胆固醇逆转运相关蛋白基因在骨骼肌的表达  

Expression of Reverse Cholesterol Transport Pathway Associated Protein Genes in Skeletal Muscle

作　　者：范乐明[1] 张慧[1] 陈琪[1] 魏恩会[1] 蔡海江[1] 

机构地区：[1]南京医科大学动脉粥样硬化研究中心,南京210029

出　　处：《生物化学与生物物理学报》2000年第2期109-114,共6页

基　　金：江苏省科委应用基础基金资助项目!No .BJ980 87&&

摘　　要：构建载脂蛋白AI(apoAI)、载脂蛋白E(apoE)和卵磷脂胆固醇酰基转移酶 (LCAT)的病毒或非病毒载体 ,分别转染离体成肌细胞或直接注入小鼠骨骼肌 ,观察其表达程度及分泌入血等情况 ,探讨借用骨骼肌异源表达这些在胆固醇逆转运中起关键作用的重要候选基因 ,进而发展一种简易安全基因治疗方法的可能性。结果显示 ,质粒表达载体 pCMVapoE3和重组腺病毒载体Ad RSV apoAI在原代培养小鼠成肌细胞和成肌细胞株C2C12成功表达并分泌至培养液 ;直接肌肉注射重组病毒使apoAI在小鼠骨骼肌表达并分泌进入血液循环 ,持续 3 0d。LCAT的质粒表达载体和重组腺辅助病毒 (AAV)质粒DNA均可使C2C12和 2 93细胞表达出有功能的LCAT ;重组AAV质粒DNA的表达效果比常规质粒表达载体高 2～ 5倍。Viral and nonviral vectors containing apoA I, apoE or LCAT genes were constructed and transfected into myogenic cells in vitro or injected directly into mouse skeletal muscle. The expression efficiencies of these vectors were assaied to investigate the possibility of ectopic expression of these genes in skeletal muscle and to develop a safe and convenient gene therapy method for atherosclerosis. The results showed that the primary cultured mouse myoblasts, C2C12 cells transfected with pCMV apoE 3 expressed human apoE3 successfully and the expressed product was secreted into the medium. Mouse skeletal muscle efficiently expressed apoE3 in vivo after direct plasmid injection. The expression level of Ad RSV apoA I in primary cultured mouse myoblasts was correlated with virus titer. Human apoAI was synthesized in mouse skeletal muscle by direct injection of recombinant virus and was secreted into blood continuously up to 30 days. Functional LCAT was expressed by C2C12 and 293 cells transfected with conventional vector or recombinant AAV plasmid DNA. The expression efficiency of recombinant AAV plasmid DNA was 2—5 times higher than that of conventional plasmid vector. The above results provided experimental data for further studying and developing a gene therapy method for atherosclerosis by enhancement of reverse cholesterol transport using skeletal muscle as target.

关 键 词：胆固醇逆转运 动脉粥样硬化 基因治疗 高脂血症 

分 类 号：Q786[生物学—分子生物学]