Clin Lab. 2022 Aug 1;68(8). doi: 10.7754/Clin.Lab.2021.210834.
Ischemic stroke refers to ischemic necrosis or softening of limited brain tissue caused by ischemia and hypoxia due to impaired blood circulation in the brain. Ischemic stroke is a major classification of cerebrovascular disease, accounting for about 80% of patients with cerebrovascular disease in China, with a high rate of disability and death. Recently, miRNAs were reported to participate in ischemic stroke pathogenesis and development. In the study, we aimed to study the role and underlying mechanism of miR-641 in ischemic stroke.
Serum samples were extracted from acute ischemic stroke (AIS) patients and healthy controls. The oxygen-glucose deprivation/reoxygenation (OGD/R) method was used to treat SH-SY5Y cells to construct an ischemic stroke in vitro model. Real-time quantitative polymerase chain reaction (qRT-PCR) assay and western blot analysis were conducted to detect miR-641 and MCL-1 expressions. The targeted relationship between miR-641 and MCL-1 was confirmed by dual-luciferase reporter, RNA pull-down, and rescue assays. CCK-8, flow cytometry, and ELISA assays were performed to measure cell viability, apoptosis, and inflammation. The activation of the Wnt/β-catenin pathway was verified by western blot assay.
MiR-641 was increased while MCL-1 was decreased in serum samples from AIS patients, serving as highly-sensitive biomarkers in AIS diagnosis. After OGD/R treatment, SH-SY5Y cell viability, and MCL-1 expression were decreased, along with increased miR-641 expression, cell apoptosis, and inflammation. MiR-641 aggravated while MCL-1 mitigated OGD/R-triggered injury and inflammation in SH-SY5Y cells. MCL-1 was a downstream target of miR-641, which could be negatively regulated by miR-641. Finally, miR-641 exacerbated the progression of OGD/R-triggered SH-SY5Y cell injury via the MCL-1/Wnt/β-catenin pathway.
MiR-641 may be a novel therapeutic agent for ischemic stroke by modulating the MCL-1/Wnt/β-catenin axis on neuronal damage in brain tissue in the ischemic region after ischemic stroke.
缺血性脑卒中是指由于脑血液循环障碍导致的局部脑组织缺血缺氧性坏死或软化。缺血性脑卒中是脑血管病的主要分类，约占中国脑血管病患者的 80%，具有高致残率和高死亡率。最近，有研究报道 miRNA 参与了缺血性脑卒中的发病机制和发展。本研究旨在探讨 miR-641 在缺血性脑卒中中的作用及潜在机制。
从急性缺血性脑卒中（AIS）患者和健康对照者中提取血清样本。采用氧葡萄糖剥夺/再复氧（OGD/R）方法处理 SH-SY5Y 细胞，构建体外缺血性脑卒中模型。采用实时定量聚合酶链反应（qRT-PCR）和 Western blot 分析检测 miR-641 和 MCL-1 的表达。通过双荧光素酶报告、RNA 下拉和拯救实验验证 miR-641 和 MCL-1 之间的靶向关系。采用 CCK-8、流式细胞术和 ELISA 检测细胞活力、细胞凋亡和炎症。Western blot 检测 Wnt/β-catenin 通路的激活情况。
AIS 患者血清样本中 miR-641 升高，MCL-1 降低，可作为 AIS 诊断的高度敏感生物标志物。OGD/R 处理后，SH-SY5Y 细胞活力和 MCL-1 表达降低，同时 miR-641 表达升高，细胞凋亡和炎症增加。miR-641 加重而 MCL-1 减轻 OGD/R 诱导的 SH-SY5Y 细胞损伤和炎症。MCL-1 是 miR-641 的下游靶点，可被 miR-641 负调控。最后，miR-641 通过 MCL-1/Wnt/β-catenin 通路加剧了 OGD/R 诱导的 SH-SY5Y 细胞损伤的进展。
miR-641 可能通过调节缺血性脑卒中后缺血区域脑组织神经元损伤中的 MCL-1/Wnt/β-catenin 轴，成为治疗缺血性脑卒中的一种新的治疗药物。
