Nursing School, Zheng Zhou Railway Vocational and Technical College, Zhengzhou, 451460, Henan, China.
Department of Medical Nursing, College of Nursing, Hebei University of Chinese Medicine, Shijiazhuang, 050200, Hebei, China.
Inflamm Res. 2022 Jun;71(5-6):565-576. doi: 10.1007/s00011-022-01576-0. Epub 2022 Apr 30.
Chronic obstructive pulmonary disease (COPD) is a chronic airway disease with airflow limitation and abnormal inflammatory response. It has been verified that SOX9 plays a key role in lung function of various lung diseases and SOX9 is closely associated with COPD. Additionally, literature has reported that STIM1 is involved in lung injury and is highly expressed in neutrophils from COPD patients. This study aimed to characterize the biological roles of SOX9 and STIM1 in the pathogenesis of COPD and to elucidate the regulatory mechanism.
Human bronchial epithelial cells (BEAS-2B) were treated with CSE to construct in vitro COPD model. The levels of SOX9 and STIM1 in CSE-treated BEAS-2B cells were detected by western blot and RT-qPCR assay. Then, JASPAR datasets were utilized to analyze SOX9 binding sites in the promoter region of STIM1. Besides, luciferase reporter assay and ChIP assay were employed to validate the binding sites in STIM1 promoter region to SOX9. In addition, viability and apoptosis of BEAS-2B cells were assessed by utilizing MTT assay and TUNEL staining. ELISA kits and corresponding commercial kits were applied to measure the levels of TNF-α, IL-6, IL-1β, SOD, GSH-Px and MDA.
CSE treatment dose- and time-dependently reduced SOX9 expression in BEAS-2B cells. SOX9 overexpression enhanced the viability and suppressed the apoptosis of CSE-treated BEAS-2B cells as well as attenuated CSE-induced inflammation and oxidative stress. Then, it was validated that SOX9 bound to the promoter region of STIM1. Moreover, SOX9 overexpression-mediated impacts on cell viability, cell apoptosis, inflammation and oxidative stress in CSE-treated BEAS-2B cells were partially abolished by upregulation of STIM1.
To sum up, results here suggested that overexpression of SOX9 could mitigate inflammatory injury in CSE-treated bronchial epithelial cells by suppressing STIM1.
慢性阻塞性肺疾病（COPD）是一种以气流受限和异常炎症反应为特征的慢性气道疾病。已有研究证实，SOX9 在各种肺部疾病的肺功能中发挥关键作用，并且与 COPD 密切相关。此外，文献报道 STIM1 参与肺损伤，并且在 COPD 患者的中性粒细胞中高度表达。本研究旨在阐明 SOX9 和 STIM1 在 COPD 发病机制中的生物学作用及其调控机制。
用 CSE 处理人支气管上皮细胞（BEAS-2B）构建体外 COPD 模型。通过 Western blot 和 RT-qPCR 检测 CSE 处理的 BEAS-2B 细胞中 SOX9 和 STIM1 的水平。然后，利用 JASPAR 数据集分析 SOX9 在 STIM1 启动子区域的结合位点。此外，还利用荧光素酶报告基因检测和 ChIP 检测来验证 STIM1 启动子区域与 SOX9 的结合位点。另外，通过 MTT 检测和 TUNEL 染色评估 BEAS-2B 细胞的活力和凋亡。采用 ELISA 试剂盒和相应的商业试剂盒测定 TNF-α、IL-6、IL-1β、SOD、GSH-Px 和 MDA 的水平。
CSE 处理呈剂量和时间依赖性地降低 BEAS-2B 细胞中 SOX9 的表达。SOX9 过表达增强了 CSE 处理的 BEAS-2B 细胞的活力，抑制了细胞凋亡，并减轻了 CSE 诱导的炎症和氧化应激。然后，验证了 SOX9 与 STIM1 的启动子区域结合。此外，SOX9 过表达介导的对 CSE 处理的 BEAS-2B 细胞中细胞活力、细胞凋亡、炎症和氧化应激的影响，部分被 STIM1 的上调所消除。
总之，结果表明，SOX9 的过表达通过抑制 STIM1 减轻 CSE 处理的支气管上皮细胞中的炎症损伤。
