环状 RNA_0003307 促进脑微血管内皮细胞在脑缺血再灌注损伤中的血管生成、侵袭和迁移：miRNA-191-5p/CDK6 通路的潜在参与。

CircRNA_0003307 promoted brain microvascular endothelial cell angiogenesis, invasion, and migration in cerebral ischemia-reperfusion injury: Potential involvement of miRNA-191-5p/CDK6 pathway.

机构信息

Department of Neurology, The Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University, Luzhou 646000, Sichuan, China.

Department of Orthopedics, Luzhou People's Hospital, Luzhou 646000, Sichuan, China.

出版信息

Neuroscience. 2024 Nov 12;560:77-89. doi: 10.1016/j.neuroscience.2024.09.025. Epub 2024 Sep 14.

DOI:10.1016/j.neuroscience.2024.09.025

PMID:39284436

Abstract

BACKGROUNDS

The role of miR-191-5p in cerebral ischemia-reperfusion (I/R) injury has been established, with its expression in endothelial cells demonstrating anti-angiogenic effects. A potential circular RNA, circRNA_0003307, has been identified through bioinformatics analysis as a candidate for interaction with miR-191-5p, yet its functional significance in brain I/R injury remains unexplored. We aimed to investigate whether circRNA_0003307 regulates brain microvascular endothelial cell (BMEC) vascular tube formation, invasion, and migration by regulating the miR-191-5p cascade.

METHODS

Mouse BMECs (bEnd.3) were culturedand exposed to oxygen-glucose deprivation (OGD). The effects of circRNA_0003307 on vessel-like tube formation and cellular migration were examined. In addition, we investigated the protective effects of circRNA_0003307 on I/R injury in mice.

RESULTS

The results showed the level of circRNA_0003307 was concentration-dependently increased in OGD-induced bEnd.3 cells. ODG-induction enhanced angiogenesis, migration, and invasion of bEnd.3 cells, which were further promoted by the transfection of pcDNA-0003307. Silencing circRNA_0003307 expression showed the opposite results. The dual luciferase assay demonstrated miRNA-191-5p interacted with circRNA_00033073' UTR, and miRNA-191-5p could bind with CDK6. Meanwhile, circRNA_0003307 promoted the expression of CDK6 by sponging miRNA-191-5p. The overexpression of circRNA_0003307 activated the angiogenesis, migration, and invasion of OGD-induced bEnd.3 cells, which were hindered by miRNA-191-5p mimic or siRNA-CDK6. Thus, circRNA_0003307 promoted ODG-induced angiogenesis, migration, and invasion of bEnd.3 cells by targeting miR-191-5p/CDK6 axis. In vivo, circRNA_0003307 had protective effects on brain I/R injury, including neuroprotection, anti-apoptosis and angiogenesis.

CONCLUSION

CircRNA_0003307 may be a promisingtherapeutictarget forthe treatment of cerebral I/R injury.

摘要

背景

miR-191-5p 在脑缺血再灌注（I/R）损伤中的作用已得到确立，其在内皮细胞中的表达表现出抗血管生成作用。通过生物信息学分析鉴定出一种潜在的环状 RNA，circRNA_0003307，作为与 miR-191-5p 相互作用的候选物，但它在脑 I/R 损伤中的功能意义仍未得到探索。我们旨在研究 circRNA_0003307 是否通过调节 miR-191-5p 级联来调节脑微血管内皮细胞（BMEC）血管管形成、侵袭和迁移。

方法

培养小鼠 BMEC（bEnd.3）并进行氧葡萄糖剥夺（OGD）。检测 circRNA_0003307 对血管样管形成和细胞迁移的影响。此外，我们还研究了 circRNA_0003307 对小鼠 I/R 损伤的保护作用。

结果

结果显示，OGD 诱导的 bEnd.3 细胞中 circRNA_0003307 的水平呈浓度依赖性增加。OGD 诱导增强了 bEnd.3 细胞的血管生成、迁移和侵袭，而过表达 pcDNA-0003307 进一步促进了这些作用。沉默 circRNA_0003307 的表达则呈现相反的结果。双荧光素酶报告基因实验表明 miRNA-191-5p 与 circRNA_0003307 的 3'UTR 相互作用，miRNA-191-5p 可以与 CDK6 结合。同时，circRNA_0003307 通过海绵吸附 miR-191-5p 促进 CDK6 的表达。过表达 circRNA_0003307 可激活 OGD 诱导的 bEnd.3 细胞的血管生成、迁移和侵袭，而 miR-191-5p 模拟物或 siRNA-CDK6 则可抑制这些作用。因此，circRNA_0003307 通过靶向 miR-191-5p/CDK6 轴促进 ODG 诱导的 bEnd.3 细胞的血管生成、迁移和侵袭。在体内，circRNA_0003307 对脑 I/R 损伤具有保护作用，包括神经保护、抗细胞凋亡和血管生成。