Withaferin A inhibits ferroptosis and protects against intracerebral hemorrhage

• Published in: Neural regeneration research Vol. 18; no. 6; pp. 1308 - 1315
• Main Authors: Zhou, Zi-Xian, Cui, Qi, Zhang, Ying-Mei, Yang, Jia-Xin, Xiang, Wen-Jing, Tian, Ning, Jiang, Yan-Lin, Chen, Mei-Ling, Yang, Bin, Li, Qing-Hua, Liao, Ru-Jia
• Format: Journal Article
• Language: English
• Published: India Wolters Kluwer India Pvt. Ltd 01.06.2023; Medknow Publications & Media Pvt. Ltd; Guangxi Clinical Research Center for Neurological Diseases,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China; Laboratory of Neuroscience,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China; Guangxi Clinical Research Center for Neurological Diseases,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China%Department of Pharmacology,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China%Department of Neurology,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China%Guangxi Clinical Research Center for Neurological Diseases,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China%Laboratory of Neuroscience,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China; Department of Neurology,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China%Laboratory of Neuroscience,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China; Department of Neurology,Affiliated Hospital of Guilin Medical University,Guilin Medical University,Guilin,Guangxi Zhuang Autonomous Region,China; Wolters Kluwer - Medknow; Wolters Kluwer Medknow Publications
• Subjects: behavior; brain injuries; hemorrhagic stroke; ferroptosis; heme oxygenase-1; neuroprotection; nuclear factor e2-related factor 2; nuclear translocator; oxidative stress; stroke; Ferroptosis; Hemorrhage; Oxidative stress; ferroptosis; brain injuries; nuclear factor E2-related factor 2; neuroprotection; hemorrhagic stroke; heme oxygenase-1; behavior; stroke; nuclear translocator; oxidative stress
• ISSN: 1673-5374; 1876-7958
• DOI: 10.4103/1673-5374.355822

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.