BACKGROUND Decreased circulating miR-197-3p was found in patients with recurrent deep vein thrombosis (DVT), but the specific role of miR-197-3p needs further exploration. MATERIALS AND METHODS Venous blood samples were collected from DVT patients and healthy controls, and peripheral blood mononuclear cells (PBMCs) were isolated to examine the expression patterns of miR-197-3p, CXCR2 and COX2 by qRT-PCR. Human umbilical vein endothelial cells (HUVECs) were further used as a cellular model to investigate the role of the miR-197-3p/CXCR2/COX2 axis in regulating cell viability, angiogenesis, and inflammation, which were determined by MTT assay, Matrigel-based tube formation assay, and enzyme-linked immunosorbent assay, respectively. Dual-luciferase reporter assay was used to examine the interactions between miR-198-3p and CXCR2. Expression of NF-κB p65 was examined by western blot to investigate whether the NF-κB pathway was involved in the regulatory effect of miR-197-3p on DVT. RESULTS miR-197-3p was decreased in PBMCs of patients with DVT, while CXCR2 and COX2 were increased compared to the healthy controls. In HUVECs, overexpression of miR-197-3p reduced CXCR2 levels and inhibited cell viability, angiogenesis, and release of inflammatory cytokines including TNF-α, IL-1β, and IL-6, which were reversed by miR-197-3p inhibition. Dual-luciferase reporter assay indicated miR-197-3p directly bound to CXCR2. CXCR2 further upregulated the expression of COX2 and activated the NF-κB pathway, promoting cell viability, angiogenesis and release of inflammatory cytokines in HUVECs. The effect of miR-197-3p inhibition on cell viability, angiogenesis and inflammation of HUVECs could be reversed by CXCR2 silencing. CONCLUSION MiR-197-3p affected viability, angiogenesis and inflammation of endothelial cells by targeting CXCR2/COX2 axis in vitro. Our findings provided a novel theoretical basis to investigate more effective therapies for DVT.

中文翻译：

---

MiR-197-3p 通过靶向 CXCR2/COX2 轴影响内皮细胞的血管生成和炎症。

背景在复发性深静脉血栓形成（DVT）患者中发现循环miR-​​197-3p降低，但miR-197-3p的具体作用需要进一步探索。材料与方法 采集 DVT 患者和健康对照者的静脉血样，分离外周血单个核细胞（PBMCs），通过 qRT-PCR 检测 miR-197-3p、CXCR2 和 COX2 的表达模式。人脐静脉内皮细胞 (HUVEC) 被进一步用作细胞模型，以研究 miR-197-3p/CXCR2/COX2 轴在调节细胞活力、血管生成和炎症中的作用，这些作用通过 MTT 测定、Matrigel-分别基于管形成测定和酶联免疫吸附测定。双荧光素酶报告基因检测用于检查 miR-198-3p 和 CXCR2 之间的相互作用。通过蛋白质印迹检查NF-κB p65的表达以研究NF-κB途径是否参与miR-197-3p对DVT的调节作用。结果 与健康对照组相比，DVT 患者 PBMC 中 miR-197-3p 降低，而 CXCR2 和 COX2 升高。在 HUVECs 中，miR-197-3p 的过表达降低了 CXCR2 水平并抑制了细胞活力、血管生成和包括 TNF-α、IL-1β 和 IL-6 在内的炎性细胞因子的释放，这些被 miR-197-3p 抑制所逆转。双荧光素酶报告基因检测表明 miR-197-3p 直接与 CXCR2 结合。CXCR2 进一步上调 COX2 的表达并激活 NF-κB 通路，促进 HUVECs 中的细胞活力、血管生成和炎性细胞因子的释放。miR-197-3p抑制对细胞活力的影响，HUVEC 的血管生成和炎症可以通过 CXCR2 沉默来逆转。结论 MiR-197-3p在体外通过靶向CXCR2/COX2轴影响内皮细胞的活力、血管生成和炎症。我们的研究结果为研究更有效的 DVT 疗法提供了新的理论基础。