巨噬细胞向肌成纤维细胞转变对矽肺的影响。

Effect of macrophage-to-myofibroblast transition on silicosis.

作者信息

Geng Fei, Xu Jingrou, Ren Xichen, Zhao Ying, Cai Yuhao, Li Yaqian, Jin Fuyu, Li Tian, Gao Xuemin, Cai Wenchen, Xu Hong, Wei Zhongqiu, Mao Na, Sun Ying, Yang Fang

机构信息

Hebei Key Laboratory for Organ Fibrosis Research, School of Public Health, North China University of Science and Technology, Tangshan, China.

Department of Pathology, Hebei Key Laboratory for Chronic Diseases, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan, China.

出版信息

Animal Model Exp Med. 2025 Feb;8(2):363-371. doi: 10.1002/ame2.12470. Epub 2024 Jul 9.

DOI:10.1002/ame2.12470

PMID:38979656

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11871087/

Abstract

BACKGROUND

The aim was to explore the effect of macrophage polarization and macrophage-to-myofibroblast transition (MMT) in silicosis.

METHODS

Male Wistar rats were divided into a control group and a silicosis group developed using a HOPE MED 8050 dynamic automatic dusting system. Murine macrophage MH-S cells were randomly divided into a control group and an SiO group. The pathological changes in lung tissue were observed using hematoxylin and eosin (HE) and Van Gieson (VG) staining. The distribution and location of macrophage marker (F4/80), M1 macrophage marker (iNOS), M2 macrophage marker (CD206), and myofibroblast marker (α-smooth muscle actin [α-SMA]) were detected using immunohistochemical and immunofluorescent staining. The expression changes in iNOS, Arg, α-SMA, vimentin, and type I collagen (Col I) were measured using Western blot.

RESULTS

The results of HE and VG staining showed obvious silicon nodule formation and the distribution of thick collagen fibers in the lung tissue of the silicosis group. Macrophage marker F4/80 increased gradually from 8 to 32 weeks after exposure to silica. Immunohistochemical and immunofluorescent staining results revealed that there were more iNOS-positive cells and some CD206-positive cells in the lung tissue of the silicosis group at 8 weeks. More CD206-positive cells were found in the silicon nodules of the lung tissues in the silicosis group at 32 weeks. Western blot analysis showed that the expressions of Inducible nitric oxide synthase and Arg protein in the lung tissues of the silicosis group were upregulated compared with those of the control group. The results of immunofluorescence staining showed the co-expression of F4/80, α-SMA, and Col I, and CD206 and α-SMA were co-expressed in the lung tissue of the silicosis group. The extracted rat alveolar lavage fluid revealed F4/80α-SMA, CD206α-SMA, and F4/80α-SMACol I cells using immunofluorescence staining. Similar results were also found in MH-S cells induced by SiO.

CONCLUSIONS

The development of silicosis is accompanied by macrophage polarization and MMT.

摘要

背景

旨在探讨巨噬细胞极化和巨噬细胞向肌成纤维细胞转变（MMT）在矽肺中的作用。

方法

将雄性Wistar大鼠分为对照组和使用HOPE MED 8050动态自动撒粉系统建立的矽肺组。将小鼠巨噬细胞MH-S细胞随机分为对照组和SiO组。采用苏木精-伊红（HE）和Van Gieson（VG）染色观察肺组织的病理变化。采用免疫组织化学和免疫荧光染色检测巨噬细胞标志物（F4/80）、M1巨噬细胞标志物（iNOS）、M2巨噬细胞标志物（CD206）和肌成纤维细胞标志物（α-平滑肌肌动蛋白[α-SMA]）的分布和定位。采用蛋白质免疫印迹法检测iNOS、精氨酸酶（Arg）、α-SMA、波形蛋白和I型胶原（Col I）的表达变化。

结果

HE和VG染色结果显示，矽肺组肺组织有明显的硅结节形成和粗大胶原纤维分布。暴露于二氧化硅后8至32周，巨噬细胞标志物F4/80逐渐增加。免疫组织化学和免疫荧光染色结果显示，矽肺组8周时肺组织中有较多的iNOS阳性细胞和一些CD206阳性细胞。矽肺组32周时肺组织硅结节中有更多的CD206阳性细胞。蛋白质免疫印迹分析显示，矽肺组肺组织中诱导型一氧化氮合酶和Arg蛋白的表达较对照组上调。免疫荧光染色结果显示，矽肺组肺组织中F4/80、α-SMA和Col I共表达，CD206和α-SMA共表达。采用免疫荧光染色从提取的大鼠肺泡灌洗液中检测到F4/80α-SMA、CD206α-SMA和F4/80α-SMACol I细胞。在SiO诱导的MH-S细胞中也发现了类似结果。