Study Of Expression Regulation Of CaMK Ⅱ Affecting Hepatic Function And Cell Apoptosis After Liver Transplantation In Rats

Objective:This study is divided into 3 parts.Part 1:the establishment of the model of orthotopic liver transplantation in rats.Part 2:Lentiviral vectors carrying CaMK Ⅱ γ shRN A and CaMK Ⅱ and the same lentiviral empty vector as the former were transfected into the donor liver of rats to explore the influence of Ca2+/CaM/CaMK Ⅱ signaling pathway for liver cell apoptosis after liver transplantation in rats.Methods:1.The establishment of model of the orthotopic liver transplantation in rats.Model of the orthotopic liver transplantation in rats is built by operation training of two-person.2.The influences of the Ca2+/CaM/CaMK Ⅱ signaling pathway to apoptosis of liver transplantation in rats are explored.The healthy male SD rats were invided into 5 groups (A, B, C, D, E group) which have 20 rats for orthotopic liver transplantation.The donor liver were transfected with lentivirus vectors which carries CaMK Ⅱγ shRNA(A group), CaMK Ⅱ γ (B group), CaMK Ⅱγ shRN A empty lentivirus vector(C group), CaMKⅡγempty lentivirus vector (D group) and saline control group(E group) through portal vein when placed in preservation solution. The calcium ion condensity and the values of ALT,AST ofeach group of 1d,3d,7d,14d will be compared including the apoptosis degree of hepatocyte to study the influences of the Ca2+/CaM/CaMK Ⅱ signaling pathway to apoptosis of liver transplantation in rats.Results:1. The model of SD-SD orthotopic liver transplantation in rats is established successfully.2.The donor livers are transfected successfully by local perfusion in vitro. After selectively specific blocking CaMK Ⅱ (A group), its calcium condensity, values of ALT, AST and apoptosis rate of 7d are lower than others (B, C, D, E groups), the difference was statistically significant (P< 0.05); After up-regulated CaMK Ⅱ (B group), its calcium condensity, values of ALT, AST and apoptosis rate of 7d are higher than others (B, C, D, E groups), the difference was statistically significant (P<0.05); The differences of calcium condensity, values of ALT, AST and apoptosis rate of 7d between empty vector groups (C, D group) and control group (E group) was not statistically significant (P>0.05).Conclusions:1. The glucocorticoids may rise up the survive rate of rats after orthotopic liver transplantation.2.The selective blockage of the Ca2+/CaM/CaMKⅡ signaling pathway can obviously decline the occurance of ischemia-perfusion injury and apoptosis by reducing the calcium condensity after liver transplantation lighting up for treating hepatic insufficiency and ischemia-perfusion injury after liver transplantation in clinic.