腺病毒介导的心肌蛋白激酶Cε基因转移导致心功能损害

Adenovirus-mediated Protein Kinase Cε Gene Transfer to Myocardium of Mouse Lead to lmpaired cardiac function

目的 本文旨在研究腺病毒介导的成年小鼠心肌直接PKCε基因转移对左室收缩功能的影响。方法 使用标准方法建立表达PKCε基因的重组腺病毒载体。直接注射重组腺病毒到FVB/N和ICR小鼠心肌 ,对照鼠给予相同剂量空载腺病毒。Western免疫印迹测定PKCε蛋白质表达水平。用非开胸经颈总动脉插管的显微外科技术评价小鼠左室收缩功能。结果 与对照鼠相比基因转移鼠心肌转基因PKCε蛋白质表达水平增加近 4倍 ,基线左室最大收缩压、最大收缩速率 (dP/dt)、 -dP/dt明显降低 ,左室舒张末压、舒张压明显升高 (p <0 0 1) ,异丙肾上腺素激发后左室dP/dt剂量依赖的升高明显减弱 (p <0 0 1) ,PKCε基因转移鼠心脏 /体重比也较对照鼠明显增加 (p <0 0 1)。结论 腺病毒介导的、心肌直接的PKCε基因转移诱发了左室收缩功能的损害 ,导致了心肌肥厚和心衰。

Objective The present study aims to investigate the roles of the adenovirus-mediated PKCε gene transfer to myocardium of adult mouse on cardiac contractile function. Methods Recombinant adenovirus expressing PKCε gene was generated by using standard method. The PKCε recombinant adenovirus was directly injected to myocardium of FVB/N mice and ICR mice. Control mice were directly injected same dose of empty vector adenovirus. Expressive levels of PKCs protein were determined by Western immunoblotting. Left ventricular contraetile function was assessed by using in vivo closed-chest microscopic surgery method. Results PKCε gene transfer mice were associated with a 4-fold increase in PKCs transgenic protein compared to control mice. PKCε gene transfer mice exhibited decreased basal dP/dt, -dP/dt, left ventricle peak systolic pressure and enhanced left ventricle end diastolic pressure and diastolic pressure （p 〈 0.01 vs. control）. Dose-dependent increasing of dP/dt to isoproterenol in PKCε gene transfer mice was significantly attenuated when compared with control mice （p 〈 0.01）. And heart to body weight ratio in PKCεgene transfer mice was significantly increased （p 〈 0.01 vs. control）. Conclusion Adenovirus-mediated, myocardium-directed PKCs gene transfer impairs cardiac contractile function and leads to cardiac hypertroohy and failure.