Table_1_TAK1 inhibition mitigates intracerebral hemorrhage-induced brain injury through reduction of oxidative stress and neuronal pyroptosis via the NRF2 signaling pathway.docx

IntroductionIntracerebral hemorrhage (ICH) often triggers oxidative stress through reactive oxygen species (ROS). Transforming growth factor-β-activated kinase 1 (TAK1) plays a pivotal role in regulating oxidative stress and inflammation across various diseases. 5Z-7-Oxozeaenol (OZ), a specific inhibitor of TAK1, has exhibited therapeutic effects in various conditions. However, the impact of OZ following ICH and its underlying molecular mechanisms remain elusive. This study aimed to explore the possible role of OZ in ICH and its underlying mechanisms by inhibiting oxidative stress-mediated pyroptosis.
MethodsAdult male Sprague-Dawley rats were subjected to an ICH model, followed by treatment with OZ. Neurobehavioral function, blood-brain barrier integrity, neuronal pyroptosis, and oxidative stress markers were assessed using various techniques including behavioral tests, immunofluorescence staining, western blotting, transmission electron microscopy, and biochemical assays.
ResultsOur study revealed that OZ administration significantly inhibited phosphorylated TAK1 expression post-ICH. Furthermore, TAK1 blockade by OZ attenuated blood-brain barrier (BBB) disruption, neuroinflammation, and oxidative damage while enhancing neurobehavioral function. Mechanistically, OZ administration markedly reduced ROS production and oxidative stress by facilitating nuclear factor-erythroid 2-related factor 2 (NRF2) nuclear translocation. This was accompanied by a subsequent suppression of the NOD-like receptor protein 3 (NLRP3) activation-mediated inflammatory cascade and neuronal pyroptosis.
DiscussionOur findings highlight that OZ alleviates brain injury and oxidative stress-mediated pyroptosis via the NRF2 pathway. Inhibition of TAK1 emerges as a promising approach for managing ICH.

引言
脑出血（Intracerebral hemorrhage, ICH）常通过活性氧（reactive oxygen species, ROS）诱发氧化应激。转化生长因子-β激活激酶1（Transforming growth factor-β-activated kinase 1, TAK1）在多种疾病的氧化应激与炎症调控过程中发挥关键作用。5Z-7-氧代土曲霉醇（5Z-7-Oxozeaenol, OZ）作为TAK1的特异性抑制剂，已在多种疾病模型中展现出治疗潜力。然而，OZ对脑出血后的干预效果及其潜在分子机制仍不明晰。本研究旨在通过探究OZ抑制氧化应激介导的细胞焦亡这一途径，明确其在脑出血中的潜在作用与分子机制。

方法
本研究采用成年雄性斯普拉格-道利（Sprague-Dawley）大鼠构建脑出血模型，随后给予OZ干预。通过行为学测试、免疫荧光染色、蛋白免疫印迹（western blotting）、透射电子显微镜及生化检测等多种技术手段，评估大鼠的神经行为功能、血脑屏障完整性、神经元焦亡情况及氧化应激标志物水平。

结果
本研究发现，脑出血后给予OZ可显著抑制磷酸化TAK1的表达。进一步研究显示，OZ对TAK1的阻断作用可减轻血脑屏障（blood-brain barrier, BBB）破坏、神经炎症与氧化损伤，同时改善神经行为功能。机制层面，OZ可通过促进核因子E2相关因子2（nuclear factor-erythroid 2-related factor 2, NRF2）的核转位，显著减少活性氧生成与氧化应激；伴随这一过程的是NOD样受体蛋白3（NOD-like receptor protein 3, NLRP3）激活介导的炎症级联反应与神经元焦亡被显著抑制。

讨论
本研究结果表明，OZ可通过NRF2通路减轻脑损伤与氧化应激介导的细胞焦亡。综上，抑制TAK1有望成为治疗脑出血的潜在有效策略。