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(1)
[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(25，50，100 μmol·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;qPCR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(iNOS)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，50 μmol·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iNOS 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 pSTAT1，pIκB和pNF-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iNOS的基因表达，从而减少NO和TNF-α的生成有关。
, http://www.100md.com
[关键词]黄芪甲苷;小胶质细胞;神经炎症;STAT1;IκB;NF-κB
[收稿日期]2014-07-15
[基金项目]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)
[通信作者]*吴晓俊，研究员，Tel：(021)51322578，E-mail：xiaojunwu320@126.com;*王峥涛，教授，Tel：(021)51322507，E-mail：ztwang@shutcm.edu.cn
[作者简介]贺一新，博士研究生，E-mail：29693613@qq.com
Astragaloside IV regulates STAT1/IκB/NF-κB signaling pathway
, 百拇医药
to inhibit activation of BV-2 cells
HE Yi-xin1，2， SHI Hai-lian2， LIU Hong-shuai2， WU Hui2， ZHANG Bei-bei2， WU Xiao-jun2*， WANG Zheng-tao1，2*
(1. Department of Pharmacognosy， School of Traditional Chinese Medicine， China Pharmaceutical University， Nanjing 210009， China;
2. Shanghai Key Laboratory of Complex Prescriptions， Institute of Chinese Materia Medica， Shanghai University of
, http://www.100md.com
Traditional Chinese Medicine， Shanghai 201203， China)
[Abstract]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 CD11b， TNF-α， interleukin 1β (IL-1β) and induced nitric oxide synthase (iNOS) were examined by quantitative -PCR analysis. Total and phosphorylation of STAT1， IκB and NF-κB 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-γ 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 CD11b. Moreover， ASI inhibited the phosphorylation of STAT1， IκB and NF-κB elicited by IFN-γ stimulation. Conclusion： ASI could restrain microglial activation through interfering STAT1/IκB/NF-κB 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., 百拇医药(贺一新 石海莲 刘宏帅 吴辉 张蓓蓓 吴晓俊 王峥涛)
, http://www.100md.com
[关键词]黄芪甲苷;小胶质细胞;神经炎症;STAT1;IκB;NF-κB
[收稿日期]2014-07-15
[基金项目]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)
[通信作者]*吴晓俊，研究员，Tel：(021)51322578，E-mail：xiaojunwu320@126.com;*王峥涛，教授，Tel：(021)51322507，E-mail：ztwang@shutcm.edu.cn
[作者简介]贺一新，博士研究生，E-mail：29693613@qq.com
Astragaloside IV regulates STAT1/IκB/NF-κB signaling pathway
, 百拇医药
to inhibit activation of BV-2 cells
HE Yi-xin1，2， SHI Hai-lian2， LIU Hong-shuai2， WU Hui2， ZHANG Bei-bei2， WU Xiao-jun2*， WANG Zheng-tao1，2*
(1. Department of Pharmacognosy， School of Traditional Chinese Medicine， China Pharmaceutical University， Nanjing 210009， China;
2. Shanghai Key Laboratory of Complex Prescriptions， Institute of Chinese Materia Medica， Shanghai University of
, http://www.100md.com
Traditional Chinese Medicine， Shanghai 201203， China)
[Abstract]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 CD11b， TNF-α， interleukin 1β (IL-1β) and induced nitric oxide synthase (iNOS) were examined by quantitative -PCR analysis. Total and phosphorylation of STAT1， IκB and NF-κB 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-γ 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 CD11b. Moreover， ASI inhibited the phosphorylation of STAT1， IκB and NF-κB elicited by IFN-γ stimulation. Conclusion： ASI could restrain microglial activation through interfering STAT1/IκB/NF-κB 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., 百拇医药(贺一新 石海莲 刘宏帅 吴辉 张蓓蓓 吴晓俊 王峥涛)