生脉注射液对蛋白激酶C调控LPS诱导iNOS基因表达的影响  

The effect of shengmai injection on the expression of iNOS regulated by protein kinase C

作　　者：杜少辉[1] 张悦 周大桥[1] 黄洁 魏志军[1] 

机构地区：[1]广东深圳市中医院内科,518033 [2]深圳市人民医院

出　　处：《中华急诊医学杂志》2003年第1期21-23,共3页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金项目 (3 9970 914 );广东省中医药管理局项目 (9844 7);深圳市科技局项目 (2 0 0 0 0 40 87)

摘　　要：目的　探讨生脉注射液对蛋白激酶C调控LPS诱导单核 /巨噬细胞iNOS基因表达的影响。方法　用蛋白激酶C (proteinkinaseC ,PKC)活性测定法研究LPS对PKC的激活作用及Griess还原法测定NO的生成 ;用基因重组技术构建iNOS荧光素酶报告基因载体 ;用基因转染及报告基因检测等方法研究生脉注射液调控LPS刺激RAW2 6 4 7细胞对iNOS启动子活性的诱导作用及其与PKC的关系。结果　生脉注射液可负调节LPS刺激RAW2 6 4 7细胞引起的PKC磷酸化激活和膜移位 ,并可延长PKC抑制剂H 7下调NO作用时间 ;荧光素酶表示基因实验结果显示 ,生脉注射液可抑制LPS刺激诱导的iNOS启动子的转录活性 ,并可增强H 7和钙离子通道阻滞剂Verapamil作用。结论　生脉注射液抑制LPS刺激RAW2 6 4 7细胞所致的NO生成增加 。Objective To explore the effects of shengmai ingection on the expression of Nitricoxide Synthase(iNOS) regulated by protein tinase C.Methods PKC activity assay was used to study the activation of PKC.Modified Griess method was used to determine inhibitory effect of PKC on the induction of NO by LPS.Luciferace reporter gene system of iNOS was constructed using gene recombination technique.Gene transfection and reporter gene assay were used to study the production of NO and induction of iNOS promoter transactivity regulated by Shengmai injection in RAW264 7 cell on the stimulation of LPS and the effect of PKC.Results In RAW 264 7 cells after the stimulation of LPS,PKC was activated by phosphorylation and translocated to inner cell membrane.Both of them were inhibited by Shengmai injection The stimulation of LPS on RAW264 7 cells decreased NO production,and increased,H 7 and Verapamil(a calcium ion channel blocker) showed an inhibitory effect on iNOS promoter transactivity induced by LPS.All effects were strengthened by Shengmai injection. Conclusion In RAW264 7cells stimulated by LPS,Shengmai injection can inhibit activation of PKC and the induction of iNOS gene expression,which contribute to the production of NO. [

关 键 词：生脉注射液 内毒素 诱导性一氧化氮合酶 基因表达 

分 类 号：R285.5[医药卫生—中药学]