去氢丹参新酮对神经炎症的抑制作用及机制研究  

The anti-neuroinflammatory effects of dehydromiltirone and related mechanisms

作　　者：李德川[1] 鲍秀琦[1] 张德武[1] 孙华[1] 戴均贵[1] 张丹[1] 

机构地区：[1]中国医学科学院北京协和医学院药物研究所天然药物活性物质与功能国家重点实验室,北京100050

出　　处：《中国药理学通报》2016年第2期177-183,共7页Chinese Pharmacological Bulletin

基　　金：国家重点基础研究发展计划(973计划)资助项目(No2011CB504105);教育部新世纪优秀人才(No3332013128);北京市科技新星项目(No 2011109);北京市优秀人才(No 2012D0009008000005)

摘　　要：目的研究去氢丹参新酮对脂多糖(lipopolysaccharide,LPS)诱导小胶质细胞系BV2细胞产生炎症反应的抑制作用及其作用机制。方法不同浓度去氢丹参新酮预孵育BV2细胞后,用LPS刺激引起神经炎症相关反应。Griess试剂法检测去氢丹参新酮对活化的BV2细胞产生一氧化氮(nitric oxide,NO)的影响,ELISA检测细胞上清液中TNF-α和IL-6的释放量,Confocal观察小胶质细胞表面活化标志物MAC-1表达量的变化,Western blot检测炎症相关信号通路蛋白表达的变化。结果去氢丹参新酮可明显抑制LPS刺激BV2细胞产生的炎症因子包括NO、TNF-α和IL-6的水平,同时抑制一氧化氮合酶、环氧合酶-2等炎症相关蛋白的表达和小胶质细胞表面活化标志物MAC-1的表达。机制研究发现,去氢丹参新酮对PI3K/Akt的过度磷酸化以及NF-κB的过度活化都有明显的抑制作用。结论去氢丹参新酮具有很好的抑制神经炎症活性,其作用机制可能是通过PI3K/Akt信号通路抑制NF-κB的活化而实现的。Aim To investigate the anti-neuroinflammatory activities of dehydromiltirone and the underlying mechanisms in LPS-stimulated microglial cell line BV2 cells.Methods BV2 cells were pre-treated with dehydromiltirone,then stimulated by LPS.The levels of nitric oxide(NO) were measured by Griess assay,and the concentrations of pro-inflammatory cytokines were measured by ELISA assay.Confocal fluorescence microscopy was used to measure the expression of MAC-1,the biomarker of activated BV2 cells.The levels of—inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),NF-κB and PI3K/Akt were determined by Western blot analysis.Results The treatment of dehydromiltirone significantly inhibited the production of NO,TNF-α and IL-6,attenuated the expression of iNOS and COX-2 protein,and dampened the microglial activation in LPS-stimulated BV2 cells.The mechanistic study revealed that dehydromiltirone inhibited the phosphorylation of PI3 K and Akt in LPSstimulated BV2 cells,and decreased NF-κB activation by suppressing the degradation of IκB.Conclusion dehydromiltirone shows significant anti-neuroinflammatory effects through inhibiting PI3K/Akt phosphorylation and then inhibiting NF-κB signaling pathway.

关 键 词：去氢丹参新酮 神经炎症 小胶质细胞 NFκB PI3K/AKT 脂多糖 

分 类 号：R284.1[医药卫生—中药学] R322.8[医药卫生—中医学]