Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Institute of Neurology, Affiliated Hospital of Guangdong Medical University, 57 South Renmin Road, Zhanjiang, 524001, China.
Department of Pharmacology and Toxicology, Boonshoft School of Medicine, Wright State University, Dayton, OH, 45430, USA.
Transl Stroke Res. 2020 Oct;11(5):1148-1164. doi: 10.1007/s12975-020-00794-0. Epub 2020 Apr 14.
The role of miR-503 in brain endothelium and ischemic stroke (IS) remains unclear. We aimed to study the relationship between plasma miR-503 and the onset time, severity, subtypes, and von Willebrand Factor (vWF) level in IS patients and to investigate the roles and underlying mechanisms of miR-503 in middle cerebral artery occlusion (MCAO) mice and cultured cerebral vascular endothelial cells (ECs). In MCAO mice, the effects of plasma from acute severe IS patients (ASS) with or without miR-503 antagomir on brain and ECs damage were determined. In cultured human ECs, the effects of miR-503 overexpression or knockdown on the monolayer permeability, apoptosis, ROS, and NO generation were investigated. For mechanism study, the PI3K/Akt/eNOS pathway, cleaved caspase-3, and bcl-2 were analyzed. Results showed that plasma miR-503 was significantly increased in IS patients, especially in acute period and severe cases and subtypes of LAA and TACI, and was positively correlated with vWF. Logistic analysis indicated that miR-503 was an independent risk factor for IS, with the area under curve of 0.796 in ROC analysis. In MCAO mice, ASS pretreatment aggravated neurological injury, BBB damage, brain edema, CBF reduction, and decreased NO production while increased apoptosis and ROS generation in brain ECs, which were partly abolished by miR-503 antagomir. In cultured ECs, miR-503 overexpression and knockdown confirmed its effects on regulating monolayer permeability, cell apoptosis, NO, and ROS generation via PI3K/Akt/eNOS pathway or bcl-2 and cleaved caspase-3 proteins. These together indicate that miR-503 is a promising biomarker and novel therapeutic target for IS.
miR-503 在脑内皮细胞和缺血性中风 (IS) 中的作用尚不清楚。本研究旨在探讨 IS 患者血浆 miR-503 与发病时间、严重程度、亚型和血管性血友病因子 (vWF) 水平的关系，并研究 miR-503 在大脑中动脉闭塞 (MCAO) 小鼠和培养的脑血管内皮细胞 (EC) 中的作用及其潜在机制。在 MCAO 小鼠中，观察急性重度 IS 患者（ASS）合并或不合并 miR-503 拮抗剂的血浆对脑和 EC 损伤的影响。在培养的人 EC 中，观察 miR-503 过表达或敲低对单层通透性、细胞凋亡、ROS 和 NO 生成的影响。为了研究机制，分析了 PI3K/Akt/eNOS 通路、caspase-3 及 bcl-2。结果显示，IS 患者，尤其是急性期和重度患者、LAA 和 TACI 亚型患者，血浆 miR-503 显著升高，与 vWF 呈正相关。Logistic 分析表明，miR-503 是 IS 的独立危险因素，ROC 分析曲线下面积为 0.796。在 MCAO 小鼠中，ASS 预处理加重神经损伤、BBB 损伤、脑水肿、CBF 减少和 NO 生成减少，同时增加脑 EC 凋亡和 ROS 生成，这些作用部分被 miR-503 拮抗剂所拮抗。在培养的 EC 中，miR-503 过表达和敲低证实其通过 PI3K/Akt/eNOS 通路或 bcl-2 和 cleaved caspase-3 蛋白调节单层通透性、细胞凋亡、NO 和 ROS 生成。这些结果表明，miR-503 是 IS 有前途的生物标志物和新型治疗靶点。
