中国全科医学Issue(30)：3727-3731,5.DOI:10.3969/j.issn.1007-9572.2015.30.020
益心解毒方对 NOX2亚基和 NOX4亚基过表达及小干扰 RNA 引起的心肌细胞还原型辅酶Ⅱ氧化酶活性变化的机制研究
Mechanism of Yixin Jiedu Formula on the Overexpression of NOX2 and NOX4 and the Myocardial NADPH ⅡOxidase Activity Changes Induced by RNAi
摘要
Abstract
Objective To observe the influence of Yixin Jiedu formula on NOX 2 and NOX4 overexpression and H9C2 myocardial NADPH Ⅱ oxidase activity changes induced by RNAi and investigate its mechanism.Methods We selected 16 normal male Sprague-Dawley rats and constructed the overexpression plasmids of NOX 2 and NOX4 and RNAi plasmids.Transfection reagents were employed to conduct transient transfection of H 9C2 myocardial cells, flow cytometry was undertaken to examine transfection efficiency.At 24 hours after transfection , the subjects were divided into normal H 9C2 cell group, overexpression group, H9C2 negative -turning control plasmid group, RNAi plasmid group, high -dose Yixin Jiedu formula group, middle-dose Yixin Jiedu formula group and low -dose Yixin Jiedu formula group.All the groups were intervened with different doses of drug serum , and NADPH oxidase activity was detected 24 hours after drug administration.Results The groups in which overexpression plasmids of NOX 2 was transfected were significantly different in NADPH oxidase activity ( P<0.05); overexpression group , high-dose Yixin Jiedu formula group and low -dose Yixin Jiedu formula group were significantly different from normal H9C2 cell group in NADPH oxidase activity (P<0.05); H9C2 negative-turning control plasmid group, RNAi plasmid group, middle -dose Yixin Jiedu formula group were significantly different from normal H 9C2 cell group in NADPH oxidase activity (P<0.05); RNAi plasmid group, high-dose Yixin Jiedu formula group and low -dose Yixin Jiedu formula group were significantly different from H9C2 negative-turning control plasmid group in NADPH oxidase activity (P<0.05); high-dose Yixin Jiedu formula group and low -dose Yixin Jiedu formula group were significantly different from RNAi plasmid group in NADPH oxidase activity ( P<0.05 ) ; middle-dose Yixin Jiedu formula group and high -dose Yixin Jiedu formula group were significantly different in NADPH oxidase activity ( P <0.05 ); low -dose Yixin Jiedu formula group and middle-dose Yixin Jiedu formula group were significantly different in NADPH oxidase activity ( P<0.05) .The groups in which overexpression plasmids of NOX 4 was transfected were significantly different in NADPH oxidase activity ( P <0.05 );overexpression group , high-dose Yixin Jiedu formula group and low -dose Yixin Jiedu formula group were significantly different from normal H9C2 cell group in NADPH oxidase activity (P<0.05); H9C2 negative -turning control plasmid group, RNAi plasmid group , middle-dose Yixin Jiedu formula group and low -dose Yixin Jiedu formula group were significantly different from overexpression group in NADPH oxidase activity (P<0.05); RNAi plasmid group, high-dose Yixin Jiedu formula group and low-dose Yixin Jiedu formula group were significantly different from H 9C2 negative-turning group in NADPH oxidase activity (P<0.05); high-dose Yixin Jiedu formula group and low -dose Yixin Jiedu formula group were significantly different from RNAi plasmid group in NADPH oxidase activity ( P<0.05 ); middle-dose Yixin Jiedu formula group and high -dose Yixin Jiedu formula group were significantly different in NADPH oxidase activity ( P<0.05 ); low-dose Yixin Jiedu formula group and middle-dose Yixin Jiedu formula group were significantly different in NADPH oxidase activity ( P<0.05 ) .Conclusion The overexpression of NOX 2 and NOX4 can significantly improve NADPH oxidase activity , and RNAi expression can silence NOX2 and NOX4, thereby significantly reducing NADPH oxidase activity.Yixin Jiedu formula doesn′t directly influence NADPH oxidase activity.The study shows the important mechanism of Yixin Jiedu formula may be that it can intervene and inhibit NADPH oxidase activity of NOX2 and NOX4, reduce the ROS level of myocardial muscle , protect myocardial cell and improve cardiac function.关键词
益心解毒方/肌细胞, 心脏/基因表达/小干扰RNA/氧化还原酶类Key words
Yixin jiedu formula/Myocytes, cardiac/Gene expression/RNAi/Oxidoreductases分类
医药卫生引用本文复制引用
解华,麻春杰,郭淑贞,付帮泽,冯玄超,王伟..益心解毒方对 NOX2亚基和 NOX4亚基过表达及小干扰 RNA 引起的心肌细胞还原型辅酶Ⅱ氧化酶活性变化的机制研究[J].中国全科医学,2015,(30):3727-3731,5.基金项目
国家自然科学基金资助项目（81102839）；科技部重大新药创制（2012ZX09103－201－011）；北京中医药大学自主选题 （）
