. While in the tN-45 and tN-60 groups, the structure of hepatic lobules are almost well, and the hepatocytes have no change like above. At one week after the operation, in the tN-45 and tN-60 group...
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