【摘要】 通过对M1和M2型巨噬细胞表型相关指标的比较分析,评价各鉴定巨噬细胞类型的表型指标及其意义。按常规方法以IFN-γ及LPS将骨髓来源巨噬细胞诱导成M1型巨噬细胞,以IL-4诱导出M2型巨噬细胞。分别以RT-PCR和酶活性定量方法检测精氨酸代谢相关酶的表达和活性;以ELISA检测IL-12和IL-10的分泌;以FACS检测巨噬细胞膜分子的表达。结果显示:M1型巨噬细胞诱导性一氧化氮合酶(inducible nitric oxide synthase,iNOS)表达和活性水平较未刺激组明显升高,IL-12产生显著增加,CD16/32表达上调;而M2型巨噬细胞I型精氨酸酶(arginase 1,Arg-1)的表达水平和酶活性较未刺激巨噬细胞显著提高,IL-10分泌轻度增加,并且表达高水平的CD206和DECTIN-1。表型比较分析结果表明,iN-OS表达和活性、IL-12的分泌和膜蛋白CD16/32可用于鉴定M1型巨噬细胞,而Arg-1、CD206和DECTIN-1是鉴定M2型巨噬细胞较为理想的表型指标。更多还原

【Abstract】 To well identify and evaluate the phenotypic characteristics of M1 and M2 macrophages by quantitative comparative analysis,bone marrow derived macrophages were stimulated with murine IFN-γ plus LPS or IL-4 for induction of M1 and M2 macrophages respectively.The expression of inducible nitric oxide synthase(iNOS) and arginase 1(Arg-1)was measured by RT-PCR.The iNOS and Arg-1 activity was analyzed by Griess assay and arginase assay,respectively.IL-12 and IL-10 levels were quantified by ELISA assay.Surface markers were determined by FACS analysis.It showed that IFN-γ plus LPS remarkably induced NO release and iNOS mRNA expression in macrophages,which also produced higher abundances of IL-12 compared to untreated macrophages.CD16/32 expression was upregulated about 5-fold in the presence of IFN-γ and LPS.After stimulation with IL-4,macrophages showed high levels of Arg-1 expression and enzyme activity.FACS analysis revealed high expression of CD206 and DECTIN-1 on M2 macrophages.Comparative analysis indicated that phenotypes including iNOS,IL-12 and CD16/32 could be well used for identification of M1 macrophages,whereas Arg-1,CD206 and DECTIN-1 for identification of M2 macrophages.更多还原