Department of Infectious Disease, Yantai Mountain Hospital of Yantai, Yantai, Shandong, P. R. China.
Department of Respiratory and Critical Care Medicine, Zibo First Hospital, Zibo, Shandong, P. R. China.
Immunol Invest. 2022 May;51(4):787-801. doi: 10.1080/08820139.2021.1874977. Epub 2021 Jan 17.
Chronic obstructive pulmonary disease (COPD) is a chronic smoking-related lung disease associated with higher mortality and morbidity. Herein, we attempted to investigate the function of miR-558/TNF Receptor Superfamily Member 1A (TNFRSF1A) in the progression of COPD.
GEO database was applied to filtrate the differentially expressed mRNAs and miRNAs. KEGG enrichment was used to select the meaningful pathway related to the differentially expressed genes. TargetScan was used to predict the upstream regulator of TNFRSF1A, which was further affirmed by dual luciferase assay. HBE cells were stimulated by 20 μg/mL cigarette smoke extract (CSE) to mimic the COPD . The activity, apoptosis and inflammatory factors of HBE cells were evaluated by biological experiments. The levels of proteins related to TAK1/MAPK/NF-κB pathway were measured by Western blot.
TNFRSF1A is found to be highly expressed in COPD samples and enriched in TNF signaling pathway through bioinformatics analysis. miR-558 was verified as an upstream regulator of TNFRSF1A and negatively regulated TNFRSF1A expression. Up-regulation of miR-558 alleviated CSE-induced damage on HBE cells. The alleviative effect of miR-558 mimic on CSE-induced damage was suppressed by TNFRSF1A overexpression. The elevated expression of -TAK1p-p38 MAPK/p-NF-κB P65 in CSE condition was suppressed by miR-558 up-regulation. However, the results were reversed by TNFRSF1A overexpression. TAK1 inhibitor blocked the activation of TAK1/MAPK/NF-κB pathway, which was consistent with the results from miR-558 up-regulation.
Up-regulation of miR-558 relieved the damage of HBE cells-triggered by CSE via reducing TNFRSF1A and inactivating TAK1/MAPK/NF-κB pathway, affording novel molecules for COPD treatment.
慢性阻塞性肺疾病（COPD）是一种与较高死亡率和发病率相关的慢性吸烟相关肺部疾病。在此，我们试图研究 miR-558/肿瘤坏死因子受体超家族成员 1A（TNFRSF1A）在 COPD 进展中的作用。
应用 GEO 数据库筛选差异表达的 mRNAs 和 miRNAs。KEGG 富集分析用于选择与差异表达基因相关的有意义通路。TargetScan 用于预测 TNFRSF1A 的上游调控因子，并用双荧光素酶报告基因实验进一步验证。用 20μg/ml 香烟烟雾提取物（CSE）刺激 HBE 细胞模拟 COPD。通过生物学实验评估 HBE 细胞的活性、凋亡和炎症因子。用 Western blot 检测与 TAK1/MAPK/NF-κB 通路相关的蛋白水平。
通过生物信息学分析发现，TNFRSF1A 在 COPD 样本中高表达，并且在 TNF 信号通路中富集。miR-558 被验证为 TNFRSF1A 的上游调控因子，负调控 TNFRSF1A 的表达。上调 miR-558 可减轻 CSE 对 HBE 细胞的损伤。TNFRSF1A 过表达抑制了 miR-558 模拟物对 CSE 诱导损伤的缓解作用。上调 miR-558 可抑制 CSE 诱导的 TAK1p-p38 MAPK/p-NF-κB P65 表达升高。然而，当 TNFRSF1A 过表达时，结果相反。TAK1 抑制剂阻断了 TAK1/MAPK/NF-κB 通路的激活，这与 miR-558 上调的结果一致。
上调 miR-558 通过降低 TNFRSF1A 并使 TAK1/MAPK/NF-κB 通路失活，减轻 CSE 诱导的 HBE 细胞损伤，为 COPD 的治疗提供了新的分子靶点。
