Physagulin H通过阻断NF-κB和JNK/MAPKs信号通路抑制LPS诱导RAW 264.7细胞产生的炎症反应

Physagulin H Inhibits LPS-Induced Inflammation in RAW 264.7 MacrophagesThrough Blocking NF-κB and JNK/MAPKs Signaling Pathways

采用脂多糖诱导的RAW 264.7巨噬细胞体外炎症模型评价physagulin H的抗炎活性.以MTT法评价细胞毒性,Griess法检测NO的含量,ELISA法检测PGE 2和TNF-α水平,Western Blot法检测炎症蛋白表达(iNOS和COX-2)以及对NF-κB炎性信号通路(IκB-α蛋白降解)和对MAPKs通路(JNK,p38和ERK蛋白磷酸化)的调控作用,研究physagulin H的抗炎活性及作用机制.结果表明,physagulin H可显著降低巨噬细胞RAW 264.7中NO、PGE 2和TNF-α的释放,还可显著降低iNOS和COX-2蛋白的高表达且呈浓度依赖性.PhysagulinH对LPS诱导NF-κB/IκB-α降解具有显著抑制作用.进一步的抗炎机制研究表明,physagulin H能抑制JNK磷酸化,但对ERK 1/2和p38磷酸化无明显抑制作用.综上,physagulin H是通过调控NF-κB和JNK/MAPKs信号转导通路下调LPS诱导的炎性蛋白的高表达,进而抑制小鼠巨噬细胞产生和释放过量炎症介质以及炎症因子,从而发挥抗炎作用.

The anti-inflammatory activity of physagulin H is evaluated by lipopolysaccharide(LPS)-induced RAW264.7 macrophage in vitro inflammation model.Cytotoxicity is evaluated by MTT assay,NO content is detected by Griess method,PGE 2 and TNF-αlevels are detected by ELISA,Western blot is used for detection of inflammatory protein expression(iNOS and COX-2)and relative proteins in NF-κB inflammatory signaling pathway(degradation of IκB-αprotein)and MAPKs pathway(phosphorylation of JNK,p38 and ERK proteins).The results show that physagulin H significantly reduces the release of NO,PGE 2 and TNF-αfrom macrophages RAW 264.7.In addition,physagulin H significantly reduces the overexpression of iNOS and COX-2 proteins.Physagulin H significantly inhibits the degradation of NF-κB/IκB-αinduced by LPS.Further anti-inflammatory mechanism studies show that physagulin H inhibits JNK phosphorylation but does not inhibit ERK 1/2 and p38 phosphorylation.In summary,physagulin H exerts anti-inflammatory effect by regulating the NF-κB signaling pathway to down-regulate the high expression of inflammatory proteins and then inhibit the release of the excessive inflammatory media and inflammatory cytokines induced by LPS.