Abstract

Background

Bronchopneumonia is a disease of the respiratory tract. It leads to other complications and endangers life and health. Long non-coding RNA (lncRNA) participates in the occurrence and development of bronchopneumonia. Nuclear paraspeckle assembly transcript 1 (NEAT1) plays a key role in inflammatory diseases, but the function of NEAT1 in bronchopneumonia remains unclear.

Methods

RT-qPCR and Western blotting were performed to determine genes and proteins expressions. MTT was applied to test cell viability. Cell apoptosis was detected by flow cytometry. RIP was used to investigate the correlation between NEAT1 and miR-155-5p. The interaction between miR-155-5p and NEAT1 or MyD88 was evaluated by the dual-luciferase reporter gene.

Results

NEAT1 and MyD88 were upregulated in BEAS-2B cells by LPS, while miR-155-5p was downregulated. Knockdown of NEAT1 inhibited LPS-induced BEAS-2B cells growth inhibition by inhibiting the apoptosis. In addition, NEAT1 silencing suppressed LPS-induced inflammatory responses in BEAS-2B cells via suppression of TNF-α, IL-1β, IL-6, and IL-18. Meanwhile, NEAT1 is directly bound to miR-155-5p to regulate MyD88/NF-κB axis, and overexpression of miR-155-5p increased cell proliferation and suppressed inflammatory factors expression levels and cell apoptosis. Furthermore, sh-NEAT1-induced inhibition of BEAS-2B cells injury was partially reversed by miR-155-5p inhibitor or MyD88 overexpression.

Conclusion

NEAT1 silencing suppressed LPS-induced BEAS-2B cells injury and inflammation by the mediation of miR-155-5p/MyD88/NF-κB axis. Thus, our study might shed new light on exploring the new strategies for the treatment of bronchopneumonia.

中文翻译：

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LncRNA NEAT1通过靶向miR-155-5p激活支气管肺炎中的MyD88/NF-κB通路

摘要

背景

支气管肺炎是一种呼吸道疾病。它会导致其他并发症并危及生命和健康。长链非编码RNA（lncRNA）参与支气管肺炎的发生发展。核旁斑点组装转录物 1 (NEAT1) 在炎症性疾病中起关键作用，但 NEAT1 在支气管肺炎中的功能仍不清楚。

方法

进行 RT-qPCR 和蛋白质印迹以确定基因和蛋白质的表达。MTT用于测试细胞活力。通过流式细胞术检测细胞凋亡。RIP 用于研究 NEAT1 和 miR-155-5p 之间的相关性。miR-155-5p 与 NEAT1 或 MyD88 之间的相互作用通过双荧光素酶报告基因进行评估。

结果

NEAT1 和 MyD88 在 BEAS-2B 细胞中被 LPS 上调，而 miR-155-5p 被下调。NEAT1 的敲低通过抑制细胞凋亡来抑制 LPS 诱导的 BEAS-2B 细胞生长抑制。此外，NEAT1 沉默通过抑制 TNF-α、IL-1β、IL-6 和 IL-18 来抑制 BEAS-2B 细胞中 LPS 诱导的炎症反应。同时，NEAT1直接与miR-155-5p结合以调节MyD88/NF-κB轴，miR-155-5p的过表达增加细胞增殖并抑制炎症因子表达水平和细胞凋亡。此外，sh-NEAT1 诱导的 BEAS-2B 细胞损伤抑制被 miR-155-5p 抑制剂或 MyD88 过表达部分逆转。

结论

NEAT1 沉默通过 miR-155-5p/MyD88/NF-κB 轴的介导抑制 LPS 诱导的 BEAS-2B 细胞损伤和炎症。因此，我们的研究可能会为探索治疗支气管肺炎的新策略提供新的思路。