Tian Xuan, Liu Jianlong, Jia Wei, Jiang Peng, Cheng Zhiyuan, Zhang Yunxin, Li Jinyong, Liu Xiao, Tian Chenyang
Department of Vascular Surgery, Beijing Jishuitan Hospital Beijing 100035, China.
Am J Transl Res. 2022 Jul 15;14(7):4666-4677. eCollection 2022.
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.
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.
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.
MiR-197-3p affected viability, angiogenesis and inflammation of endothelial cells by targeting CXCR2/COX2 axis . Our findings provided a novel theoretical basis to investigate more effective therapies for DVT.
在复发性深静脉血栓形成（DVT）患者中发现循环miR-197-3p水平降低，但miR-197-3p的具体作用尚需进一步探索。
采集DVT患者和健康对照者的静脉血样本，分离外周血单个核细胞（PBMC），采用qRT-PCR检测miR-197-3p、CXCR2和COX2的表达模式。进一步以人脐静脉内皮细胞（HUVEC）作为细胞模型，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，分别通过MTT法、基质胶基质管形成实验和酶联免疫吸附实验进行测定。采用双荧光素酶报告基因实验检测miR-198-3p与CXCR2之间的相互作用。通过蛋白质印迹法检测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。
与健康对照相比，DVT患者PBMC中miR-197-3p水平降低，而CXCR2和COX2水平升高。在HUVEC中，miR-197-3p过表达降低了CXCR2水平，并抑制了细胞活力、血管生成以及包括TNF-α、IL-1β和IL-6在内的炎性细胞因子的释放，miR-197-3p抑制可逆转这些作用。双荧光素酶报告基因实验表明miR-197-3p直接与CXCR2结合。CXCR2进一步上调COX2的表达并激活NF-κB途径，促进HUVEC的细胞活力、血管生成和炎性细胞因子的释放。CXCR2沉默可逆转miR-197-3p抑制对HUVEC细胞活力、血管生成和炎症的影响。
MiR-197-3p通过靶向CXCR2/COX2轴影响内皮细胞的活力、血管生成和炎症。我们的研究结果为探索更有效的DVT治疗方法提供了新的理论依据。
