Abstract：
Cerebral ischemic stroke is a serious disease with high mortality and disability rates. However, few neuroprotective drugs have been used for ischemic stroke in the clinic. Two main reasons may be responsible for this failure: difficulty in penetrating the blood-brain barrier (BBB) and easily inactivated in the blood circulation. Ferroptosis, a lipid oxidation-related cell death, plays significant roles in cerebral ischemia-reperfusion injury. We utilized RVG29, a peptide derived from Rabies virus glycoprotein, to obtain BBB-targeted lipid nanoparticles (T-LNPs) in order to investigate whether T-LNPs improved the neuroprotective effects of Ferrostatin-1 (Fer1, an inhibitor of ferroptosis) against cerebral ischemic damage. T-LNPs significantly increased BBB penetration following oxygen/glucose deprivation exposure in an in vitro BBB model and enhanced the fluorescence distribution in brain tissues at 6 h post-administration in a cerebral ischemic murine model. Moreover, T-LNPs encapsulated Fer1 (T-LNPs-Fer1) significantly enhanced the inhibitory effects of Fer1 on ferroptosis by maintaining the homeostasis of NADPH oxidase 4 (NOX4) and glutathione peroxidase 4 (GPX4) signals in neuronal cells after cerebral ischemia. T-LNPs-Fer1 significantly suppressed oxidative stress [heme oxygenase-1 expression and malondialdehyde (the product of lipid ROS reaction)] in neurons and alleviated ischemia-induced neuronal cell death, compared to Fer1 alone without encapsulation. Furthermore, T-LNPs-Fer1 significantly reduced cerebral infarction and improved behavior functions compared to Fer1-treated cerebral ischemic mice after 45-min ischemia/24-h reperfusion. These findings showed that the T-LNPs helped Fer1 penetrate the BBB and improved the neuroprotection of Fer1 against cerebral ischemic damage in experimental stroke, providing a feasible translational strategy for the development of clinical drugs for the treatment of ischemic stroke.
摘要：
缺血性脑卒中是一种死亡率和致残率较高的严重疾病。然而,临床上很少有神经保护药物用于缺血性卒中。造成这种失败的主要原因可能有两个：难以穿透血脑屏障（BBB）以及在血液循环中容易失活。Ferroptosis,脂质氧化相关的细胞死亡，在脑缺血再灌注损伤中起重要作用。我们利用RVG29，一种来自狂犬病病毒糖蛋白的肽，获得BBB靶向的脂质纳米粒（T-LNPs），以研究T-LNPs是否改善了Ferrostatin-1（Fer1，铁凋亡的抑制剂）对脑缺血损伤的神经保护作用。T-LNP在体外BBB模型中暴露于氧/葡萄糖剥夺后显着增加了BBB的渗透，并在脑缺血鼠模型中给药后6小时增强了脑组织中的荧光分布。此外，T-LNPs包裹的Fer1（T-LNPs-Fer1）通过维持脑缺血后神经元细胞中NADPH氧化酶4（NOX4）和谷胱甘肽过氧化物酶4（GPX4）信号的稳态，显着增强了Fer1对铁凋亡的抑制作用。T-LNPs-Fer1显著抑制神经元的氧化应激[血红素加氧酶-1表达和丙二醛（脂质ROS反应的产物）]，减轻缺血诱导的神经元细胞死亡，与未封装的单独Fer1相比。此外，在45分钟缺血/24小时再灌注后，与Fer1处理的脑缺血小鼠相比，T-LNP-Fer1显着减少了脑梗死并改善了行为功能。这些发现表明，T-LNPs有助于Fer1穿透BBB并改善Fer1对实验性中风中脑缺血损伤的神经保护作用，为临床治疗缺血性卒中药物的开发提供可行的转化策略。
