益气解毒方水提物对RAW264.7细胞免疫功能的影响

Immunomodulating Effects of Yiqijiedu Formula on RAW264. 7 Cells

目的研究益气解毒方水提物对小鼠巨噬细胞系RAW264.7细胞免疫功能的影响,并探讨其可能的作用机制。方法用不同浓度益气解毒方水提物处理RAW264.7细胞,MTT法检测细胞增殖活性,ELISA法检测细胞肿瘤坏死因子-α(TNF-α)、白介素-12(IL-12)、一氧化氮(NO)的分泌水平,Western blot法检测细胞核转录因子NF-κBp65蛋白的表达水平。结果与空白对照组比较,益气解毒方水提物在10~320mg·L^(-1)范围内明显促进RAW264.7细胞增殖,且10 mg·L^(-1)浓度效果最佳(P<0.05);与空白对照组比较,益气解毒方水提物能促进TNF-α、IL-12及NO的分泌(P<0.05),显著诱导细胞内NF-κBp65蛋白的表达(P<0.05)。结论益气解毒方水提物对RAW264.7细胞的免疫功能具有调节作用,其作用可能与激活NF-κBp65蛋白通路相关。

Objective To investigate the immunomodulating effects of yiqijiedu formula on RAW264.7 cells and its possible mechanisms. Methods RAW264.7 ceils were treated with different concentrations of yiqijiedu formula,cell proliferation was measured by MTT assay; concentration of tumor necrosis factor-α（TNF-α） ,interleukin-12 （IL-12） and nitric oxide（NO） were determined by ELISA method; production of NF-κBp65 protein was detected by Western blot. Results Yiqijiedu formula（ 10-320 mg·L^-1 ）possessed the stimulation effect on macrophages proliferation in a dose-dependent manner（ P 〈 0.05 ） ; the production of TNF-α, IL-12 and NO were increased in a dose- dependent manner when pretreated with yiqijiedu formula（P 〈 0.05） ; the expression level of NF-κBp65 was significantly increased after yiqijiedu formula treatment （P 〈 0.05 ）. Conclusion Yiqijiedu formula could enhance the immune regulation of RAW264.7 cells,which may be related to the activation of NF-κBp65 pathway.