中华皮肤科杂志 ›› 2021, Vol. 54 ›› Issue (8): 688-695.doi: 10.35541/cjd.20210175

• 论著 • 上一篇    下一篇

梅毒螺旋体对巨噬细胞极化的诱导作用

张海妮1    寇彩霞1    刘进权1    张瑞华1    马印尼2    张瑞丽3    王千秋1   

  1. 1中国医学科学院、北京协和医学院皮肤病研究所临床防治室，南京  210042；2南京医科大学附属无锡第二医院皮肤科，无锡  214002；3南京医科大学第二附属医院皮肤科  210003 
  • 收稿日期:2021-03-03 修回日期:2021-04-30 发布日期:2021-08-02
  • 通讯作者: 王千秋；张瑞丽 E-mail:wangqianqiunj@126.com; reallyvictor@126.com
  • 基金资助:
    国家自然科学基金（81772209、81601804）；中国医学科学院医学与健康科技创新工程项目（2016-I2M-3021）；南京市国家级临床医学中心培育计划项目（2019060001）；无锡市科技发展资金项目（N20192021）

Inductive effect of Treponema pallidum on macrophage polarization

Zhang Haini1, Kou Caixia1, Liu Jinquan1, Zhang Ruihua1, Ma Yinni2, Zhang Ruili3, Wang Qianqiu1   

  1. 1Department of Sexually Transmitted Disease Clinical Management, Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042, China; 2Departmenet of Dermatology, Wuxi No.2 People′s Hospital Affiliated to Nanjing Medical University, Wuxi 214002, Jiangsu, China；3Department of Dermatology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing 210003, China
  • Received:2021-03-03 Revised:2021-04-30 Published:2021-08-02
  • Contact: Wang Qianqiu; Zhang Ruili E-mail:wangqianqiunj@126.com; reallyvictor@126.com
  • Supported by:
    National Natural Science Foundation of China （81772209, 81601804）; CAMS Innovation Fund for Medical Sciences （2016-I2M-3021）; Nanjing Incubation Program for National Clinical Research Center （2019060001）; Wuxi Science and Technology Development Fund Project （N20192021）

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摘要： 【摘要】 目的 探究巨噬细胞对梅毒螺旋体（Tp）的吞噬作用及Tp刺激后巨噬细胞的极化方向。方法 用Tp Nichols株作用人单核细胞THP-1来源的M0型巨噬细胞后，通过透射电镜和免疫荧光染色观察巨噬细胞对Tp的吞噬作用及巨噬细胞内结构的变化。Tp作用M0型巨噬细胞12 h后，继续培养24 h、48 h、72 h和6 d，分别通过Western印迹、免疫荧光染色观察M1型巨噬细胞表面标记CD86、M2型巨噬细胞表面标记CD163的表达，酶联免疫吸附试验检测巨噬细胞培养上清液中 M1型细胞因子白细胞介素（IL）-12 p70、干扰素γ、趋化因子配体10、IL-6、肿瘤坏死因子α和IL-1β水平，以及M2型细胞因子转化生长因子β1（TGF-β1）水平。采用Dunnett-t检验进行组间数据比较。结果 透射电镜观察发现，Tp刺激巨噬细胞后，巨噬细胞伸出伪足将Tp吞噬进细胞内，引起内质网肿胀、明显不规则增生和线粒体体积增大。而且，Tp刺激巨噬细胞后，继续培养24 h、48 h、72 h及6 d后巨噬细胞均高表达CD86，但低表达CD163，且24 h时上清液中IL-12 p70、干扰素γ、趋化因子配体10、IL-6、肿瘤坏死因子α和IL-1β均高于对照组（均P＜0.001），但TGF-β1与对照组差异无统计学意义（P＞0.05）。结论 THP-1来源的巨噬细胞吞噬Tp后内质网、线粒体结构发生变化；Tp可诱导M0型巨噬细胞极化为M1型巨噬细胞，且在6 d内不发生M1/M2型巨噬细胞再转化。

关键词: 苍白密螺旋体, 巨噬细胞活化, 细胞分化, 细胞因子类, 炎症

Abstract: 【Abstract】 Objective To investigate the phagocytosis of Treponema pallidum （Tp） by macrophages and the polarization direction of macrophages after Tp stimulation. Methods Human THP-1 monocyte-derived M0 macrophages were stimulated with the Tp Nichols strain, and the phagocytosis of Tp by macrophages and changes in the intracellular structure of macrophages were observed by transmission electron microscopy and immunofluorescence staining. After 12-hour stimulation by Tp, Tp was removed, the M0 macrophages continued to be cultured for 24, 48, 72 hours and 6 days. Western blot analysis and immunofluorescence staining were performed to determine the expression of the M1 macrophage marker CD86 and M2 macrophage marker CD163, and enzyme-linked immunosorbent assay was conducted to detect levels of M1-type cytokines interleukin （IL）-12 p70, interferon （IFN）- γ, chemokine ligand 10 （CXCL10）, IL-6, tumor necrosis factor （TNF）-α and IL-1β, as well as the M2-type cytokine transforming growth factor （TGF）-β1 in the culture supernatant of macrophages. Dunnett-t test was used for multiple comparisons. Results As transmission electron microscopy showed, after the stimulation by Tp, the macrophages extended pseudopodia and engulfed Tp, leading to swelling and obviously irregular hyperplasia of endoplasmic reticulum as well as enlargement of mitochondria. Moreover, after additional culture for 24, 48, 72 hours and 6 days, CD86 was highly expressed, but CD163 was lowly expressed in the Tp-treated macrophages; at 24 hours, the supernatant levels of IL-12 p70, IFN-γ, CXCL10, IL-6, TNF-α and IL-1β were significantly higher in the Tp-treated group than in the control group （all P＜0.001）, but there was no significant difference in the TGF-β1 supernatant level between the 2 groups （P＞0.05）. Conclusions After engulfment of Tp, the structures of endoplasmic reticulum and mitochondria in THP-1-derived macrophages markedly changed. Tp could induce the polarization of M0 macrophages into M1 macrophages, and phenotypic switch from M1 to M2 macrophage polarization was not observed within 6 days after Tp stimulation.

Key words: Treponema pallidum, Macrophage activation, Cell differentiation, Cytokines, Inflammation