Abstract Circulating miR-150-5p has been identified as a prognostic marker in patients with critical illness and sepsis. Herein, we aimed to further explore the role and underlying mechanism of miR-150-5p in sepsis. Quantitative real-time-PCR assay was performed to detect the expression of miR-150-5p upon stimulation with lipopolysaccharide (LPS) in RAW264.7 cells. The levels of tumor necrosis factor-α, interleukin (IL)-6 and IL-1β were measured by ELISA assay. Cell apoptosis was determined using flow cytometry. Western blot was used to assess notch receptor 1 (Notch1) expression in LPS-induced RAW264.7 cells. Dual-luciferase reporter assay was employed to validate the target of miR-150-5p. Our data showed that miR-150-5p was downregulated and Notch1 was upregulated in LPS-stimulated RAW264.7 cells. miR-150-5p overexpression or Notch1 silencing alleviated LPS-induced inflammatory response and apoptosis in RAW264.7 cells. Moreover, Notch1 was a direct target of miR-150-5p. Notch1 abated miR-150-5p-mediated anti-inflammation and anti-apoptosis in LPS-induced RAW264.7 cells. miR-150-5p alleviated LPS-induced inflammatory response and apoptosis at least partly by targeting Notch1 in RAW264.7 cells, highlighting miR-150-5p as a target in the development of anti-inflammation and anti-apoptosis drugs for sepsis treatment.

中文翻译：

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miR-150-5p的上调通过靶向Notch1减轻LPS诱导的RAW264.7巨噬细胞炎症反应和凋亡

摘要 循环 miR-150-5p 已被确定为危重病和败血症患者的预后标志物。在此，我们旨在进一步探索 miR-150-5p 在脓毒症中的作用和潜在机制。进行定量实时 PCR 测定以检测 RAW264.7 细胞中脂多糖 (LPS) 刺激后 miR-150-5p 的表达。ELISA法测定肿瘤坏死因子-α、白细胞介素(IL)-6和IL-1β水平。使用流式细胞术测定细胞凋亡。蛋白质印迹用于评估 LPS 诱导的 RAW264.7 细胞中的 Notch 受体 1 (Notch1) 表达。双荧光素酶报告基因检测用于验证 miR-150-5p 的靶标。我们的数据显示，在 LPS 刺激的 RAW264.7 细胞中，miR-150-5p 下调，Notch1 上调。miR-150-5p 过表达或 Notch1 沉默减轻了 LPS 诱导的 RAW264.7 细胞炎症反应和细胞凋亡。此外，Notch1 是 miR-150-5p 的直接靶标。Notch1 在 LPS 诱导的 RAW264.7 细胞中减弱了 miR-150-5p 介导的抗炎和抗细胞凋亡。miR-150-5p 至少部分通过靶向 RAW264.7 细胞中的 Notch1 减轻 LPS 诱导的炎症反应和细胞凋亡，突出 miR-150-5p 作为开发用于脓毒症治疗的抗炎和抗凋亡药物的靶点。