中国全科医学2016，Vol.19Issue(15)：1774-1780,7.DOI:10.3969/j.issn.1007-9572.2016.15.008
建立β3肾上腺素能受体过表达的心肌成纤维细胞模型方法的比较
Comparison of Two Methods of Building Cardiac Fibroblasts Models Withβ3-AR Overexpression
摘要
Abstract
Objective To build neonatal SD rat cardiac fibroblasts(CFBs)models with β3-AR overexpression by two different methods and compare the two methods so as to provide new experimental method for exploring the effect of β3-AR on heart function,especially on chronic cardiac failure. Methods From March to June in 2015,8 SPF neonatal SD rats born 1 to 3 days ago were selected. CFBs of the rats were cultured in vitro and were randomly divided into three MOI groups which were MOI 50 group,MOI 100 group and MOI 200 group. These CFBs were infected by lentiviral vector with β3-AR overexpression,and infection rate was detected by flow cytometry to determine the optimal MOI. CFBs were randomly divided into four β3-AR agonist intervention groups which were normal control group,10 - 6 mol/ L group,10 - 5 mol/ L group,and 10 - 4 mol/ L group. Intervention was conducted on these CFBs by different concentrations of β3-AR agonist,and β3-AR expression level was detected by Western blotting method to find out the optimal concentration of β3-AR agonist in the intervention on CFBs. Results Observed under normal optics microscope,CFBs showed no pulsation and mostly took on fusiform shape,with large cell nucleus and transparent cytoplasm. The status of CFBs and fluorescent expression were observed under inverted fluorescence microscope at 24,48,72 and 96 h after infection. At 72 h,CFBs of each MOI group were in good growth status,they were mostly in fusiform shape and had no pulsation,with large cell nucleus,and the fluorescent expression observed by naked eye was optimal. MOI 100 group and MOI 200 group were higher than MOI 50 group in infection rate(P < 0. 05);MOI 200 group was lower than MOI 100 group in infection rate(P < 0. 05). 10 - 6 mol/ L group,10 - 5 mol/ L group and 10 - 4 mol/ L group were higher than normal control group in the expression level of β3-AR(P < 0. 05);10 - 5 mol/ L group was higher than 10 - 6 mol/ L group and 10 - 4 mol/ L group in the expression level of β3-AR(P < 0. 05). 10 - 5 mol/ L group and MOI 100 group were higher than normal control group in the expression level of β3-AR(P < 0. 05);MOI 100 group was higher than 10 - 5 mol/ L group in the expression level of β3-AR(P< 0. 05). Conclusion Two methods of lentiviral vector and β3-AR agonists are both able to induce the overexpression of β3-AR in CFBs. By comparison,the method of lentiviral vector infection is better.关键词
受体,肾上腺素能β3/成纤维细胞/心肌/慢病毒载体Key words
Receptors,adrenergic,beta-3/Fibroblasts/Myocardium/Lentiviral vecto分类
医药卫生引用本文复制引用
张源,王丽,张慧,马依彤,杨毅宁,马苗苗,朱晓丽..建立β3肾上腺素能受体过表达的心肌成纤维细胞模型方法的比较[J].中国全科医学,2016,19(15):1774-1780,7.基金项目
国家自然科学基金资助项目（81260028）；石河子大学科学技术研究发展计划“自然科学与技术创新”团队创新项目 （）
