Regulatory mechanism of DDX5 in ox-LDL-induced endothelial cell injury through the miR-640/SOX6 axis.

Clinical hemorheology and microcirculation Pub Date : 2024-01-01 DOI:10.3233/CH-242254

Shuo Li, Yu Wang

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Abstract

Background: Endothelial dysfunction is an early and pre-clinical manifestation of coronary heart disease (CHD).

Objective: This study investigates the role of DDX5 in oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell injury to confer novel targets for the treatment of CHD.

Methods: Endothelial cells were induced by ox-LDL. DDX5, pri-miR-640, pre-miR-640, miR-640, and SOX6 expressions were analyzed by RT-qPCR and Western blot. DDX5 expression was intervened by shRNA, followed by CCK-8 analysis of proliferation, flow cytometry detection of apoptosis, and tube formation assay analysis of angiogenic potential of cells. The binding between DDX5 and pri-miR-640 was determined by RIP, and the pri-miR-640 RNA stability was measured after actinomycin D treatment. Dual-luciferase assay verified the targeting relationship between miR-640 and SOX6.

Results: DDX5 and miR-640 were highly expressed while SOX6 was poorly expressed in ox-LDL-induced endothelial cells. Silence of DDX5 augmented cell proliferation, abated apoptosis, and facilitated angiogenesis. Mechanistically, RNA binding protein DDX5 elevated miR-640 expression by weakening the degradation of pri-miR-640, thereby reducing SOX6 expression. Combined experimental results indicated that overexpression of miR-640 or low expression of SOX6 offset the protective effect of DDX5 silencing on cell injury.

Conclusion: DDX5 elevates miR-640 expression by repressing the degradation of pri-miR-640 and then reduces SOX6 expression, thus exacerbating ox-LDL-induced endothelial cell injury.

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DDX5 通过 miR-640/SOX6 轴在氧化-LDL 诱导的内皮细胞损伤中的调控机制

背景：内皮功能障碍是冠心病（CHD）的早期和临床前表现：内皮功能障碍是冠心病（CHD）的早期和临床前表现：本研究探讨了DDX5在氧化低密度脂蛋白（ox-LDL）诱导的内皮细胞损伤中的作用，从而为治疗冠心病提供新的靶点：方法：用氧化-LDL诱导内皮细胞。方法：用 ox-LDL 诱导内皮细胞，通过 RT-qPCR 和 Western 印迹分析 DDX5、pri-miR-640、pre-miR-640、miR-640 和 SOX6 的表达。用 shRNA 干预 DDX5 的表达，然后用 CCK-8 分析细胞的增殖情况，用流式细胞仪检测细胞的凋亡情况，用试管形成试验分析细胞的血管生成潜能。用 RIP 法测定了 DDX5 与 pri-miR-640 的结合情况，并测定了放线菌素 D 处理后 pri-miR-640 RNA 的稳定性。双荧光素酶检测验证了 miR-640 与 SOX6 之间的靶向关系：结果：在氧化-LDL 诱导的内皮细胞中，DDX5 和 miR-640 高表达，而 SOX6 低表达。沉默 DDX5 可促进细胞增殖、减少细胞凋亡并促进血管生成。从机理上讲，RNA 结合蛋白 DDX5 通过削弱 pri-miR-640 的降解来提高 miR-640 的表达，从而降低 SOX6 的表达。综合实验结果表明，miR-640的过度表达或SOX6的低表达抵消了DDX5沉默对细胞损伤的保护作用：结论：DDX5通过抑制pri-miR-640的降解来提高miR-640的表达，进而降低SOX6的表达，从而加剧氧化-LDL诱导的内皮细胞损伤。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Clinical hemorheology and microcirculation

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