Department of Clinical Laboratory Sciences, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, P.R. China.
Translational Medical Center, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, P.R. China.
Sci Prog. 2021 Apr-Jun;104(2):368504211014361. doi: 10.1177/00368504211014361.
Acute lung injury (ALI) is one of the most prevalent respiratory syndromes of excessive inflammatory reaction during lung infection. () infection is among the leading causes of ALI. MicroRNAs (miRNAs) regulate the expression of target mRNAs, including those involved in inflammatory processes, by binding to the 3'UTR. To date, the roles of miRNAs in -induced ALI remain unclear. In this study, we investigated the role of miR-384-5p in -induced ALI and its underlying molecular mechanism. RT-PCR, Western blot, ELISA, Myeloperoxidase (MPO) assay, microRNA target analysis, transient transfection, and luciferase reporter assay were utilized. In vivo study was conducted on mouse model. The expression of miR-384-5p was upregulated and positively correlated with inflammatory cytokine production in lung tissues and RAW264.7 and J774A.1 macrophages infected with . The miR-384-5p inhibitor alleviated the inflammatory reaction induced by . Target prediction analysis revealed that PGC1β was a target of miR-384-5p, which was further validated by the PGC1β 3'-UTR luciferase assay and the inverse correlation between the expression of miR-384-5p and PGC1β in -infected ALI tissues and macrophages. Moreover, macrophages transfected with miR-384-5p mimic exhibited reduced levels of PGC1β. The suppression of the expression of PGC1β by infection in the macrophages was abrogated by miR-384-5p inhibitor. Then, we demonstrated that PGC1β played an inhibitory role in -induced production of inflammatory cytokines. Furthermore, suppression of miR-384-5p in macrophages inhibited the activation of the NF-κB, MAPK, and Akt signaling pathways triggered by , but not the STAT3 pathway. These results demonstrate that miR-384-5p contributes to -induced ALI at least in part by targeting PGC1β and enhancing the activation of the NF-κB, MAPK, and Akt inflammatory signaling pathways. Thus, targeting miR-384-5p might exert a protective effect on -induced ALI.
急性肺损伤（ALI）是肺部感染时过度炎症反应最常见的呼吸系统综合征之一。（）感染是 ALI 的主要原因之一。microRNAs（miRNAs）通过与 3'UTR 结合来调节靶 mRNA 的表达，包括参与炎症过程的靶 mRNA。迄今为止，miRNAs 在 - 诱导的 ALI 中的作用尚不清楚。在这项研究中，我们研究了 miR-384-5p 在 - 诱导的 ALI 中的作用及其潜在的分子机制。我们使用了 RT-PCR、Western blot、ELISA、髓过氧化物酶（MPO）测定、microRNA 靶分析、瞬时转染和荧光素酶报告基因测定。在小鼠模型中进行了体内研究。在感染的肺组织和 RAW264.7 和 J774A.1 巨噬细胞中，miR-384-5p 的表达上调，并与炎症细胞因子的产生呈正相关。miR-384-5p 抑制剂减轻了由诱导的炎症反应。靶预测分析表明 PGC1β 是 miR-384-5p 的靶标，通过 PGC1β 3'-UTR 荧光素酶测定和在感染的 ALI 组织和巨噬细胞中 miR-384-5p 和 PGC1β 表达之间的负相关得到进一步验证。此外，转染 miR-384-5p 模拟物的巨噬细胞中 PGC1β 的表达水平降低。PGC1β 表达的抑制在感染的巨噬细胞中被 miR-384-5p 抑制剂所阻断。然后，我们证明 PGC1β 在 - 诱导的炎症细胞因子产生中发挥抑制作用。此外，巨噬细胞中 miR-384-5p 的抑制抑制了，但不抑制 STAT3 途径，由诱导的 NF-κB、MAPK 和 Akt 炎症信号通路的激活。这些结果表明，miR-384-5p 通过靶向 PGC1β 并增强 NF-κB、MAPK 和 Akt 炎症信号通路的激活，至少部分导致 - 诱导的 ALI。因此，靶向 miR-384-5p 可能对 - 诱导的 ALI 发挥保护作用。
