Liu Qie, Zhang Lu, Xu Xin
Department of Neurology, Daqing Oilfield General Hospital, Daqing City, Heilongjiang Province, China.
Department of Neurology, Daqing Oilfield General Hospital, Daqing City, Heilongjiang Province, China.
Clinics (Sao Paulo). 2025 Jan 29;80:100581. doi: 10.1016/j.clinsp.2025.100581. eCollection 2025.
The authors explored differentially expressed circRNAs in Acute Ischemic Stroke (AIS) and revealed the role and potential downstream molecular mechanisms of circLOC375190.
circLOC375190 expression was modulated by lentiviral injection in the brain of transient Middle Cerebral Artery Occlusion (tMCAO) mice. Neurological dysfunction was assessed, as well as infarction size, histopathological changes, and neuronal apoptosis in tMCAO mice. An in vitro Oxygen-Glucose Deprivation/Reoxygenation (OGD/R) PC-12 cell model was established. PC-12 cells were transfected and evaluated for viability, cytotoxicity, apoptosis, and autophagy. Inflammatory factors in mouse brain tissues and PC-12 cells were examined via enzyme-linked immunosorbent assay, and related genes were measured via real-time reverse transcriptase-polymerase chain reaction and Western blot. The ring structure of circLOC375190 was assessed by actinomycin-D and RNase-R assays. circRNA targeting to downstream factors was assessed by Fluorescence in situ hybridization assay, dual luciferase reporter assay, and RNA immunoprecipitation assay.
circLOC375190 level was increased in tMCAO mice. Knocking down circLOC375190 reduced infarct size, attenuated cerebral pathological injury and neuronal apoptosis, and inhibited inflammatory damage and autophagy in tMCAO mice. circLOC375190 knockdown enhanced neuronal viability and reduced cytotoxicity, apoptosis, and autophagy in OGD/R-treated PC12 cells. Mechanistically, circLOC375190 acted as a sponge for miR-93-5p to upregulate MAP kinase interacting serine/threonine kinase 2 expression and activate the mechanistic target of rapamycin complex 1/transcription factor EB pathway.
circLOC375190 exacerbates tMCAO-mediated neurological injury by regulating neuronal autophagy.
作者探究急性缺血性脑卒中（AIS）中差异表达的环状RNA（circRNA），并揭示circLOC375190的作用及潜在的下游分子机制。
通过慢病毒注射调节短暂性大脑中动脉闭塞（tMCAO）小鼠脑内circLOC375190的表达。评估tMCAO小鼠的神经功能障碍、梗死面积、组织病理学变化和神经元凋亡。建立体外氧糖剥夺/复氧（OGD/R）PC-12细胞模型。对PC-12细胞进行转染，并评估其活力、细胞毒性、凋亡和自噬情况。通过酶联免疫吸附测定法检测小鼠脑组织和PC-12细胞中的炎症因子，并通过实时逆转录聚合酶链反应和蛋白质免疫印迹法检测相关基因。通过放线菌素-D和核糖核酸酶-R试验评估circLOC375190的环状结构。通过荧光原位杂交试验、双荧光素酶报告基因试验和RNA免疫沉淀试验评估靶向下游因子的circRNA。
tMCAO小鼠中circLOC375190水平升高。敲低circLOC375190可减小梗死面积，减轻脑病理损伤和神经元凋亡，并抑制tMCAO小鼠的炎症损伤和自噬。敲低circLOC375190可增强OGD/R处理的PC12细胞的神经元活力，降低细胞毒性、凋亡和自噬。机制上，circLOC375190作为miR-93-5p的海绵，上调丝裂原活化蛋白激酶相互作用丝氨酸/苏氨酸激酶2的表达，并激活雷帕霉素靶蛋白复合物1/转录因子EB途径。
circLOC375190通过调节神经元自噬加重tMCAO介导的神经损伤。
