Department of Pharmacy, Jiangsu Health vocational college, Nanjing 211800, Jiangsu, China.
Department of Outpatient, Jurong People's Hospital, Zhenjiang 212400, Jiangsu, China.
Environ Toxicol. 2021 Sep;36(9):1775-1784. doi: 10.1002/tox.23298. Epub 2021 Jun 5.
Pneumonia is an inflammatory disease induced by infection with different pathogens. Currently, multiple preclinical studies have revealed that shikonin, a natural naphthoquinone, can mitigate lipopolysaccharide (LPS)-induced inflammation, but its underlying mechanism in pneumonia remains unknown. This research was designed to explore the function and regulatory mechanism of shikonin in LPS-induced cell injury and inflammation in WI-38 cells. In-vitro model of pneumonia was constructed by treating WI-38 cells with LPS. Expression of miR-489-3p and MAP2K1 was tested by RT-qPCR and (or) Western blot analysis. Cell viability was examined by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide assay. The productions of pro-inflammatory cytokines were determined by enzyme-linked immunosorbent assays. Cell apoptosis was detected by Western blot and flow cytometry analysis. In the current study, LPS induced WI-38 cell damage by inhibiting cell viability and promoting cell apoptosis and inflammation. Shikonin ameliorated LPS-induced cell injury and elevated miR-489-3p expression. LPS-induced inflammatory injury was further mitigated by upregulation of miR-489-3p. In addition, MAP2K1, the target of miR-489-3p, was upregulated by LPS. Furthermore, upregulation of MAP2K1 reversed the influence of shikonin and miR-489-3p mimics on LPS-induced cell injury and inflammation. This study revealed that shikonin protected WI-38 cells against LPS-induced cell injury and inflammatory response by regulating the miR-489-3p/MAP2K1 axis, thus affecting the progression of pneumonia.
肺炎是一种由不同病原体感染引起的炎症性疾病。目前，多项临床前研究表明，天然萘醌类化合物紫草素可以减轻脂多糖（LPS）诱导的炎症，但它在肺炎中的作用机制尚不清楚。本研究旨在探讨紫草素在 LPS 诱导的 WI-38 细胞损伤和炎症中的作用及其调控机制。通过用 LPS 处理 WI-38 细胞构建肺炎体外模型。采用 RT-qPCR 和（或）Western blot 分析检测 miR-489-3p 和 MAP2K1 的表达。通过 3-(4,5)-二甲基噻唑 (-z-y1)-3,5-二苯基四氮唑溴盐比色法检测细胞活力。通过酶联免疫吸附试验测定促炎细胞因子的产生。通过 Western blot 和流式细胞术分析检测细胞凋亡。在本研究中，LPS 通过抑制细胞活力、促进细胞凋亡和炎症来诱导 WI-38 细胞损伤。紫草素改善 LPS 诱导的细胞损伤，上调 miR-489-3p 的表达。上调 miR-489-3p 进一步减轻 LPS 诱导的炎症损伤。此外，LPS 上调了 miR-489-3p 的靶基因 MAP2K1。此外，MAP2K1 的上调逆转了紫草素和 miR-489-3p 模拟物对 LPS 诱导的细胞损伤和炎症的影响。本研究表明，紫草素通过调节 miR-489-3p/MAP2K1 轴来保护 WI-38 细胞免受 LPS 诱导的细胞损伤和炎症反应，从而影响肺炎的进展。
