不同来源白细胞微粒诱导巨噬细胞极化的比较研究

Comparative Study on Macrophage Polarization Induced by Different Sources of Leukocyte-derived Microparticles

目的探讨不同来源的白细胞微粒对于巨噬细胞亚型M1和M2的影响。方法诱导人单核细胞系(THP-1)细胞为巨噬细胞及泡沫细胞,提取巨噬细胞微粒(M-MPs)与泡沫细胞微粒(FC-MPs),将两种微粒分别加入巨噬细胞中。通过流式细胞仪观察微粒的大小;利用激光共聚焦显微镜观察微粒与巨噬细胞的结合状态;最后通过流式细胞仪检测M-MPs与FC-MPs分别诱导巨噬细胞后M1和M2亚型百分比。结果在细胞数相同的条件下,泡沫细胞产生的微粒数明显增多,大约是巨噬细胞产生微粒数的2倍(P<0.05)。镜下可见,两种微粒均与巨噬细胞接触或进入巨噬细胞内。与对照组相比,M-MPs与FC-MPs均可导致巨噬细胞向M1亚型极化;高浓度FC-MPs可显著抑制巨噬细胞向M2型极化,且M1/M2比值明显高于对照组,而M-MPs有降低M2亚型和M1/M2比值的趋势,但差异无统计学意义(P>0.05)。结论两种微粒均能与巨噬细胞结合,将巨噬细胞向促炎性转化,进一步促进炎症反应。相同条件下,泡沫细胞能够比巨噬细胞产生更多的FC-MPs,后者明显抑制巨噬细胞向抗炎型极化,加重促炎/抗炎的失衡。

Objective To explore the effects of leukocyte-derived microparticles from different sources on macrophage phenotypes M1 and M2.Methods THP-1 cells were induced to be macrophages and foam cells,then macrophage-derived microparticles(M-MPs)and foam cell-derived microparticles(FC-MPs)were extracted and used to induce macrophage.The size of MPs was tested by flow cytometry,and the binding state of MPs and macrophages was observed by laser confocal microscopy.Finally,the percentages of M1 and M2 phenotypes induced by M-MPs and FC-MPs were detected.Results Under the same number of cells,foam cells produced twice MPs compared with macrophages did(P<0.05).Both M-MPs and FC-MPs could bind to or enter macrophages observed by laser confocal microscopy.Compared with the control group,both M-MPs and FC-MPs could lead to macrophage polarization towards the M1(P<0.05).High concentration of FC-MPs inhibited macrophage polarization towards M2,and the M1/M2 ratio was higher than that in the control group significantly.Although M-MPs had a decreasing trend for M2 and M1/M2 ratio,there is no statistical difference(P>0.05).Conclusion Two kinds of microparticles can bind to macrophages and transform them into pro-inflammatory phenotypes,and further promoting the inflammatory response.Under the same conditions,foam cells produce more FC-MPs,which restrain macrophages from the anti-inflammatory phenotypes of macrophages,and exacerbate the pro-inflammatory/anti-inflammatory imbalance.