Effect Of CaM/CaMK Ⅱ On Mitochondria Damage、 Apoptosis In Rat Liver Transplantation

Objective:By adjusting CaMK Ⅱ level, to explore the effect of CaM/CaMK Ⅱ in liver cell mitochondria damage、apoptosis in rats after liver transplantation.Methods:Healthy male SD rats were randomly divided into five groups (A、B、C、 D、E),each set of 10 pairs used to establishing " Kamada two cuff method " rat orthotopic liver transplantation model. Lentiviral expression vectors of CaMK Ⅱγ shRNA (A)、CaMK Ⅱγ (B) and Lentiviral empty vectors of CaMK Ⅱγ shRNA (C)、 CaMK Ⅱγ (D) are used to transfection graft by portal vein infusion.The control group (E) is infused normal saline. At the time of 1d、3d、7d、14d after surgery, to detect protein levels of CaM、CaMK Ⅱ、 cytochrome C、 ALF by methord of Western bolt to detect levels of CaM、CaMK Ⅱ mRNA by QRT-PCR and Cell apoptosis was detected by flow cytometry.Results:1、successfully established " Kamada two cuff method " of orthotopic liver transplantation model.2、A group at each time point evel of CaMK Ⅱ、 CytC、 ALF are higher, but CaM is lower, differences were statistically significant (P< 0.05). B group at each time point evel of CaMK Ⅱ、CytC、ALF are higher, but CaM is lower, differences were statistically significant (P< 0.05). A group at each time point evel of CaM mRNA is lower, CaMK Ⅱ mRNA is lower, differences were statistically significant (P< 0.05). B group at each time point evel of CaM mRNA is lower, but CaMK Ⅱ mRNA is higher, differences were statistically significant (P< 0.05). C、 D、E group was not statistically significant (P> 0.05). Cell apoptosis was detected by flow cytometry levels:the lowest group A, the highest group B, C、D、E group had no significant difference.Conclusion:By downregulation CaMK Ⅱ level in theCaM/CaMK Ⅱ signaling pathways, Cyt C, AIF protein levels in liver cells can be reduced and so the hepatic mitochondria injury、apoptosis, after liver transplantation in rat.