BackgroundDonor graft preservation is one of the most important issues in liver transplantation. Since a good graft is the precondition for the satisfactory postoperative liver function , scientists have been working for decades to find out a high efficacy and low cost drug for preservation. Since calcium overload after cold preservation causes serous hepatocyte injury, calcium channel blockers have been suggested one of choice for the preservation of liver graft from injury.Hanfangchin A, also called Tetrandrine(TET), is a kind of bisbenzylisoquinoline alkaloid refined from Fourstamen stephania root. It is a calcium blocker made in China which is pharmalogically similar to verapamil. Studies on freshly isolated hepatocytes has revealed that TET can significantly improve cell function, reduce pathological injury, inhibit the production of extracellular matrix and free radicals and promote the production of SOD. Although its effects on liver graft after cold preservation have not been reported, the evidences mentioned above suggest that TET may play a role. ObjectiveThe present study is to investigate whether TET has protective effect on rat donor graft by the pretreatment of TET on donor liver and cold perfusion with different solutions, including TET-HCA, HCA and UW solutions and what the mechanisms are, evaluating the probability of TET in the application of clinical liver transplantation. Part One the preservation for liver graft with TET1. Material and methodSD rats were pretreated by intraperitoneal injection or intraportal perfusion of TET containing solutions. Nimodipine solution and Ringer lactate solution were used as controls. Afterwards, free calcium, ATP in hepatocytes and ALT, LDH in preserved solutions were monitored and pathological changes of the preserved liver were observed.2. Resultsa. In the TET intraperitoneally injected group, free calcium concentration greatly decreased, energy metabolism significantly improved, accompanied by declining of ALT and LDH and mild pathological alterations, suggesting that TET may decrease calcium overload and energy consumption during cold preservation, thus ameliorating liver injury and enhance viability of donor liver cells.b. There were no significant changes between TET and nimodipine groups in intracellular free calcium concentration, energy metabolism, ALT, LDH concentration and liver pathological alterations, suggesting that preservation effect of TET is similar to nimodipine.c. Hepatocellular free calcium concentration decreased more significantly in TET intraportal injection group than that in intraperitoneal injection group. As compared with TET intraperitoneal group, ALT, LDH decreased more significantly and there was a milder pathological alteration in intraportal group, demonstrating that pretreatment of donor liver by intraportal injection is a better way to preserve donor liver graft.Part Two the comparsion of preservating validity to TET and UW solution for liver graft.1. Material and methodSD rats were cold preserved 2 hrs by various solutions, including TET-Ringer, TET-HCA, HCA and UW solutions. Afterwards, hepatocytes ATP and ALT, LDH in solutions were monitored and liver pathological changes observed.2. Resultsa. No significant changes were observed in energy metabolism , ALT , LDH pathological findings between TET-Ringer and TET-HCA groups, suggesting similar effects between the two groups by 2 hr cold preservation.b. Energy metabolism in TET-HCA group was much improved as compared with that in HCA group, accompanied by more significant decrease in ALT and LDH concentration and milder pathological changes, suggesting better preservation effect of TET-HCA compared to that of HCA group.c. No significant changes were observed in energy metabolism , ALT , LDH pathological findings between TET-HCA and UW groups, suggesting similar effects between the two groups by 2 hr col