日本血吸虫高迁移率族蛋白活化鼠巨噬细胞的初步研究

摘要

为研究日本血吸虫高迁移率族蛋白(Schistosoma japonicum high mobility group box1,Sj HMGBl)体外活化小鼠巨噬细胞的功能,利用真核重组SjHMGB1蛋白与巨噬细胞共培养48 h,流式细胞术检测巨噬细胞表面标志分子以及趋化因子受体的表达.收集Sj HMGB1蛋白与巨噬细胞共孵育48 h后上清,ELISA检测上清中TNF-α与IL-10的含量,Griess法检测上清中NO的含量.流式结果显示,与对照组相比,Sj HMGB1蛋白刺激组的巨噬细胞表面MHC-Ⅱ分子,TLR4受体以及趋化因子受体CCR7的表达显著上调且差异具有极显著统计学意义(P＜0.01),TLR2受体的表达显著下调,差异具有显著统计学意义(P.＜0.05).ELISA结果显示,Sj HMGB1能够刺激巨噬细胞分泌致炎因子TNF-α,未能促进抑炎因子IL-10的释放.Griess法表明Sj HMGB1能够促进巨噬细胞分泌NO.本研究结果表明研HMGB1可能通过TLR4通路活化小鼠巨噬细胞并诱导其向致炎性M1型巨噬细胞极化.%This study aimed at determining the activation of high mobility group boxl protein (SjHMGB 1)of Schistosomajaponicum on mouse macrophagocyte.When Recombinant SfHMGB 1 protein from eukaryotic cell expression system was incubated with macrophage for 48 h,the expression of macrophage surface molecules and chemokine receptors were detected by flow cytometry.TNF-α and IL-10 in the culture supernatant were detected by ELISA,and the content of NO in supernatant was assayed by Griess method.Results showed that,compared with the control group,the expression levels of MHC-11 molecules,TLR4 receptor and chemokine receptor CCR7 on the surface of macrophage stimulated by SjHMGB1 group were significantly increased (P ＜ 0.01),while TLR2 receptor was signi ficantly decreased (P＜0.05).The results of test by ELISA demonstrated that SjHMGB 1 could stimulate the secretion of inflammatory factor TNF-α,and fail to promote the release of IL-10 on macrophage.The result of test using Griess method showed that SjHMGB 1 could promote the secretion of NO on macrophage.The data suggest that SjHMGB1 could activate mouse macrophages via TLR4 and induce macrophages to undergo M l-type inflammatory polarization.