摘要
目的 :研究非促分裂型haFGF(non mitogenichumanacidicfibroblastgrowthfactor,nm haFGF)真核表达载体 (pSecTag/nm haFGF)在原代培养的新生SD乳鼠心肌细胞中的分泌表达及其对心肌细胞生理特性的影响。方法 :采用脂质体转染的方法将真核表达载体pSecTag/nm haFGF转染原代培养的心肌细胞 ,转染 4 8h后用免疫印迹法 (westernblotting)检测nm haFGF的表达状况 ,同时用H2 O2 造成心肌细胞损伤 ,观察nm haFGF对心肌细胞的保护作用 ,并观察心肌细胞生理特性的变化。结果 :转染后 4 8h ,心肌细胞在转染nm haFGF基因后能够表达目标蛋白 ,而空载体实验组和未转染组均为阴性 ;MTT法检测结果表明 ,nm haFGF基因的表达产物具有生物活性 ,对H2 O2 造成的心肌细胞损伤有一定的保护作用 ;pSecTag/nm haFGF组的细胞搏动率明显高于阴性对照组 ,nm haFGF基因的表达产物对维持心肌细胞的生理特性具有积极的作用。结论 :转染nm haFGFcDNA能够在原代培养的SD乳鼠心肌细胞中分泌表达 ,并具有其相应的生理活性 ,对H2 O2
AIM:To investigate the expression effect of non mitogenic secretory human acidic fibroblast growth factor gene in primary cultured neonatal rat cardiomyocytes and its effect on the activity of these cells by building the eukaryotic expression plasmid which contains non motigentic secretory human acidic fibroblast growth factor gene (nm haFGF) METHOD:nm haFGF was transferred into the cultured neonatal SD rat ventricular myocytes by the way of liposome mediated transfection The transfective effect of pSecTag/nm haFGF was tested by western blotting;Meanwhile,we measured the antioxidity of the transfection nm haFGF cDNA groups and the control groups by MTT method,and observed the change of cardiomyocytes physiological character RESULT: After 48 h of transfection,the results of western blotting were positive in the liposome/pSecTag nm haFGF group,but the other control groups were negative Result of MTT indicated that the transfective nm haFGF group had significantly higher cell viability than the control group The spontaneous beat rate transfective nm haFGF group was significantly higher than the other control groups CONCLUSION:The eukaryotic plasmid can succeed in expressing and secreting functional protein in the primacy cultured neonatal rat ventricular myocytes,and has corresponding bioactivity on the cardiomyocytes,and show the corresponding cardioprotective effect on H 2 O 2
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2004年第2期168-172,共5页
Journal of China Pharmaceutical University
基金
国家 8 63计划资助项目 (No.2 0 0 1AA2 15 13 1)
国家"十五"重大专项资助项目 (No.2 0 0 2AA2Z3 3 18)~~
关键词
酸性成纤维细胞生长因子
脂质体介导
转染
心肌细胞
Non motigentic secretory human acidic fibroblast growth factor
Liposome mediated
Transfection
Cardiomyocytes
