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本研究利用骨骼肌合成和分泌非肌性蛋白质,进而分泌入血并在细胞外发挥作用的原理,构建了一个由人巨细胞病毒早期增强子和促进子驱动的人apoEcDNA 表达质粒(pCMVapoE) ,通过阳离子脂质体介导转染原代培养的小鼠骨骼肌肌母细胞,ELISA和免疫组织化学检测均证实apoE 在肌源性细胞成功表达并分泌至培养液中。直接肌肉注射,pCMVapoE 表达载体也在小鼠骨骼肌中成功表达,并且预先经氯化钡诱导损伤可使其表达量明显增加。为进一步研究以骨骼肌为靶组织的动脉粥样硬化基因治疗提供了实验依据
In this study, we used the principle of skeletal muscle synthesis and secretion of non-muscle protein, and then secrete into the blood and play an extracellular role. We constructed a human apoEcDNA expression plasmid (pCMVapoE) driven by human cytomegalovirus early enhancers and promoters, Primary cultured mouse skeletal muscle myoblasts were transfected by cationic liposomes. ELISA and immunohistochemistry showed that apoE was successfully expressed in myogenic cells and secreted into the culture medium. By direct intramuscular injection, the pCMVapoE expression vector was also successfully expressed in mouse skeletal muscle and the expression level was significantly increased by barium chloride-induced injury in advance. This study provides the experimental basis for further research on the gene therapy of atherosclerosis with target tissue of skeletal muscle