黄芪甲苷调控STAT1/IκB/NF-κB信号通路抑制γ-干扰素诱导BV-2细胞激活  

Astragaloside Ⅳ regulates STAT1 / IκB / NF-κB signaling pathway to inhibit activation of BV-2 cells

作　　者：贺一新[1,2] 石海莲[2] 刘宏帅[2] 吴辉[2] 张蓓蓓[2] 吴晓俊[2] 王峥涛[1,2] 

机构地区：[1]中国药科大学中药学院,江苏南京210009 [2]上海中医药大学中药研究所暨上海市复方中药重点实验室,上海201203

出　　处：《中国中药杂志》2015年第1期124-128,共5页China Journal of Chinese Materia Medica

基　　金：教育部高等学校博士学科点专项科研基金优先发展领域项目(20123107130002);上海市教委科研创新重点项目(13ZZ099);上海高校特聘教授(东方学者)岗位计划项目(2013-59)

摘　　要：目的:研究黄芪甲苷(ASI)对γ-干扰素(IFN-γ)诱导小胶质细胞激活的抑制作用及机制。方法:不同浓度的ASI(25,50,100μmol·L-1)预处理BV-2小胶质细胞2 h后,以IFN-γ刺激1.5 h或24 h,分别收集细胞培养基上清和细胞。分别以Griess和ELISA法检测培养基中一氧化氮(NO)和肿瘤坏死因子α(TNF-α)含量;q PCR法检测细胞中CD11b,TNF-α,白介素1β(IL-1β)和诱导型一氧化氮合酶(i NOS)mRNA水平;Western blot法检测细胞STAT1/IκB/NF-κB信号通路蛋白表达。结果:ASI能显著抑制IFN-γ诱导BV-2细胞NO和TNF-α水平的升高(P<0.001);进一步研究表明,50μmol·L-1ASI能够显著抑制细胞IL-1β和TNF-α的mRNA表达(P<0.01,P<0.05),并表现出下调i NOS mRNA表达的趋势,但对BV-2细胞CD11b的mRNA水平无显著影响;同时,ASI显著抑制IFN-γ诱导上调的p STAT1,p IκB和p NF-κB的蛋白表达。结论:ASI能够抑制IFN-γ诱导的小胶质细胞的激活,其机制与其抑制STAT1/IκB/NF-κB信号通路的激活,降低炎症状态下小胶质细胞IL-1β,TNF-α以及i NOS的基因表达,从而减少NO和TNF-α的生成有关。Objective： The study was aimed to investigate the inhibitory effect and mechanism of astragaloside IV （ASI） on the activation of microglial cells. Method ： After pre-incubated with ASI for 2 h, microglial cells BV-2 were stimulated with interferon-γ （IFN-γ） for 1.5 h and 24 h, respectively. Secretion of nitric oxide （NO） in the medium was measured by Griess method. Production of tumor necrosis factor alpha （TNF-α） was detected by ELISA approach. Cellular gene expressions of CD1 l b, TNF-α, interleukin 1β （IL-1β） and induced nitric oxide synthase （iNOS） were examined by quantitative -PCR analysis. Total and phosphorylation of STAT1, IKB and NF-KB was analyzed by Western blot method. Result： ASI could significantly inhibit the increased secretion of TNF-α and NO from BV-2 cells upon IFN-T stimulation （P 〈0. 001 ）. Further study showed that ASI significantly down-regulated gene expression of IL-1β and TNF-α （P 〈0. 01, P 〈0.05） and exhibited a trend to reduce that of iNOS. IFN-γ and ASI have no obvious effect on gene expression of CDllb. Moreover, ASI inhibited the phosphorylation of STAT1, IKB and NF-kB elicited by IFN-γ stimulation. Conclusion： ASI could restrain microglial activation through interfering STAT1/IkB/NF-kB signaling pathway, reducing gene expression of IL-1β and TNF-α, and thus inhibiting the production of proinflammatory mediators such as NO and TNF-α.

关 键 词：黄芪甲苷 小胶质细胞 神经炎症 STAT1 IΚB NF-κB 

分 类 号：R285[医药卫生—中药学]