Department of Cardiology, The Puren Hospital, No. 218, Changqing First Road, Jianghan District, Wuhan, 430081, Hubei, China.
BMC Cardiovasc Disord. 2021 Apr 23;21(1):207. doi: 10.1186/s12872-021-02012-7.
Circular RNAs have shown important regulatory roles in cardiovascular diseases, containing atherosclerosis (AS). We intended to explore the role of circ_0004104 in AS using oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cells and its associated mechanism.
Real-time quantitative polymerase chain reaction and Western blot assay were conducted to analyze RNA levels and protein levels, respectively. Cell viability, apoptosis, angiogenic ability and inflammatory response were assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, flow cytometry, capillary-like network formation assay and enzyme-linked immunosorbent assay, respectively. Cell oxidative stress was assessed using commercial kits. Dual-luciferase reporter assay, RNA immunoprecipitation assay and RNA-pull down assay were performed to verify the intermolecular interaction.
ox-LDL exposure up-regulated the level of circ_0004104 in HUVECs. ox-LDL exposure suppressed cell viability and angiogenic ability whereas promoted the apoptosis, inflammation and oxidative stress of HUVECs partly through up-regulating circ_0004104. MicroRNA-328-3p (miR-328-3p) was confirmed as a target of circ_0004104. MiR-328-3p interference largely reversed circ_0004104 silencing-mediated effects in HUVECs upon ox-LDL exposure. MiR-328-3p interacted with the 3' untranslated region of tripartite motif 14, and circ_0004104 positively regulated TRIM14 expression by sponging miR-328-3p. TRIM14 overexpression largely overturned miR-328-3p accumulation-induced influences in HUVECs upon ox-LDL exposure.
Circ_0004104 knockdown attenuated ox-LDL-induced dysfunction in HUVECs via miR-328-3p-mediated regulation of TRIM14.
环状 RNA 在心血管疾病中具有重要的调控作用，包含动脉粥样硬化（AS）。我们旨在使用氧化低密度脂蛋白（ox-LDL）诱导的血管内皮细胞来研究 circ_0004104 在 AS 中的作用及其相关机制。
实时定量聚合酶链反应和 Western blot 分析分别用于分析 RNA 水平和蛋白水平。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定、流式细胞术、毛细血管样网络形成测定和酶联免疫吸附测定分别评估细胞活力、凋亡、血管生成能力和炎症反应。使用商业试剂盒评估细胞氧化应激。双荧光素酶报告基因测定、RNA 免疫沉淀测定和 RNA 下拉测定用于验证分子间相互作用。
ox-LDL 暴露上调了 HUVECs 中 circ_0004104 的水平。ox-LDL 暴露抑制了细胞活力和血管生成能力，而促进了 HUVECs 的凋亡、炎症和氧化应激，部分原因是通过上调 circ_0004104。微小 RNA-328-3p（miR-328-3p）被证实是 circ_0004104 的靶标。miR-328-3p 干扰在很大程度上逆转了 ox-LDL 暴露下 circ_0004104 沉默介导的 HUVECs 效应。miR-328-3p 与三部分基序 14 的 3'非翻译区相互作用，circ_0004104 通过海绵 miR-328-3p 正向调节 TRIM14 表达。TRIM14 过表达在很大程度上推翻了 ox-LDL 暴露下 miR-328-3p 积累诱导的 HUVECs 影响。
circ_0004104 敲低通过 miR-328-3p 介导的 TRIM14 调节减轻了 ox-LDL 诱导的 HUVECs 功能障碍。
