ABSTRACT

Background

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.

Methods

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 in vitro. 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.

Results

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 p-TAK1/p-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.

Conclusions

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.

中文翻译：

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miR-558 通过靶向 TNFRSF1A 和灭活 TAK1/MAPK/NF-κB 通路减少暴露于香烟烟雾提取物的 HBE 细胞的损伤

摘要

背景

慢性阻塞性肺病 (COPD) 是一种与吸烟相关的慢性肺部疾病，与较高的死亡率和发病率有关。在此，我们试图研究 miR-558/TNF 受体超家族成员 1A (TNFRSF1A) 在 COPD 进展中的功能。

方法

GEO数据库用于过滤差异表达的mRNA和miRNA。KEGG富集用于选择与差异表达基因相关的有意义的途径。TargetScan用于预测TNFRSF1A的上游调节因子，双荧光素酶检测进一步证实了这一点。用 20 μg/mL 香烟烟雾提取物 (CSE) 刺激 HBE 细胞以在体外模拟 COPD 。通过生物学实验评价HBE细胞的活性、凋亡和炎症因子。Western blot检测TAK1/MAPK/NF-κB通路相关蛋白水平。

结果

通过生物信息学分析发现 TNFRSF1A 在 COPD 样本中高表达，并在 TNF 信号通路中富集。miR-558 被证实为 TNFRSF1A 的上游调节因子，并负调控 TNFRSF1A 的表达。miR-558的上调减轻了CSE诱导的HBE细胞损伤。TNFRSF1A 过表达抑制了 miR-558 模拟物对 CSE 诱导的损伤的缓解作用。在 CSE 条件下p - TAK1 / p-p38 MAPK/p-NF-κB P65的升高表达被 miR-558 上调抑制。然而，结果被 TNFRSF1A 过表达逆转。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。

结论

miR-558的上调通过减少TNFRSF1A和灭活TAK1/MAPK/NF-κB通路减轻了由CSE触发的HBE细胞损伤，为COPD治疗提供了新的分子。