Department of Medicine, College of Medicine, Gachon University, Incheon, Republic of Korea.
Department of Allergy, Pulmonary and Critical Care Medicine, Gachon University, Gil Medical Center, Incheon, Republic of Korea.
Toxicol In Vitro. 2022 Oct;84:105428. doi: 10.1016/j.tiv.2022.105428. Epub 2022 Jul 4.
Particulate matter (PM) is associated with the incidence, exacerbation, and mortality of variable respiratory diseases. However, the molecular mechanisms of PM-mediated inflammation are unclear. We identified microRNAs (miRNAs) and messenger RNAs (mRNAs) related to the inflammatory response in PM-exposed bronchial epithelial cells using next-generation sequencing. Of the miRNAs, miR-6515-5p was significantly downregulated in PM-exposed human bronchial epithelial BEAS-2B cells. miR-6515-5p regulated the production of pro-inflammatory cytokines (IL-6 and IL-8) and the expression of inflammatory genes (IL-1β, IL-6, IL-8, TNF-α, CXCL-1, and MCP-1) via MAPK/ERK signaling; overexpression of miR-6515-5p using a mimic inhibited PM-induced inflammatory responses via inactivation of the ERK pathway, whereas downregulation of miR-6515-5p via an inhibitor significantly increased inflammation in PM-exposed cells via activation of ERK. Furthermore, we identified colony stimulating factor 3 (CSF3) as a target gene of miR-6515-5p using TargetScanHuman, and confirmed the association between miR-6515-5p and CSF3 using a luciferase reporter assay. Furthermore, we found that mRNA and protein levels of CSF3 were negatively regulated by miR-6515-5p. Inhibition of CSF3 by small interfering RNA significantly reduced the expression and production of inflammatory markers in PM-exposed cells by inactivating the MAPK/ERK signaling pathway. Therefore, we suggest that miR-6515-5p regulates PM-induced inflammatory responses by targeting CSF3 via MAPK/ERK signaling in bronchial epithelial cells.
颗粒物 (PM) 与各种呼吸道疾病的发病率、恶化和死亡率有关。然而，PM 介导的炎症的分子机制尚不清楚。我们使用下一代测序技术鉴定了暴露于 PM 的支气管上皮细胞中与炎症反应相关的 microRNAs (miRNAs) 和信使 RNAs (mRNAs)。在 miRNAs 中，miR-6515-5p 在暴露于 PM 的人支气管上皮 BEAS-2B 细胞中显著下调。miR-6515-5p 通过 MAPK/ERK 信号调节促炎细胞因子 (IL-6 和 IL-8) 的产生和炎症基因 (IL-1β、IL-6、IL-8、TNF-α、CXCL-1 和 MCP-1) 的表达；通过模拟物过表达 miR-6515-5p 通过失活 ERK 途径抑制 PM 诱导的炎症反应，而通过抑制剂下调 miR-6515-5p 通过激活 ERK 显着增加暴露于 PM 的细胞中的炎症。此外，我们使用 TargetScanHuman 鉴定了集落刺激因子 3 (CSF3) 作为 miR-6515-5p 的靶基因，并通过荧光素酶报告基因检测证实了 miR-6515-5p 与 CSF3 之间的关联。此外，我们发现 CSF3 的 mRNA 和蛋白水平受 miR-6515-5p 的负调控。通过小干扰 RNA 抑制 CSF3 通过失活 MAPK/ERK 信号通路显着降低暴露于 PM 的细胞中炎症标志物的表达和产生。因此，我们认为 miR-6515-5p 通过 MAPK/ERK 信号通路靶向 CSF3 调节支气管上皮细胞中 PM 诱导的炎症反应。
