分泌型αFGF基因转染鼠心肌细胞在大鼠体内外表达的研究

Study of secretory αFGF gene transfected to cardiomyocytes in vitroand in vivo

  • 摘要: 目的:观察分泌型αFGF真核表达载体转染大鼠心肌细胞后αFGF在体内外的分泌表达及其对宿主心肌血管形成作用的影响。方法:构建分泌型αFGF真核表达载体(pCDNA 3.1+SP αFGF),转染培养大鼠的心肌细胞,体外检测αFGF的分泌表达;将转染细胞移植入同种大鼠心肌,检测αFGF在体内的表达及受体心肌的血管生成。结果:硝酸纤维素膜点杂交和ELISA显示,αFGF在转染24-96h细胞培养上清中有较高的表达;接受转染心肌细胞移植96 h后免疫组化检查发现αFGF在受体心肌内得到分泌表达,20 d后组织切片发现移植心肌细胞周围的心肌中有较多的新生毛细血管。结论:转染分泌型αFGF真核表达载体的心肌细胞在体内外能分泌表达αFGF;通过心肌细胞移植,分泌的αFGF在受体心肌组织中有较强的血管生成作用。

     

    Abstract: Objective: To investigate the expression of constructed secretory αFGF gene in cardiomyocytes in vitro and in vivoand its angiogenic effect on host myocardium Methods: Secretory eucaryotic expression vector pCDNA3.1+SP αFGF was constructed and transfected in vitroto the neonate rat cardiomyocytes.The expression of αFGF was assessed.After transplantation of cells transfected with αFGF into the hearts of adult syngenic mice,the expression of αFGF in vivo and its angiogenic effect in host myocardium were studied. Results: The expression of αFGF was demonstrated by Western blotting and ELISA in the cultured media 24- 96 hours later. Using immunohistochemistry analysis, we detected αFGF in the recipient myocardium 96 hours later 20 days after cell transplantation, histological study revealed more blood vessels around the grafted cardiomyocytes in the host myocardium which was not marked in the controlled myocardium Conclusion: The plasmid we constructed can succeed in expressing and secreting αFGF peptidein vitroand in vivo,which would lead to angiogenesis in the recipient myocardiu

     

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