黄芪总黄酮对RAW264.7细胞细胞因子和介质分泌水平的影响

Effects of total flavonoids of Astragalus on secretion levels of cytokines and mediators in RAW264.7 cells under normal conditions

试验采用传统中药黄芪的主要成分黄芪总黄酮为研究对象,探讨了黄芪总黄酮的体外免疫作用。以RAW264.7细胞为模型,通过细胞毒性试验拟定黄芪总黄酮的试验剂量,分别设立药物低、中、高剂量组以及空白组,通过ELISA法测定细胞因子IL-6、IL-1β、IFN-γ、TNF-α和介质PGE2的含量,Griess法测定介质NO分泌水平,Western blot法分析iNOS和COX-2蛋白含量变化。结果表明:黄芪总黄酮剂量依赖性的促进了正常条件下RAW264.7细胞上清液中细胞因子IL-6、IL-1β、IFN-γ、TNF-α和细胞介质NO、PGE2的分泌,上调了iNOS和COX-2蛋白的表达,表明黄芪总黄酮具有体外免疫增强作用。

In this study, we investigated the immunological effects of total flavonoids of astragalus （TFA） in vitro which is the main components of traditional Chinese medicine Astragalus. The experimental dose of TFA was established through cytotoxicity test by using the RAW264.7 cells. TFA groups （low, medium and high doses） and blank group were set up, respectively. The levels of cytokines IL-6, IL-1β, IFN-γ, TNF-α, mediator PGE2 were detected by ELISA kits, NO secretion level was detected by Griess reagent and COX-2, iNOS protein expression levels were determined by Western blot. The results showed that the levels of IL-6, IL-1β, IFN-γ, TNF-α, NO and PGE2 were significantly increased, and the expression levels of iNOS and COX-2 proteins were up-regulated in TFA treated groups by dose dependent. It is shown that TFA could enhance immunity in vitro .