目的:据报道,Rasd1与神经毒性相关,新陈代谢,和节奏,但其在蛛网膜下腔出血(SAH)的情况下的效果仍不清楚。脑白质损伤(WMI)和铁性凋亡参与了SAH后早期脑损伤(EBI)的发生。在这项工作中,我们已经研究了Rasd1是否可以引起铁凋亡并有助于SAH诱导的WMI。
方法:用于Rasd1敲低/过表达的慢病毒在SAH诱导前7天通过脑室内(i.c.v)注射施用。SAH等级,脑含水量,短期和长期的神经行为,蛋白质印迹,实时PCR,ELISA,生化估计,免疫荧光,扩散张量成像(DTI),和透射电子显微镜(TEM)进行了系统地。此外,京尼平,选择性解偶联蛋白2(UCP2)抑制剂,用于原代神经元和少突胶质细胞共培养,用于进一步的体外机制研究。
结果:Rasd1敲低改善了神经行为,神经胶质极化,氧化应激,神经炎症,铁性凋亡,和脱髓鞘.相反,Rasd1过表达通过升高活性氧(ROS)的水平加剧了这些变化,炎性细胞因子,MDA,自由铁,和NCOA4,以及有助于降低UCP2,GPX4,铁蛋白,和GSH机械。根据体外研究,Rasd1可通过抑制UCP2,增加活性氧(ROS),并激活NCOA4介导的铁细胞吞噬。
结论:可以得出结论,Rasd1在SAH后的WMI中少突胶质细胞铁蛋白凋亡中起调节作用。
Rasd1 has been reported to be correlated with neurotoxicity, metabolism, and rhythm, but its effect in case of subarachnoid hemorrhage (SAH) remained unclear. White matter injury (WMI) and ferroptosis participate in the early brain injury (EBI) after SAH. In this work, we have investigated whether
Rasd1 can cause ferroptosis and contribute to SAH-induced WMI.
Lentivirus for
Rasd1 knockdown/overexpression was administrated by intracerebroventricular (i.c.v) injection at 7 days before SAH induction. SAH grade, brain water content, short- and long-term neurobehavior, Western blot, real-time PCR, ELISA, biochemical estimation, immunofluorescence, diffusion tensor imaging (DTI), and transmission electron microscopy (TEM) were systematically performed. Additionally, genipin, a selective uncoupling protein 2(UCP2) inhibitor, was used in primary neuron and oligodendrocyte co-cultures for further in vitro mechanistic studies.
Rasd1 knockdown has improved the neurobehavior, glia polarization, oxidative stress, neuroinflammation, ferroptosis, and demyelination. Conversely,
Rasd1 overexpression aggravated these changes by elevating the levels of reactive oxygen species (ROS), inflammatory cytokines, MDA, free iron, and NCOA4, as well as contributing to the decrease of the levels of UCP2, GPX4, ferritin, and GSH mechanistically. According to the in vitro study,
Rasd1 can induce oligodendrocyte ferroptosis through inhibiting UCP2, increasing reactive oxygen species (ROS), and activating NCOA4-mediated ferritinophagy.
It can be concluded that Rasd1 exerts a modulated role in oligodendrocytes ferroptosis in WMI following SAH.