CaMKⅡ表达调控对大鼠肝移植术后肝功能的影响

Effect of CaMKⅡ expression regulation on liver function after liver transplantation in rats

目的探讨钙调素依赖蛋白激酶家族(calmodulin-dependent kinases casades,CaMK)Ⅱ表达调控对大鼠肝移植术后肝功能的影响。方法采用经典二袖套法建立SD至SD大鼠同种异体原位肝移植模型,并构建CaMKⅡγ蛋白和CaMKⅡγsh RNA的慢病毒表达体系。分别将表达CaMKⅡγsh RNA的慢病毒载体及表达CaMKⅡγ蛋白的慢病毒载体灌注至肝移植术后大鼠体内,同时予以灌注相应的空载体及生理盐水形成对照,分别于术后不同时间点取大鼠下腔静脉血测定其血清ALT及AST水平。结果灌注表达CaMKⅡγsh RNA的慢病毒载体的移植术后大鼠血清ALT及AST降低(P<0.05);灌注表达CaMKⅡγ蛋白的慢病毒载体的移植术后大鼠血清ALT及AST升高(P<0.05);灌注空白载体及生理盐水的移植术后大鼠血清ALT及AST均无明显差异(P>0.05)。结论特异性阻断CaMKⅡ信号通路可有效降低大鼠肝移植术后的血清ALT及AST水平;而增强的CaMKⅡ信号通路则升高大鼠肝移植术后的血清ALT和AST水平。

Objective To investigate the effect on expression regulation of calmodulin-dependent kinases casades （CaMK） II on liver function after liver transplantation in rats. Methods Allogeneic orthotopic liver transplantation model was established by using the classic two-cuff method. The lentiviral expression systems of CaMK II γ protein and CaMK II y shRNA were constructed. The lentiviral vector expressing CaMK II γ shRNA and the lentiviral vector expressing CaMK IIγ protein were perfused into the rat after liver transplantation respectively, and the corresponding blank vector and normal saline were perfused into the control group at the same time. The serum levels of ALT and AST were measured at different time points of inferior vena cava blood in rats. Results The serum ALT and AST levels were debased in the after transplantation rats whose lentiviral vector expressing CaMK II γ shRNA （P〈0.05）. The serum ALT and AST levels were raised in the after transplantation rats whose lentiviral vector expressing CaMK II y protein （P〈0.05）. There were no significant difference of serum ALT and AST levels between the blank control group and the saline group （P〉0.05）. Conclusion Specific blocking of CaMK II signaling pathway can effectively reduce the serum ALT and AST levels after liver transplantation in rats, and enhanced CaMK II signaling pathway increases the serum ALT and AST levels after liver transplantation in rats.