中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (6): 1308-1315.doi: 10.4103/1673-5374.355822

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

醉茄素A干预可阻断铁死亡减轻脑出血损伤

  

  • 出版日期:2023-06-15 发布日期:2023-01-05
  • 基金资助:
    广西珠江自治区自然科学基金（2020GXNSFA259036）；广西科技项目（GuikeAD17129015）；广西神经损伤修复项目（GuikeZY21195042）；广西研究生教育创新项目（YCW2021246，YCW202225）

Withaferin A inhibits ferroptosis and protects against intracerebral hemorrhage

Zi-Xian Zhou1, 2, #, Qi Cui1, 2, #, Ying-Mei Zhang1, 2, Jia-Xin Yang1, 2, Wen-Jing Xiang1, 2, Ning Tian1, 3, Yan-Lin Jiang4, Mei-Ling Chen2, Bin Yang3, Qing-Hua Li1, 2, 3, Ru-Jia Liao1, 2, 3, *#br#   

  1. 1Laboratory of Neuroscience, Affiliated Hospital of Guilin Medical University, Guilin Medical University, Guilin, Guangxi Zhuang Autonomous Region, China; 2Department of Neurology, Affiliated Hospital of Guilin Medical University, Guilin Medical University, Guilin, Guangxi Zhuang Autonomous Region, China; 3Guangxi Clinical Research Center for Neurological Diseases, Affiliated Hospital of Guilin Medical University, Guilin Medical University, Guilin, Guangxi Zhuang Autonomous Region, China; 4Department of Pharmacology, Affiliated Hospital of Guilin Medical University, Guilin Medical University, Guilin, Guangxi Zhuang Autonomous Region, China
  • Online:2023-06-15 Published:2023-01-05
  • Contact: Ru-Jia Liao, PhD, liaorujia@hotmail.com.
  • Supported by:
    This work was supported by the Natural Science Foundation of Guangxi Zhuang Autonomous Region, No. 2020GXNSFAA259036 (to RJL); the Guangxi Science and Technology Project, No. Guike AD17129015 (to QHL); Guangxi Research and Innovation Base for Basic and Clinical Application of Nerve Injury and Repair Project, No. Guike ZY21195042 (to QHL); the Innovation Projects of Guangxi Graduate Education, Nos. YCSW2021246 (to ZXZ), YCSW2021254 (to WJX).

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摘要：

近期研究发现抑制铁死亡可显著改善脑出血预后，有报道醉茄素A可阻断脑出血后铁死亡信号通路，但调控机制尚不清楚。实验探讨了醉茄素A通过调控氧化应激和铁死亡发挥脑出血后的神经保护作用及机制。①采用自体尾动脉血注入尾状核诱导小鼠脑出血模型，并使用血红素诱导细胞损伤构建脑出血体外模型。②小鼠脑出血术后立即开始每日脑室内注射一次醉茄素A（每只0.1、1 或 5 μg/kg），持续7 d后，发现醉茄素A治疗可显著减轻小鼠脑出血7 d后脑组织损伤、铁沉积，并以剂量依赖性方式改善神经功能障碍。③在体外实验中通过细胞活力检测发现，醉茄素A可保护 SH-SY5Y 神经元细胞免受血红素诱导的细胞损伤。此外在体内和体外研究中，ELISA检测结果均显示醉茄素A治疗可显著降低脑出血后氧化应激标志物丙二醛的水平，并提高抗氧化应激标志物超氧化物歧化酶和谷胱甘肽过氧化物酶。④蛋白质印迹、qPCR和免疫荧光染色检测结果表明，在醉茄素A治疗下，醉茄素A激活了核因子E2相关因子2（Nrf2）/血红素加氧酶1（HO-1）信号轴，并促进了Nrf2从细胞质转移至细胞核并上调HO-1的表达，而在沉默Nrf2表达后完全逆转了醉茄素A处理诱导的HO-1表达、氧化应激和保护作用。⑤进一步检测了铁死亡标记蛋白、脂质过氧化物的终产物和脂质活性氧的水平，显示SH-SY5Y神经元细胞系在暴露于血红素后，用 醉茄素A处理可显著改善细胞铁死亡，而同时给予HO-1抑制剂后，醉茄素A对血红素诱导的SH-SY5Y神经元细胞系铁死亡的神经保护作用减弱。⑥给予细胞铁死亡拮抗剂ferrostatin-1发现， 醉茄素A与ferrostatin-1的组合改善了血红素诱导的 SH-SY5Y神经元细胞损伤，抑制细胞铁死亡。⑦上述数据显示，醉茄素A通过激活Nrf2/HO-1通路减轻脑出血损伤引起的铁死亡和氧化应激，由此推断醉茄素A可能是脑出血治疗的潜在治疗药物。

https://orcid.org/0000-0001-6928-4003 (Ru-Jia Liao)

关键词: 行为, 脑损伤, 脑出血, 铁死亡, 血红素Oxygenase-1, 神经保护, 核因子E2-Related因子2, 核转运体, 氧化应激, 脑卒中

Abstract: Recent studies have indicated that suppressing oxidative stress and ferroptosis can considerably improve the prognosis of intracerebral hemorrhage (ICH). Withaferin A (WFA), a natural compound, exhibits a positive effect on a number of neurological diseases. However, the effects of WFA on oxidative stress and ferroptosis-mediated signaling pathways to ICH remain unknown. In this study, we investigated the neuroprotective effects and underlying mechanism for WFA in the regulation of ICH-induced oxidative stress and ferroptosis. We established a mouse model of ICH by injection of autologous tail artery blood into the caudate nucleus and an in vitro cell model of hemin-induced ICH. WFA was injected intracerebroventricularly at 0.1, 1 or 5 μg/kg once daily for 7 days, starting immediately after ICH operation. WFA markedly reduced brain tissue injury and iron deposition and improved neurological function in a dose-dependent manner 7 days after cerebral hemorrhage. Through in vitro experiments, cell viability test showed that WFA protected SH-SY5Y neuronal cells against hemin-induced cell injury. Enzyme-linked immunosorbent assays in vitro and in vivo showed that WFA markedly decreased the level of malondialdehyde, an oxidative stress marker, and increased the activities of anti-oxidative stress markers superoxide dismutase and glutathione peroxidase after ICH. Western blot assay, quantitative polymerase chain reaction and immunofluorescence results demonstrated that WFA activated the nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling axis, promoted translocation of Nrf2 from the cytoplasm to nucleus, and increased HO-1 expression. Silencing Nrf2 with siRNA completely reversed HO-1 expression, oxidative stress and protective effects of WFA. Furthermore, WFA reduced hemin-induced ferroptosis. However, after treatment with an HO-1 inhibitor, the neuroprotective effects of WFA against hemin-induced ferroptosis were weakened. MTT test results showed that WFA combined with ferrostatin-1 reduced hemin-induced SH-SY5Y neuronal cell injury. Our findings reveal that WFA treatment alleviated ICH injury-induced ferroptosis and oxidative stress through activating the Nrf2/HO-1 pathway, which may highlight a potential role of WFA for the treatment of ICH. 

Key words: behavior, brain injuries, hemorrhagic stroke, ferroptosis, heme oxygenase-1, neuroprotection, nuclear factor E2-related factor 2, nuclear translocator, oxidative stress, stroke