Hsa_circ_0005548 knockdown repairs OGD/R-induced damage in human brain microvascular endothelial cells via miR-362-3p/ETS1 axis.

IF 1.7 4区医学 Q4 NEUROSCIENCES

International Journal of Neuroscience Pub Date : 2024-10-01 Epub Date: 2023-09-01 DOI:10.1080/00207454.2023.2246100

Chunlei Chen, Jiguo Xu, Tianrun Huang, Zhuolei Qian

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引用次数: 0

Abstract

Background: Ischemic stroke (IS) is a highly prevalent type of stroke with very high rates of disability and death. As the regulatory role of circular RNAs (circRNAs) in various diseases has been revealed, we constructed a stroke cell model to analyze the action mechanism of hsa_circ_0005548 in IS.

Methods: The abundance of hsa_circ_0005548, microRNA-362-3p (miR-362-3p) and E26 transformation specific-1 (ETS-1) were measured by real-time quantitative polymerase chain reaction (RT-qPCR) or western blot. We constructed an IS cell model in vitro by oxygen-glucose deprivation/reperfusion (OGD/R) treatment and analyzed cell proliferation, apoptosis and inflammatory response through the use of Cell Counting Kit-8 (CCK8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry and Enzyme-linked immunosorbent assay (ELISA), respectively. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were employed for the analysis of the relationship between miR-362-3p and hsa_circ_0005548 or ETS1.

Results: The higher abundance of hsa_circ_0005548 and ETS-1 and lower level of miR-362-3p were observed in human brain microvascular endothelial immortalized (HBMEC-IM) cells under OGD/R. Hsa_circ_0005548 downregulation mitigated OGD/R-induced HBMEC-IM cell injury. Mechanistically, hsa_circ_0005548 targeted miR-362-3p. MiR-362-3p knockdown reversed the effect of hsa_circ_0005548 silencing on OGD/R-induced HBMEC-IM cell injury. ETS1 was validated as a direct target of miR-362-3p, and miR-362-3p attenuated OGD/R-induced HBMEC-IM cell injury by ETS1. Moreover, hsa_circ_0005548 modulated ETS1 via miR-362-3p.

Conclusion: Hsa_circ_0005548 knockdown repairs OGD/R-induced HBMEC-IM cell damage via miR-362-3p/ETS1 axis.

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敲除 Hsa_circ_0005548 可通过 miR-362-3p/ETS1 轴修复 OGD/R 诱导的人脑微血管内皮细胞损伤。

背景：缺血性脑卒中（IS）是一种发病率很高的脑卒中类型，致残率和死亡率都很高。由于循环 RNA（circRNA）在多种疾病中的调控作用已被发现，我们构建了一个中风细胞模型来分析 hsa_circ_0005548 在 IS 中的作用机制：方法：通过实时定量聚合酶链式反应（RT-qPCR）或Western印迹检测hsa_circ_0005548、microRNA-362-3p（miR-362-3p）和E26转化特异性-1（ETS-1）的丰度。我们通过氧-葡萄糖剥夺/再灌注（OGD/R）处理在体外构建了一个 IS 细胞模型，并分别使用细胞计数试剂盒-8（CCK8）、5-乙炔基-2'-脱氧尿苷（EdU）、流式细胞术和酶联免疫吸附试验（ELISA）分析了细胞增殖、凋亡和炎症反应。采用双荧光素酶报告和 RNA 免疫沉淀（RIP）测定分析 miR-362-3p 与 hsa_circ_0005548 或 ETS1 的关系：结果：在OGD/R条件下，人脑微血管内皮永生细胞（HBMEC-IM）中hsa_circ_0005548和ETS-1的丰度较高，而miR-362-3p的水平较低。下调 Hsa_circ_0005548 可减轻 OGD/R 诱导的 HBMEC-IM 细胞损伤。从机制上讲，hsa_circ_0005548靶向miR-362-3p。敲除 miR-362-3p 逆转了 hsa_circ_0005548 沉默对 OGD/R 诱导的 HBMEC-IM 细胞损伤的影响。ETS1被证实是miR-362-3p的直接靶标，miR-362-3p通过ETS1减轻了OGD/R诱导的HBMEC-IM细胞损伤。此外，hsa_circ_0005548通过miR-362-3p调节ETS1：结论：敲除 Hsa_circ_0005548 可通过 miR-362-3p/ETS1 轴修复 OGD/R 诱导的 HBMEC-IM 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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来源期刊

International Journal of Neuroscience 医学-神经科学

CiteScore

5.10

自引率

0.00%

发文量

132

审稿时长

2 months

期刊介绍： The International Journal of Neuroscience publishes original research articles, reviews, brief scientific reports, case studies, letters to the editor and book reviews concerned with problems of the nervous system and related clinical studies, epidemiology, neuropathology, medical and surgical treatment options and outcomes, neuropsychology and other topics related to the research and care of persons with neurologic disorders. The focus of the journal is clinical and transitional research. Topics covered include but are not limited to: ALS, ataxia, autism, brain tumors, child neurology, demyelinating diseases, epilepsy, genetics, headache, lysosomal storage disease, mitochondrial dysfunction, movement disorders, multiple sclerosis, myopathy, neurodegenerative diseases, neuromuscular disorders, neuropharmacology, neuropsychiatry, neuropsychology, pain, sleep disorders, stroke, and other areas related to the neurosciences.