Wei Min, Liu Yi, Li Dongsheng, Wang Xingdong, Wang Xiaodong, Li Yuping, Yan Zhengcun, Zhang Hengzhu
Department of Neurosurgery, Graduate School of Dalian Medical University, Dalian, China.
Department of Neurosurgery, The Yangzhou School of Clinical Medicine of Dalian Medical University, Yangzhou, China.
IBRO Neurosci Rep. 2024 Aug 6;17:161-176. doi: 10.1016/j.ibneur.2024.08.003. eCollection 2024 Dec.
Following recent research advancements, an increasing level of evidence had been published to indicate that celastrol exerted a therapeutic effect on a range of nervous system diseases. This study therefore aimed to investigate the potential involvement of celastrol on ferroptosis and the blood-brain barrier disruption in intracerebral haemorrhage.
We established a rat intracerebral haemorrhage and adrenal pheochromocytoma cell (PC12) OxyHb models using an ACSL4 overexpression vector. Ferroptosis-related indices were assessed using corresponding assay kits, and immunofluorescence and flow cytometry were used to measure reactive oxygen species (ROS) levels. Additionally, quantitative PCR (qPCR) and western blot analyses were conducted to evaluate the expression of key proteins and elucidate the role of celastrol in intracerebral haemorrhage (ICH).
Celastrol significantly improved neurological function scores, blood-brain barrier integrity, and brain water content in rats with ICH. Moreover, subsequent analysis of ferroptosis-related markers, such as Fe2+, ROS, MDA, and SOD, suggested that celastrol exerted a protective effect against the oxidative damage induced by ferroptosis in ICH rats and cells. Furthermore, Western blotting indicated that celastrol attenuated ferroptosis by modulating the expression levels of key proteins, including acyl-CoA synthetase long-chain family member 4 (ACSL4), glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), and anti-transferrin receptor 1 (TFR1) both in vitro and in vivo. ACSL4 overexpression attenuated the neuroprotective effects of celastrol on ICH in vitro. Molecular docking analysis revealed that celastrol interacted with ACSL4 via the GLU107, GLN109, ASN111, and LYS357 binding sites.
Celastrol exerted antioxidant properties and aids in neurological recovery after stroke by suppressing ACSL4 expression during ferroptosis. As such, this drug represented a promising pharmaceutical candidate for the treatment of ICH.
随着近期研究进展，越来越多的证据表明雷公藤红素对一系列神经系统疾病具有治疗作用。因此，本研究旨在探讨雷公藤红素在脑出血中铁死亡及血脑屏障破坏方面的潜在作用。
我们使用ACSL4过表达载体建立了大鼠脑出血和肾上腺嗜铬细胞瘤细胞（PC12）氧合血红蛋白模型。使用相应的检测试剂盒评估铁死亡相关指标，并通过免疫荧光和流式细胞术测量活性氧（ROS）水平。此外，进行定量PCR（qPCR）和蛋白质印迹分析以评估关键蛋白的表达，并阐明雷公藤红素在脑出血（ICH）中的作用。
雷公藤红素显著改善了ICH大鼠的神经功能评分、血脑屏障完整性和脑含水量。此外，随后对铁死亡相关标志物（如Fe2+、ROS、丙二醛和超氧化物歧化酶）的分析表明，雷公藤红素对ICH大鼠和细胞中铁死亡诱导的氧化损伤具有保护作用。此外，蛋白质印迹表明，雷公藤红素在体外和体内均通过调节关键蛋白的表达水平来减轻铁死亡，这些关键蛋白包括酰基辅酶A合成酶长链家族成员4（ACSL4）、谷胱甘肽过氧化物酶4（GPX4）、铁蛋白重链1（FTH1）和抗转铁蛋白受体1（TFR1）。ACSL4过表达减弱了雷公藤红素在体外对ICH的神经保护作用。分子对接分析表明，雷公藤红素通过GLU107、GLN109、ASN111和LYS357结合位点与ACSL4相互作用。
雷公藤红素具有抗氧化特性，并通过在铁死亡过程中抑制ACSL4表达来促进中风后的神经恢复。因此，这种药物是治疗ICH的有前景的候选药物。
