摘要
目的 探讨白细胞介素(IL)-34对人外周血单核细胞来源的巨噬细胞极化和迁移能力的影响.方法 采用免疫磁珠分选法从人外周血单核细胞中分选CD14+单核细胞,并使用流式细胞仪检测CD14+单核细胞纯度.将CD14+单核细胞分为M1型组、IL-34-M1型组、M2型组、IL-34-M2型组,M1型组和IL-34-M1型组细胞加入粒细胞-巨噬细胞集落刺激因子(GM-CSF)诱导5 d后,半量换液,加入干扰素-γ、脂多糖、IL-6和GM-CSF继续诱导4 d;M2型组和IL-34-M2型组细胞加入巨噬细胞集落刺激因子(M-CSF)诱导5 d后,半量换液,加入M-CSF、IL-4、IL-6和IL-13继续诱导4 d.IL-34-M1型组和IL-34-M2型组细胞于诱导开始和诱导第5天加入IL-34进行共诱导.诱导第9天,采用流式细胞术检测各组细胞中CD14+CD86+细胞(M1型巨噬细胞)和CD14+CD163+细胞(M2型巨噬细胞)比例,采用Transwell小室实验检测M2型组、IL-34-M2型组细胞迁移能力.结果 通过磁珠分选法获得了高纯度的CD14+单核细胞,CD14阳性率为(96.77±2.72)％,可用于巨噬细胞的诱导.M1型组、IL-34-M1型组中CD14+CD86+细胞比例分别为(43.20±7.59)％、(27.87±2.06)％,M2 型组、IL-34-M2 型组中 CD14+CD163+细胞比例分别为(47.70± 4.49)％、(58.95±3.65)％;IL-34-M1 型组中 CD14+CD86+细胞比例显著低于 M1 型组(P＜0.05),IL-34-M2 型组中CD14+CD163+细胞比例显著高于M2型组(P＜0.05).M2型组、IL-34-M2型组巨噬细胞迁移数分别为(97.8±9.0)、(205.6±21.9)个;IL-34-M2型组巨噬细胞迁移数显著多于M2型组(P＜0.05).结论 IL-34可抑制人单核细胞来源的巨噬细胞向M1型极化,促进巨噬细胞向M2型极化,并可增强M2型巨噬细胞的迁移能力.
Abstract
Objective To investigate the effects of interleukin(IL)-34 on the polarization and migration ability of macrophages derived from human peripheral blood monocytes.Methods The CD14+monocytes were isolated from human peripheral blood monocytes by immunomagnetic bead sorting,and the purity of CD14+monocytes was detected by flow cytometry.The CD14+monocytes were divided into M1 type group,IL-34-M1 type group,M2 type group and IL-34-M2 type group.The cells in the M1 type group and IL-34-M1 type group were added granulocyte-macrophage colony stimulating factor(GM-CSF)to induce for 5 days,and half of the fluid was changed,and then the interferon-γ,lipopolysaccharides,IL-6 and GM-CSF were added for another 4-day induction;the cells in the M2 type group and IL-34-M2 type group were added macrophage colony stimulating factor(M-CSF)to induce for 5 days,and half of the fluid was changed,and M-CSF,IL-4,IL-6,and IL-13 were added for another 4-day induction.The cells in IL-34-M1 group and IL-34-M2 group were co-induced with IL-34 at the beginning of induction and on the 5th day of induction.On the 9th day of induction,the proportion of CD14+CD86+cells(M1 type macrophages)and CD14+CD163+cells(M2 type macrophages)in each group was detected by flow cytometry,and the migration ability of cells in the M2 type group and IL-34-M2 type group was detected by the Transwell chamber experiments.Results High purity CD14+monocytes were obtained through magnetic bead sorting,with a CD14 positive rate of(96.77± 2.72)％,which could be used for macrophage induction.The proportion of CD14+CD86+cells in the M1 type group and IL-34-M1 type group was(43.20±7.59)％and(27.87±2.06)％,respectively.The proportion of CD14+CD163+cells in the M2 type group and IL-34-M2 type group was(47.70±4.49)％and(58.95±3.65)％,respectively;the proportion of CD14+CD86+cells in the IL-34-M1 type group was significantly lower than that in the M1 type group(P＜0.05),while the proportion of CD14+CD163+cells in the IL-34-M2 type group was significantly higher than that in the M2 type group(P＜0.05).The number of migrating cells of macrophages in the M2 type group and IL-34-M2 type group was 97.8±9.0 and 205.6±21.9,respectively;the number of migrating cells of macrophages in the IL-34-M2 type group was significantly higher than that in the M2 type group(P＜0.05).Conclusion IL-34 can inhibit the polarization of macrophages derived from human monocytes cells towards M1 type,promote the polarization of macrophages towards M2 type,and enhance the migration ability of M2 type macrophages.
基金项目
江苏省"六大人才高峰"高层次人才资助项目(YY-177)
江苏省"青年医学人才"项目(QNRC2016400)
江苏省"333工程"项目(BRA2020196)
南通市科技局项目(MS22018007)
南通市重点医学人才项目(05)
江苏省研究生科研与实践创新计划项目(SJCX23_1807)
NETL
NSTL
万方数据