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通过分析日本血吸虫高迁移率族蛋白B1(Schistosoma japonicum high mobility group protein B1,SjHMGB1)刺激的巨噬细胞的表型相关分子的表达情况,研究SjHMGB1诱导巨噬细胞向M1型极化的功能。分别用LPS及IFN-γ和IL-4诱导小鼠RAW264.7巨噬细胞成M1型和M2型作为阳性对照组,以SjHMGB1诱导的巨噬细胞为实验组。分别利用Griess法、FCM、RTPCR和酶活性检测巨噬细胞中iNOS、CD16/32、ArginaseⅠ、CD206等主要标志分子。结果与空白对照组相比,SjHMGB1刺激组和M1型阳性对照组巨噬细胞NO分泌显著增加,精氨酸酶活性变化不显著,M2型巨噬细胞呈现相反的表型。流式结果显示SjHMGB1刺激组和M1型阳性对照组的巨噬细胞诱导型一氧化氮合成酶(iNOS)和CD16/32表达上调,而CD206变化不明显。RT-PCR结果显示SjHMGB1蛋白刺激组和M1型阳性对照组的巨噬细胞iNOS表达上调,而arginaseⅠ和CD206变化不明显;M2型巨噬细胞arginaseⅠ表达水平明显升高,CD206表达上调。结果表明SjHMGB1能够诱导巨噬细胞往M1型巨噬细胞分化。
The function of SjHMGB1-induced macrophages to M1 polarization was investigated by analyzing the expression of phenotype-related molecules in macrophages stimulated with Schistosoma japonicum high mobility group protein B1 (SjHMGB1). Mouse RAW264.7 macrophages were induced into M1 and M2 by LPS, IFN-γ and IL-4 respectively as positive control group and macrophages induced by SjHMGB1 as experimental group. The major marker molecules such as iNOS, CD16 / 32, Arginase I and CD206 in macrophages were detected by Griess method, FCM, RTPCR and enzyme activity respectively. Results Compared with the blank control group, the NO secretion of macrophages in SjHMGB1-stimulated group and M1-positive control group was significantly increased, the arginase activity was not significantly changed, M2-type macrophages showed the opposite phenotype. The results of flow cytometry showed that the expressions of iNOS and CD16 / 32 were up-regulated in SjHMGB1 stimulation group and M1 positive control group, while the changes of CD206 were not obvious. The results of RT-PCR showed that the iNOS expression of macrophages in SjHMGB1 stimulation group and M1 positive control group was up-regulated, while the changes of arginase I and CD206 were not obvious. The expression of arginase I in M2 macrophage increased and the expression of CD206 was up-regulated. The results showed that SjHMGB1 can induce macrophages to differentiate into M1 macrophages.