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.
Neuroscience. 2024 Nov 12;560:77-89. doi: 10.1016/j.neuroscience.2024.09.025. Epub 2024 Sep 14.
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.
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.
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.
CircRNA_0003307 may be a promisingtherapeutictarget forthe treatment of cerebral I/R injury.
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。
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。
circRNA_0003307 可能是治疗脑 I/R 损伤的有前途的治疗靶点。
