【摘要】 目的研究低氧对培养大鼠心肌细胞蛋白激酶A(protein kinase A,PKA)与SCN5A基因表达的影响。方法将原代培养的大鼠乳鼠心肌细胞随机分为5组:对照组、低氧(5%CO2+2%O2+93%N2)培养1h组、6 h组、12 h组和24 h组,利用非放射性方法测定各组细胞内PKA活性,利用RT-PCR和Western blot等方法检测各组SCN5A基因表达情况。结果各组PKA活性:低氧1h组和低氧6 h组高于正常组(P<0.01),低氧12 h组和24 h组低于正常组(P<0.01)。SCN5AmRNA相对表达量与蛋白质表达量:低氧1h组和低氧6h组显著高于对照组;低氧12 h组和低氧24 h组显著低于对照组。结论低氧对培养细胞内PKA活性与SCN5A基因的表达有一定调节作用,且对PKA的作用早于对SCN5A基因表达的作用,PKA可能参与影响大鼠心肌细胞缺氧后SCN5A基因的表达。更多还原

【Abstract】 Objective To study the effect of hypoxia on the activity of protein kinase A(PKA) and the expression of SCN5A in cultured rat myocardial cells.Methods The cultured myocardial cells of rats were divided into 5 groups:cultured in normal condition group,cultured in hypoxic condition for 1 h,6 h,12 h and 24 h group.Activity of PKA were determined by PepTag Non-Radioactive cAMP-Dependent Protein Kinase Assay,SCN5A mRNA and protein were examined respectively by RT-PCR and Western blot at different time of hypoxic culture.Results The activity of PKA in endochylema and the expression of SCN5A at the level of mRNA and protein increased significantly on the points of 1 h and 6 h(P<0.01),but decreased on the 12 h and 24 h points.Conclusion Hypoxia has a role to the activity of PKA and the expression of SCN5A in cultured rat myocardial cells.PKA may affect the expression of SCN5A.更多还原