环状 RNA circ_0029589 通过作为 miR-1197 的海绵体来调节 RAB22A，促进 ox-LDL 诱导的内皮细胞损伤。

Circular RNA circ_0029589 promotes ox-LDL-induced endothelial cell injury through regulating RAB22A by serving as a sponge of miR-1197.

机构信息

Department of Cardiology, Heart Center, Zhujiang Hospital of Southern Medical University, Guangzhou, China.

Department of Cardiology, Heart Center, South China hospital, Health Science Center, Shenzhen University, Shenzhen, China.

出版信息

Clin Hemorheol Microcirc. 2023;83(4):359-376. doi: 10.3233/CH-221657.

DOI:10.3233/CH-221657

PMID:36683504

Abstract

BACKGROUND

Dysfunction of endothelial cells is now considered a vital contributor to the pathogenesis of atherosclerosis (AS). Moreover, circular RNA (circRNA) circ_0029589 has been found to be involved in the regulation of oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell damage. Nevertheless, its molecular mechanism in ox-LDL-triggered endothelial cell injury is poorly defined.

METHODS

Human umbilical vein endothelial cells (HUVECs) treated with ox-LDL were applied as cell models of AS. Circ_0029589, microRNA-1197 (miR-1197), and Ras-related protein Rab-22A (RAB22A) expression were detected using real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation, apoptosis, angiogenesis, and invasion were detected using 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, tube formation, and transwell assays. Western blot analysis of Cleaved-caspase-3, B-cell lymphoma-2 related X protein (Bax), and RAB22A. IL-6, IL-1β, and Tumor necrosis factor α (TNF-α) levels were gauged using ELISA kits. Superoxide Dismutase (SOD) activity and Malondiahyde (MDA) level were assessed using special kits. Bioinformatics software predicted the binding between miR-1197 and circ_0029589 or RAB22A, which was proved using dual-luciferase reporter and RNA pull-down assays.

RESULTS

Circ_0029589 and RAB22A expression were strengthened, and miR-1197 was reduced in ox-LDL-treated HUVECs. Importantly, circ_0029589 silencing ameliorated ox-LDL-triggered HUVEC damage via promoting cell proliferation, tube formation ability, invasion, and repressing cell apoptosis, inflammation, and oxidative stress. Mechanical analysis suggested that circ_0029589 might affect RAB22A content through sponging miR-1197.

CONCLUSION

Circ_0090231 might protect against ox-LDL-mediated HUVEC injury via the miR-1197/RAB22A axis, which provides a therapeutic strategy for endothelial cell damage of AS.

摘要

背景

内皮细胞功能障碍现在被认为是动脉粥样硬化（AS）发病机制的重要贡献者。此外，已经发现环状 RNA（circRNA）circ_0029589 参与了氧化型低密度脂蛋白（ox-LDL）诱导的内皮细胞损伤的调节。然而，其在 ox-LDL 触发的内皮细胞损伤中的分子机制尚不清楚。

方法

用 ox-LDL 处理人脐静脉内皮细胞（HUVEC）作为 AS 的细胞模型。采用实时定量聚合酶链反应（RT-qPCR）检测 circ_0029589、微小 RNA-1197（miR-1197）和 Ras 相关蛋白 Rab-22A（RAB22A）的表达。用 3-（4,5-二甲基-2-噻唑基）-2,5-二苯基-2-H-四唑溴盐（MTT）、5-乙炔基-2'-脱氧尿苷（EdU）、流式细胞术、管形成和 Transwell 测定法检测细胞增殖、凋亡、血管生成和侵袭。用 Western blot 分析Cleaved-caspase-3、B 细胞淋巴瘤-2 相关 X 蛋白（Bax）和 RAB22A。用酶联免疫吸附试验（ELISA）试剂盒测定白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）和肿瘤坏死因子α（TNF-α）水平。用特殊试剂盒评估超氧化物歧化酶（SOD）活性和丙二醛（MDA）水平。生物信息学软件预测 miR-1197 与 circ_0029589 或 RAB22A 的结合，并用双荧光素酶报告和 RNA 下拉测定证实。

结果

在 ox-LDL 处理的 HUVEC 中，circ_0029589 和 RAB22A 的表达增强，miR-1197 的表达降低。重要的是，circ_0029589 沉默通过促进细胞增殖、管形成能力、侵袭和抑制细胞凋亡、炎症和氧化应激来改善 ox-LDL 触发的 HUVEC 损伤。力学分析表明，circ_0029589 可能通过海绵 miR-1197 来影响 RAB22A 的含量。