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目的 观察分泌型aFGF真核表达载体转染大鼠心肌细胞后 ,aFGF在体内、外的分泌表达及转染心肌细胞的移植对宿主心肌血管形成作用的影响。方法 构建分泌型aFGF真核表达载体(pCDNA3 1+SP aFGF) ,转染培养大鼠的心肌细胞 ,检测体外心肌细胞对aFGF的分泌表达 ;将转染细胞移植入同种大鼠心肌 ,检测移植细胞在体内对aFGF的表达及宿主心肌的血管生成。结果 硝酸纤维素膜点杂交和ELISA显示 ,aFGF在转染 2 4～ 96h细胞培养上清中有较高的表达 ;接受转染心肌细胞移植96h后免疫组化检查 ,发现aFGF在受体心肌内得到分泌表达 ,2 0d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aFGF真核表达载体的心肌细胞 ,在体内、外能分泌表达aFGF ;通过心肌细胞移植 ,分泌的aFGF在受体心肌组织中有较强的血管生成作用
Objective To observe the effect of aFGF in vitro and in vivo on the secretion of aFGF and the effect of transfection of cardiomyocytes on the formation of aortic myocardium after transfection of aFGF secreting eukaryotic expression vector into rat cardiomyocytes. Methods The eukaryotic expression vector of secreting aFGF (pCDNA3 1 + SP aFGF) was constructed and transfected into rat cardiomyocytes to detect the secretion of aFGF from cardiomyocytes in vitro. Transfected cells were transplanted into the same myocardium of rats, AFGF expression in the body and angiogenesis in the host myocardium. Results The results of nitrocellulose membrane dot blot and ELISA showed that aFGF was highly expressed in the supernatant of cells transfected for 24 to 96 hours. After transplanted for 96 hours, aFGF was detected in the myocardium Secreted expression was obtained. Twenty days later, tissue sections were found to have more new capillaries in the myocardium around the transplanted cardiac myocytes. Conclusions AFPs secreting aFGF eukaryotic expression vector can express aFGF both in vitro and in vivo. The aFGF secreted by cardiac myocyte transplantation has strong angiogenesis in the myocardium of recipient