血小板表面N糖β1,6-GlcNAc分支缺陷诱导骨髓源性巨噬细胞表型M1转化

Defective N-glycanβ1,6-GlcNAc Platelets Promote M1 Phenotypic Switch in Polarization of Bone Marrow-Derived Macrophages

目的:探讨血小板N糖β1,6-GlcNAc分支缺陷对骨髓来源巨噬细胞(BMDMs)极化的影响。方法:通过苦马豆素灌胃动物模型获取N糖β1,6-GlcNAc分支缺陷血小板,流式细胞术检测血小板表面N糖β1,6-GlcNAc分支表达水平。将正常血小板和N糖支链缺陷血小板分别与BMDMs共同培养,采用流式细胞术、RT-PCR、ELISA等方法检测BMDMs的表型特征。结果:苦马豆素处理后的小鼠血小板N糖β1,6-GlcNAc表达较正常血小板明显减低(P<0.05)。与BMDMs共培养后,和正常血小板相比,N糖β1,6-GlcNAc分支缺陷血小板能够诱导巨噬细胞表达更多的诱导型一氧化氮合酶(iNOS)、CD16/32以及肿瘤坏死因子-ɑ(TNF-α),并降低CD206、Dectin-1、精氨酸酶-1(Arg-1)、白介素-10(IL-10)的表达。结论:N糖β1,6-GlcNAc分支缺陷血小板可在体外诱导BMDMs发生M1型极化,这可能是原发免疫性血小板减少症中免疫紊乱的发生机制之一。

Objective:To investigate the effects of defective N-glycanβ1,6-GlcNAc branch on platelets during the differentiation of bone marrow-derived macrophages(BMDMs)in vivo.Methods:Platelets with defective N-glycanβ1,6-GlcNAc was obtained by swainsonine gavage,which was confirmed by flow cytometry.BMDMs were co-cultured with normal or abnormal platelets.Then the phenotype of BMDMs was determined by flow cytometry,RT-PCR and ELISA.Results:Platelets in swainsoine-treated mice expressed lower N-glycanβ1,6-GlcNAc branch compared with that in normal mice.After co-cultured with BMDMs,abnormal platelets produced increased inducible nitric oxide synthase(iNOS),CD16/CD32 and tumor necrosis factor(TNF-ɑ),while the expression of CD206,Dectin-1,Arg-1 and IL-10 were down-regulated,compared with normal control.Conclusion:N-glycanβ1,6-GlcNAc-defective platelets could induce the differentiation of BMDMs into proinflammatory M1 phenotype in vitro,which would be probably involved in pathogenesis of primary immune thrombocytopenia.