Department of Neurology, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Autoimmunity. 2023 Dec;56(1):2201405. doi: 10.1080/08916934.2023.2201405.
Stroke is an acute cerebrovascular disease that is now the most important cause of death due to brain problems in our country. CircRNAs are RNA circles that have been extensively involved in the disease. We aimed to investigate the mechanism of circ_0129657 in the pathogenesis of stroke. In this study, quantitative real-time polymerase chain reaction (RT-qPCR) and western blot assays were used to assess the expression of circ_0129657, miR-194-5p, and glia maturation factor beta (GMFB). Cell viability was measured by Cell Counting Kit-8 (CCK-8) assay. 5-Ethynyl-2'-Deoxyuridine (EdU) assay was used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis. Dual-luciferase reporter, RNA pull-down, and RNA immunoprecipitation (RIP) assays were used to assess the relationship between miR-194-5p and circ_0129657 or GMFB. Mouse middle cerebral artery occlusion (MCAO) model was applied to mimic the cerebral ischemia/reperfusion injury. Our data showed that the levels of circ_0129657 and GMFB were significantly increased and the expression of miR-194-5p was significantly decreased in oxygen-glucose deprivation (OGD)-induced human brain microvascular endothelial cells (HBMECs). Silencing circ_0129657 expression in OGD-induced HBMECs could promote cell viability and cell proliferation. Moreover, circ_0129657 depletion also could inhibit apoptosis and inflammatory factor secretion. Circ_0129657 functioned as a sponge for miR-194-5p and could regulate GMFB expression miR-194-5p competition. Furthermore, miR-194-5p downregulation or GMFB restoration could partially reverse the effects of circ_0129657 silencing on cell biological properties in OGD-induced HBMECs. Meanwhile, circ_0129657 knockdown decreased cerebral infarction volume and neurological impairment in MCAO mouse models. In conclusion, our findings suggest that circ_0129657 can inhibit cell proliferation and promote apoptosis and inflammatory factor secretion in HBMECs after oxygen-glucose deprivation miR-194-5p/GMFB axis, providing evidence that circ_0129657 has the potential as a useful biological molecular marker in the diagnosis of stroke.
中风是一种急性脑血管病,是我国目前因脑部问题导致死亡的最重要原因。circRNAs 是一种 RNA 环,它广泛参与了疾病的发生。我们旨在研究 circ_0129657 在中风发病机制中的作用。在这项研究中,使用定量实时聚合酶链反应 (RT-qPCR) 和 Western blot 分析来评估 circ_0129657、miR-194-5p 和神经胶质细胞成熟因子β (GMFB) 的表达。通过 Cell Counting Kit-8 (CCK-8) 测定法测量细胞活力。5-乙炔基-2'-脱氧尿苷 (EdU) 测定法用于检测细胞增殖。流式细胞术用于检测细胞凋亡。双荧光素酶报告、RNA 下拉和 RNA 免疫沉淀 (RIP) 测定法用于评估 miR-194-5p 与 circ_0129657 或 GMFB 之间的关系。应用小鼠大脑中动脉闭塞 (MCAO) 模型模拟脑缺血再灌注损伤。我们的数据显示,在氧葡萄糖剥夺 (OGD) 诱导的人脑血管内皮细胞 (HBMEC) 中,circ_0129657 和 GMFB 的水平显著升高,miR-194-5p 的表达显著降低。在 OGD 诱导的 HBMECs 中沉默 circ_0129657 的表达可以促进细胞活力和细胞增殖。此外,circ_0129657 耗竭还可以抑制细胞凋亡和炎症因子的分泌。Circ_0129657 作为 miR-194-5p 的海绵,可调节 GMFB 表达 miR-194-5p 竞争。此外,miR-194-5p 的下调或 GMFB 的恢复可以部分逆转 OGD 诱导的 HBMECs 中 circ_0129657 沉默对细胞生物学特性的影响。同时,circ_0129657 敲低可减少 MCAO 小鼠模型中的脑梗死体积和神经损伤。总之,我们的研究结果表明,circ_0129657 可抑制 OGD 后 HBMECs 中的细胞增殖,促进细胞凋亡和炎症因子的分泌 miR-194-5p/GMFB 轴,为 circ_0129657 作为中风诊断有用的生物分子标志物提供了证据。