不同型号大孔树脂纯化的刘寄奴总黄酮对诱导型一氧化氮合酶的抑制作用  

iNOS Inhibitory Effect of Total Flavones of Artemisia anomala S. Moore Purified by Different Kinds of Macroporous Resin

作　　者：章丹丹[1] Jessica T.Li 章李军[1] 高月红[1] 潘一峰 李亚娟[1] 卞卡[1] 

机构地区：[1]上海中医药大学穆拉德中药现代化研究中心,上海201203 [2]上海现代中医药技术发展有限公司,上海200051

出　　处：《上海中医药杂志》2007年第5期70-73,共4页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家教育部重点科研基金资助项目(05ZZ11);上海市教委高校一氧化氮与炎症医学E-研究院计划项目(E-04010);上海市教委基金资助项目(05Jg05053);上海市科委基础研究重点基金资助项目(05JC14056)

摘　　要：目的考察不同型号大孔树脂纯化的刘寄奴总黄酮对IFN-γ和LPS协同诱导RAW264.7细胞中NO表达的影响。方法应用溶剂萃取和AB-8、D101、ADS-17大孔树脂分别制备刘寄奴总黄酮,分光光度法测定总黄酮的含量;IFN-γ和LPS协同诱导RAW264.7造成炎症模型;Griessreaction测定总黄酮对细胞上清液中亚硝酸盐含量的影响;RT-PCR考察总黄酮对细胞内诱导型一氧化氮合酶(iNOS)mRNA的影响;Westernblot考察总黄酮对细胞iNOS蛋白表达的影响。结果3种刘寄奴总黄酮含量均达到50%以上,且以ADS-17型大孔树脂富集所得的总黄酮含量最高,达到67.78%。3种刘寄奴总黄酮均呈剂量和时间依赖性抑制细胞上清液中亚硝酸盐的含量,同等流程中以ADS-17型大孔树脂富集的总黄酮达到半抑制率的剂量为最小。ADS-17型刘寄奴总黄酮呈剂量依赖性抑制iNOSmRNA和蛋白的表达。结论ADS-17型大孔树脂有效富集刘寄奴黄酮类化合物,适用于刘寄奴总黄酮的提取。ADS-17型刘寄奴总黄酮呈时间和剂量依赖性抑制细胞上清液中亚硝酸盐含量,部分通过抑制iNOSmRNA和蛋白的表达达到下调NO产量的结果。刘寄奴总黄酮可能是刘寄奴抗炎药效的物质基础,有望进一步开发成为iNOS抑制剂。Objective To evaluate the iNOS inhibitory effect of total flavones of Artemisia anomala S. Moore that was purified with three kinds of macroporous resin. Methods Total flavones were separated and purified from Artemisia anomala S. Moore by AB-8,D101,ADS-17 macroporous resin. Spectrophotometer was applied to quantify the total flavones. IFN-γ plus LPS stimulated expression of inducible nitric oxide synthase （iNOS） in RAW 264.7 macrophages has been used as inflammatory experimental model. Inhibition of iNOS by the total flavones were examined by RT-PCR for mRNA expression; Western blot for protein expression and Griess reaction for nitric oxide production. Results The purity of 3 total flavones extracted by AB-8,D101,ADS-17 respectively all reached required standard of 〉50%. The total flavones by ADS-17 was as high as 67.78%. All 3 total flavones from Artemisia anomala S. Moore exerted significant inhibition on IFN-γ plus LPS stimulated iNOS expression in RAW 264.7 macrophages which included mRNA and protein expressions as well as NO production by the enzyme. Conclusion ADS-17 macroporous resin showed highest capability to purify the total flavones from Artemisia anomala S. Moore. The total flavones purified all decreased iNOS mRNA and protein expressions in RAW264.7 cell in a dose and timedependent manner. The total flavones from Artemisia anomala S. Moore is a potential agent to develop as iNOS inhibitor.

关 键 词：刘寄奴总黄酮 炎症 一氧化氮 诱导型一氧化氮合酶 中药源抑制剂 

分 类 号：R285.5[医药卫生—中药学] R364.5[医药卫生—中医学]