Zhao Xianlei, Gu Chenjie, Wang Min, Huang Sicong, Gong Xinghan, Kang Muxue, Zhang Tiesong, Shen Jian, Yang Xiaohang, Xi Yongmei, Pan Jianwei
The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu, 322000, China.
Institute of Genetics, Zhejiang University International School of Medicine, Hangzhou, 310000, China.
Mol Neurobiol. 2025 May 22. doi: 10.1007/s12035-025-04952-z.
The prevention and treatment of acute ischemic stroke have been longstanding challenges. Therapies targeting angiogenesis hold promising potential for ischemic injury repair. Reactive oxygen species (ROS) production, induced by the overexpression of NADPH oxidase 4 (NOX4), is a key factor that inhibits angiogenesis during the acute phase of ischemia/reperfusion. Therefore, targeting NOX4 gene expression can reduce ROS production and promote angiogenesis. In this study, human neural progenitor cell (hNPC)-derived-exosomal miRNAs were identified using high-throughput sequencing and online database, with miR-100-5p showing potential to suppress NOX4 expression. We then incorporated exogenous miR-100-5p into hNPC-derived exosomes through electroporation. These miR-100-5p-loaded exosomes were then applied to in vitro models of mouse brain microvascular endothelial cells (bEND.3) subjected to oxygen glucose deprivation and reperfusion (OGD/R). The results showed that miR-100-5p could significantly reduce NOX4 expression and ROS levels induced by OGD/R in bEND.3 cells. Similarly, in vivo analysis showed that mice models of middle cerebral artery occlusion (MCAO) injected with hNPC-derived exosomes loaded with miR-100-5p exhibited smaller brain infarct size, reduced apoptosis, and improved neurological performance compared to MCAO mice receiving PBS. Our findings demonstrate the successful delivery of miR-100-5p via hNPC-derived exosomes and its protective effect on brain microvascular endothelial cells following ischemia injury.
急性缺血性中风的预防和治疗一直是长期存在的挑战。针对血管生成的疗法在缺血性损伤修复方面具有广阔的潜力。由NADPH氧化酶4（NOX4）过表达诱导产生的活性氧（ROS）是缺血/再灌注急性期抑制血管生成的关键因素。因此，靶向NOX4基因表达可以减少ROS产生并促进血管生成。在本研究中，通过高通量测序和在线数据库鉴定了人神经祖细胞（hNPC）衍生的外泌体miRNA，其中miR-100-5p显示出抑制NOX4表达的潜力。然后，我们通过电穿孔将外源性miR-100-5p掺入hNPC衍生的外泌体中。接着将这些装载了miR-100-5p的外泌体应用于经历氧糖剥夺和再灌注（OGD/R）的小鼠脑微血管内皮细胞（bEND.3）的体外模型。结果表明，miR-100-5p可以显著降低OGD/R诱导的bEND.3细胞中NOX4的表达和ROS水平。同样，体内分析表明，与接受PBS的MCAO小鼠相比，注射了装载miR-100-5p的hNPC衍生外泌体的大脑中动脉闭塞（MCAO）小鼠模型的脑梗死面积更小，细胞凋亡减少，神经功能得到改善。我们的研究结果证明了通过hNPC衍生的外泌体成功递送miR-100-5p及其对缺血性损伤后脑微血管内皮细胞的保护作用。
