环状RNA circ_0029589作为miR-1197的“海绵”，通过调控RAB22A促进ox- ldl诱导的内皮细胞损伤。

IF 2.1 4区医学 Q3 HEMATOLOGY

Clinical hemorheology and microcirculation Pub Date : 2023-01-01 DOI:10.3233/CH-221657

Dequan He, Zhiliang Li, Youquan Chen, Ming Huang

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引用次数: 3

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)发病的重要因素。此外，环状RNA (circRNA) circ_0029589已被发现参与氧化低密度脂蛋白(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明确。方法:采用ox-LDL处理的人脐静脉内皮细胞(HUVECs)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、microRNA-1197 (miR-1197)和ras相关蛋白RAB22A (RAB22A)的表达。采用3-(4,5 -二甲基-2-噻唑基)- 2,5 -二苯基-2- h -溴化四唑(MTT)、5-乙基-2'-脱氧尿苷(EdU)、流式细胞术、成管和transwell检测细胞增殖、凋亡、血管生成和侵袭。Western blot分析Cleaved-caspase-3、b细胞淋巴瘤-2相关X蛋白(Bax)和RAB22A。采用ELISA试剂盒检测IL-6、IL-1β和肿瘤坏死因子α (TNF-α)水平。采用专用试剂盒检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)水平。生物信息学软件预测了miR-1197与circ_0029589或RAB22A之间的结合，并通过双荧光素酶报告基因和RNA下拉实验证明了这一点。结果:在ox- ldl处理的huvec中，Circ_0029589和RAB22A表达增强，miR-1197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可能通过海绵作用miR-1197影响RAB22A含量。结论:Circ_0090231可能通过miR-1197/RAB22A轴对ox- ldl介导的HUVEC损伤具有保护作用，为AS内皮细胞损伤提供了一种治疗策略。

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

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.

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来源期刊

Clinical hemorheology and microcirculation 医学-外周血管病

CiteScore

4.30

自引率

33.30%

发文量

170

期刊介绍： Clinical Hemorheology and Microcirculation, a peer-reviewed international scientific journal, serves as an aid to understanding the flow properties of blood and the relationship to normal and abnormal physiology. The rapidly expanding science of hemorheology concerns blood, its components and the blood vessels with which blood interacts. It includes perihemorheology, i.e., the rheology of fluid and structures in the perivascular and interstitial spaces as well as the lymphatic system. The clinical aspects include pathogenesis, symptomatology and diagnostic methods, and the fields of prophylaxis and therapy in all branches of medicine and surgery, pharmacology and drug research. The endeavour of the Editors-in-Chief and publishers of Clinical Hemorheology and Microcirculation is to bring together contributions from those working in various fields related to blood flow all over the world. The editors of Clinical Hemorheology and Microcirculation are from those countries in Europe, Asia, Australia and America where appreciable work in clinical hemorheology and microcirculation is being carried out. Each editor takes responsibility to decide on the acceptance of a manuscript. He is required to have the manuscript appraised by two referees and may be one of them himself. The executive editorial office, to which the manuscripts have been submitted, is responsible for rapid handling of the reviewing process. Clinical Hemorheology and Microcirculation accepts original papers, brief communications, mini-reports and letters to the Editors-in-Chief. Review articles, providing general views and new insights into related subjects, are regularly invited by the Editors-in-Chief. Proceedings of international and national conferences on clinical hemorheology (in original form or as abstracts) complete the range of editorial features.