Department of Neurology, Fujian Medical University Union Hospital, Fuzhou, China.
Department of Rehabilitation, Fujian Medical University Union Hospital, Fuzhou, China.
J Transl Med. 2023 May 3;21(1):297. doi: 10.1186/s12967-023-04125-3.
Available literature documents that ischemic stroke can disrupt the morphology and function of mitochondria and that the latter in other disease models can be preserved by neuropilin-1 (NRP-1) via oxidative stress suppression. However, whether NRP-1 can repair mitochondrial structure and promote functional recovery after cerebral ischemia is still unknown. This study tackled this very issue and explored the underlying mechanism.
Adeno-associated viral (AAV)-NRP-1 was stereotaxically inoculated into the cortex and ipsilateral striatum posterior of adult male Sprague-Dawley (SD) rats before a 90-min transient middle cerebral artery occlusion (tMCAO) and subsequent reperfusion. Lentivirus (LV)-NRP-1 was transfected into rat primary cortical neuronal cultures before a 2-h oxygen-glucose deprivation and reoxygenation (OGD/R) injury to neurons. The expression and function of NRP-1 and its specific protective mechanism were investigated by Western Blot, immunofluorescence staining, flow cytometry, magnetic resonance imaging, transmission electron microscopy, etc. The binding was detected by molecular docking and molecular dynamics simulation.
Both in vitro and in vivo models of cerebral ischemia/reperfusion (I/R) injury presented a sharp increase in NRP-1 expression. The expression of AAV-NRP-1 markedly ameliorated the cerebral I/R-induced damage to the motor function and restored the mitochondrial morphology. The expression of LV-NRP-1 alleviated mitochondrial oxidative stress and bioenergetic deficits. AAV-NRP-1 and LV-NRP-1 treatments increased the wingless integration (Wnt)-associated signals and β-catenin nuclear localization. The protective effects of NRP-1 were reversed by the administration of XAV-939.
NRP-1 can produce neuroprotective effects against I/R injury to the brain by activating the Wnt/β-catenin signaling pathway and promoting mitochondrial structural repair and functional recovery, which may serve as a promising candidate target in treating ischemic stroke.
现有文献资料表明，缺血性脑卒中可破坏线粒体的形态和功能，而在其他疾病模型中，神经纤毛蛋白-1（NRP-1）可通过抑制氧化应激来保护线粒体。然而，NRP-1 是否能修复脑缺血后线粒体的结构并促进功能恢复仍不清楚。本研究解决了这一问题，并探讨了其潜在机制。
在成年雄性 Sprague-Dawley（SD）大鼠大脑中动脉闭塞 90 分钟（tMCAO）及随后的再灌注前，立体定向接种腺相关病毒（AAV）-NRP-1 到大脑皮质和对侧纹状体后部。在神经元氧葡萄糖剥夺和复氧（OGD/R）损伤前，将慢病毒（LV）-NRP-1 转染到大鼠原代皮质神经元培养物中。通过 Western Blot、免疫荧光染色、流式细胞术、磁共振成像、透射电子显微镜等方法研究 NRP-1 的表达和功能及其特定的保护机制。通过分子对接和分子动力学模拟检测结合。
脑缺血/再灌注（I/R）损伤的体内外模型均表现出 NRP-1 表达的急剧增加。AAV-NRP-1 的表达显著改善了脑 I/R 引起的运动功能损伤，并恢复了线粒体形态。LV-NRP-1 的表达减轻了线粒体氧化应激和生物能量缺陷。AAV-NRP-1 和 LV-NRP-1 治疗增加了 Wnt 相关信号和β-连环蛋白核定位。用 XAV-939 处理可逆转 NRP-1 的保护作用。
NRP-1 通过激活 Wnt/β-连环蛋白信号通路，促进线粒体结构修复和功能恢复，对脑 I/R 损伤产生神经保护作用，可能成为治疗缺血性脑卒中的有前途的候选靶点。
