Lu Wei, Wan Guoqing, Zhu He, Zhu Tao, Zhang Xinmei
Department of Cardiovascular Surgery, The Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People's Hospital, Quzhou, 324000, China.
School of Pharmacy, Shanghai University of Medicine and Health Sciences, Shanghai, China.
Heliyon. 2024 Mar 27;10(7):e28887. doi: 10.1016/j.heliyon.2024.e28887. eCollection 2024 Apr 15.
The impairment of endothelial cells triggered by oxidized low-density lipoprotein (ox-LDL) stands as a critical event in the advancement of atherosclerosis (AS). MiR-497-5p has been recognized as a potential predictor for AS, but its precise involvement in ox-LDL-induced endothelial cell dysfunction remains to be elucidated.
An AS cell model was established by exposing human umbilical vein endothelial cells (HUVECs) to 100 μg/mL ox-LDL for 24 h. The assessment of endothelial cell function included evaluating cell viability, caspase-3 activity, inflammatory factors, and oxidative markers. Molecular mechanisms were elucidated through quantitative real-time PCR, Western blot analysis, and luciferase reporter assays.
Our investigation revealed that exposure to ox-LDL led to an upregulation in miR-497-5p and p-p38 levels, while downregulating the expression of vascular endothelial growth factor A (VEGFA) and phosphorylated (p)-endothelial nitric oxide synthase (-eNOS) in HUVECs. Ox-LDL exposure resulted in decreased cell viability and angiogenic capacity, coupled with increased apoptosis, inflammation, and oxidative stress in HUVECs, partially mediated by the upregulation of miR-497-5p. We confirmed VEGFA as a direct target of miR-497-5p. Interfering with VEGFA expression significantly reversed the effects mediated by miR-497-5p silencing in HUVECs exposed to ox-LDL.
In summary, our findings demonstrate that miR-497-5p exacerbates ox-LDL-induced dysfunction in HUVECs through the activation of the p38/MAPK pathway, mediated by the targeting of VEGFA.
氧化型低密度脂蛋白（ox-LDL）引发的内皮细胞损伤是动脉粥样硬化（AS）进展中的关键事件。MiR-497-5p已被认为是AS的潜在预测指标，但其在ox-LDL诱导的内皮细胞功能障碍中的具体作用仍有待阐明。
通过将人脐静脉内皮细胞（HUVECs）暴露于100μg/mL ox-LDL中24小时建立AS细胞模型。内皮细胞功能评估包括评估细胞活力、半胱天冬酶-3活性、炎症因子和氧化标志物。通过定量实时PCR、蛋白质免疫印迹分析和荧光素酶报告基因检测阐明分子机制。
我们的研究表明，暴露于ox-LDL会导致HUVECs中miR-497-5p和p-p38水平上调，同时下调血管内皮生长因子A（VEGFA）和磷酸化（p）-内皮型一氧化氮合酶（-eNOS）的表达。ox-LDL暴露导致HUVECs细胞活力和血管生成能力下降，同时细胞凋亡、炎症和氧化应激增加，部分是由miR-497-5p上调介导的。我们证实VEGFA是miR-497-5p的直接靶点。干扰VEGFA表达可显著逆转miR-497-5p沉默在暴露于ox-LDL的HUVECs中所介导的作用。
总之，我们的研究结果表明，miR-497-5p通过靶向VEGFA激活p38/MAPK途径，加剧了ox-LDL诱导的HUVECs功能障碍。
