人单核细胞和外周血单个核细胞衍生的巨噬细胞极化特性的比较

Comparison of polarization characteristics of human monocyte cell- and peripheral blood mononuclear cell-derived macrophages

目的在成功建立巨噬细胞(Mφ)体外培养模型基础上,比较不同来源Mφ的极化特性。方法采用密度梯度离心法和免疫磁珠法体外分离并培养外周血单个核细胞(PBMC),经人重组巨噬细胞集落刺激因子诱导后培养出PBMC Mφ。人单核细胞系THP-1在100 nmol·L^-1佛波酯刺激下分化为THP-1 Mφ。上述两种来源的Mφ分别采用不同的极化因子进行诱导,经50 ng·mL^-1脂多糖+20 ng·mL^-1干扰素-γ诱导形成M1型Mφ(M1 Mφ),经20 ng·mL^-1白细胞介素-4诱导形成M2型Mφ(M2 Mφ)。观察两种来源Mφ的细胞形态,比较两种细胞表面极化标记物(CD86和CD206)和细胞因子[白细胞介素-1β(IL^-1β)、肿瘤坏死因子-α(TNF-α)、甘露糖受体C1样蛋白(MRC1)、白细胞介素-10(IL^-10)等]的表达情况。结果两种来源的Mφ经M1极化因子刺激后,TNF-α、IL^-1β的mRNA水平显著上调,CD86表达量增加,与PBMC Mφ相比,THP-1 Mφ中的TNF-α、IL^-1β分泌水平更高;两种Mφ在M2极化因子刺激后MRC1、IL^-10 mRNA水平上调,CD206在PBMC Mφ中表达显著增强,而在THP-1 Mφ中表达没有明显变化。结论THP-1可能更适合进行M1极化的研究,而PBMC可能更适合进行M2极化的研究。

Objective Obtained from successfully established polarization models,the polarization characteristics of macrophages(Mφ)from different sources were compared to provide an experimental basis for biological treatments.Methods Peripheral blood mononuclear cells(PBMC)were isolated through density gradient centrifugation and magnetic cell sorting and were cultured in vitro.PBMC Mφwas induced by human recombinant macrophage colony-stimulating factor.The human monocyte cell line THP-1 was differentiated into THP-1 Mφunder the induction of 100 nmol·L^-1 phorbol 12-myristate 13-acetate.Morphology,polarization marker expression(CD86 and CD206),and cytokine secretion levels of the two groups were compared under the condition of M1 or M2 polarization factors.M1 polarization factors was 50 ng·mL^-1 lipopolysaccharides and 20 ng·mL^-1 interferon-γ,and M2 polarization factors was 20 ng·mL^-1 interleukin-4.Results After M1 polarization,the mRNA levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL^-1β)were markedly upregulated,the expression of CD86 was enhanced,and the secretions of IL^-1βand TNF-αin THP-1 Mφwere higher than those in PBMC Mφ.After M2 polarization,the mRNA levels of mannose receptor C-type 1 and interleukin-10 increased,and the expression of CD206 was upregulated in PBMC Mφ.However,no difference was observed in THP Mφ.Conclusion THP-1 is appropriate for M1 polarization,and PBMC is appropriate for M2 polarization.