CH25H/25-HC promotes pulmonary fibrosis via AMPK/STAT6 pathway-dependent M2 macrophage polarization in COPD

IF 2.5 4区 医学 Q3 IMMUNOLOGY
Ying Li, Guangzhi Xiao, Xianghui Fu, Xing Luo, Fengfan Yang, Yadan Li, Zhaohui Zheng
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Abstract

Objective

Chronic obstructive pulmonary disease (COPD) is intricately linked to pulmonary fibrosis, yet the underlying mechanisms remain unclear. This study investigates whether CH25H/25-hydroxycholesterol (25-HC) promotes pulmonary fibrosis in COPD by modulating AMPK/STAT6-dependent M2 macrophage polarization.

Methods

Using GEO datasets and a cigarette smoke-induced COPD mouse model, we analyzed CH25H expression and fibrotic pathology. CH25H was silenced via adeno-associated virus (AAV)-delivered shRNA. Histopathology, flow cytometry, qPCR, and Western blotting assessed fibrosis, macrophage polarization (M1/M2), and AMPK/STAT6 pathway activity. Bone marrow-derived macrophages (BMDMs) were employed to validate polarization mechanisms. The role of the AMPK/STAT6 pathway was investigated using the specific activator.

Results

Analysis of the GEO database and experimental verification showed significantly increased CH25H expression in both lung tissues and macrophages from COPD mice. CH25H knockdown alleviated alveolar damage, airway remodeling, and pulmonary fibrosis in COPD mice, evidenced by reduced expression of fibrosis-related proteins, improved lung function, and attenuated inflammatory responses. Moreover, CH25H knockdown inhibited M2 macrophage polarization and decreased the proportion of M2-type macrophages. Importantly, decreased levels of 25-HC following CH25H knockdown were asso ciated with suppressed activation of the AMPK/STAT6 pathway. Rescue experiments demonstrated that the protective effects of CH25H knockdown could be reversed by adding the AMPKα activator GSK621.

Conclusion

Our findings demonstrate that CH25H/25-HC exacerbates COPD-associated pulmonary fibrosis by promoting AMPK/STAT6-dependent M2 macrophage polarization. Targeting CH25H may represent a novel therapeutic strategy for mitigating fibrosis in COPD.
CH25H/25-HC通过AMPK/STAT6通路依赖的M2巨噬细胞极化促进COPD肺纤维化
慢性阻塞性肺疾病(COPD)与肺纤维化有着复杂的联系,但其潜在机制尚不清楚。本研究探讨CH25H/25-羟基胆固醇(25-HC)是否通过调节AMPK/ stat6依赖性M2巨噬细胞极化促进COPD肺纤维化。方法使用GEO数据集和香烟烟雾诱导的COPD小鼠模型,分析CH25H的表达和纤维化病理。通过腺相关病毒(AAV)递送shRNA使CH25H沉默。组织病理学、流式细胞术、qPCR和Western blotting评估纤维化、巨噬细胞极化(M1/M2)和AMPK/STAT6通路活性。骨髓源性巨噬细胞(bmdm)被用来验证极化机制。使用特异性激活剂研究AMPK/STAT6通路的作用。结果GEO数据库分析和实验验证显示,慢性阻塞性肺病小鼠肺组织和巨噬细胞中CH25H表达均显著升高。CH25H敲低可减轻COPD小鼠的肺泡损伤、气道重塑和肺纤维化,这可以通过纤维化相关蛋白的表达降低、肺功能改善和炎症反应减弱来证明。CH25H敲低抑制M2型巨噬细胞极化,降低M2型巨噬细胞比例。重要的是,CH25H敲除后25-HC水平的降低与AMPK/STAT6通路的抑制激活有关。救援实验表明,添加AMPKα激活剂GSK621可以逆转CH25H敲低的保护作用。结论CH25H/25-HC通过促进AMPK/ stat6依赖性M2巨噬细胞极化,加重copd相关肺纤维化。靶向CH25H可能是缓解COPD纤维化的一种新的治疗策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Immunobiology
Immunobiology 医学-免疫学
CiteScore
5.00
自引率
3.60%
发文量
108
审稿时长
55 days
期刊介绍: Immunobiology is a peer-reviewed journal that publishes highly innovative research approaches for a wide range of immunological subjects, including • Innate Immunity, • Adaptive Immunity, • Complement Biology, • Macrophage and Dendritic Cell Biology, • Parasite Immunology, • Tumour Immunology, • Clinical Immunology, • Immunogenetics, • Immunotherapy and • Immunopathology of infectious, allergic and autoimmune disease.
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