糖皮质激素受体和 HDACs 在气道平滑肌细胞中的表达与 COPD 对类固醇的反应相关。

Expression of glucocorticoid receptor and HDACs in airway smooth muscle cells is associated with response to steroids in COPD.

机构信息

Department of Clinical Research, University Hospital Basel and University of Basel, Basel, Switzerland.

Clinic of Respiratory Medicine and Pulmonary Cell Research, University Hospital Basel, Basel, Switzerland.

出版信息

Respir Res. 2024 May 29;25(1):227. doi: 10.1186/s12931-024-02769-3.

DOI:10.1186/s12931-024-02769-3

PMID:38812021

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11137987/

Abstract

BACKGROUND

Steroid insensitivity in Chronic Obstructive Pulmonary Disease (COPD) presents a problem for controlling the chronic inflammation of the airways. The glucocorticoid receptor (GR) mediates the intracellular signaling of inhaled corticosteroids (ICS) by interacting with transcription factors and histone deacetylases (HDACs). The aim of this study was to assess if COPD patients' response to ICS in vivo, may be associated with the expression of GR, the complex of GR with transcription factors, and the expression of various HDACs in vitro.

METHODS

Primary airway smooth muscle cells (ASMC) were established from endobronchial biopsies obtained from patients with asthma (n = 10), patients with COPD (n = 10) and subjects that underwent diagnostic bronchoscopy without pathological findings and served as controls (n = 6). ASMC were also established from 18 COPD patients, 10 responders and 8 non-responders to ICS, who participated in the HISTORIC study, an investigator-initiated and driven clinical trial that proved the hypothesis that COPD patients with high ASMC in their endobronchial biopsies respond better to ICS than patients with low ASMC. Expression of GR and its isoforms GRα and GRβ and HDACs was investigated in primary ASMC in the absence or in the presence of dexamethasone (10M) by western blotting. The complex formation of GR with transcription factors was assessed by co-immunoprecipitation.

RESULTS

Expression of GR and its isoform GRα but not GRβ was significantly reduced in ASMC from COPD patients as compared to controls. There were no significant differences in the expression of GR, GRα and GRβ between responders and non-responders to ICS. However, treatment with dexamethasone upregulated the expression of total GR (p = 0.004) and GRα (p = 0.005) after 30 min in responders but not in non-responders. Τhe formation of the complex GR-c-Jun was increased 60 min after treatment with dexamethasone only in responders who exhibited significantly lower expression of HDAC3 (p = 0.005) and HDAC5 (p < 0.0001) as compared to non-responders.

CONCLUSIONS

These data suggest that ASMC from COPD patients who do not respond to treatment with ICS, are characterized by reduced GR-c-Jun complex formation and increased expression of HDAC3 and HDAC5.

TRIAL REGISTRATION

ISRCTN11017699 (Registration date: 15/11/2016).

摘要

背景

慢性阻塞性肺疾病（COPD）中的类固醇不敏感会导致气道慢性炎症的控制出现问题。糖皮质激素受体（GR）通过与转录因子和组蛋白去乙酰化酶（HDACs）相互作用，介导吸入性皮质类固醇（ICS）的细胞内信号转导。本研究旨在评估 COPD 患者体内对 ICS 的反应是否与 GR 的表达、GR 与转录因子的复合物以及各种 HDACs 的表达有关。

方法

从支气管内膜活检中建立原发性气道平滑肌细胞（ASMC），这些活检来自哮喘患者（n=10）、COPD 患者（n=10）和接受诊断性支气管镜检查但无病理发现的受试者（n=6）。ASMC 也来自 18 名 COPD 患者，其中 10 名对 ICS 有反应，8 名没有反应，他们参加了 HISTORIC 研究，这是一项由研究者发起和驱动的临床试验，证明了 COPD 患者支气管内膜活检中 ASMC 较高的患者对 ICS 的反应优于 ASMC 较低的患者。通过 Western blot 法在不存在或存在地塞米松（10M）的情况下，研究了原发性 ASMC 中 GR 及其同工型 GRα和 GRβ和 HDACs 的表达。通过共免疫沉淀评估 GR 与转录因子的复合物形成。

结果

与对照组相比，COPD 患者的 ASMC 中 GR 及其同工型 GRα的表达显著降低，但 GRβ的表达没有差异。对 ICS 有反应和无反应的患者之间 GR、GRα 和 GRβ 的表达没有差异。然而，地塞米松治疗 30 分钟后，仅在有反应的患者中，总 GR（p=0.004）和 GRα（p=0.005）的表达上调，但在无反应的患者中没有上调。地塞米松治疗 60 分钟后，GR-c-Jun 复合物的形成增加，仅在有反应的患者中，HDAC3（p=0.005）和 HDAC5（p<0.0001）的表达明显低于无反应的患者。