Song Meng-Lu, Sun Yun-Yun, Yin Hai-Jun, Li Yi, Yang Hua
State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 211198, China.
Acta Pharmacol Sin. 2025 Mar 14. doi: 10.1038/s41401-025-01510-0.
Oxidative damage induced by glutamate triggers neuronal death in cerebral ischemic/reperfusion injury. BTB and CNC homology 1 (BACH1) is a major link between the cellular heme level, the redox state and the transcriptional response. p-Coumaric acid (p-CA) is a natural antioxidant that has been shown to ameliorate ischemic/reperfusion injury. In this study, we investigated whether and how p-CA regulated BACH1 in ischemic/reperfusion injury from the perspective of BACH1 subcellular localization and function. Middle cerebral artery occlusion (MCAO) model was established in male mice. MCAO mice were treated with p-CA (50, 100 mg/kg, ip) twice 5 min after MCAO and 5 h after reperfusion operation, respectively. We showed that p-CA treatment exerted dramatic neuroprotective effects, which were associated with the inhibition of BACH1. In HT22 cells, treatment with p-CA (20 μM) ameliorated OGD/R or glutamate-induced oxidative damage and mitochondrial dysfunction through decreasing the protein level of BACH1, the beneficial effect of p-CA was blocked by BACH1 overexpression. We demonstrated that BACH1 level was markedly elevated in the nucleus of HT22 cells under glutamate stimulation, and transcriptionally regulated NADPH oxidase 4 (NOX4) expression, thus mediating ROS outbreak. p-CA treatment activated the activated Cdc42-associated kinase 1 (ACK1)/protein kinase B (AKT) cascade to facilitate the phosphorylation of BACH1, augmented its interaction with chromosome region maintenance 1 (CRM1), thereby leading to the export of BACH1 from the nucleus and degradation mediated by heme-oxidized IRP2 ubiquitin ligase-1 (HOIL-1). In accord with this, administration of ACK1 inhibitor AIM-100 (20 mg/kg, ip) 5 min after MCAO significantly attenuated the neuroprotective effects of p-CA in MCAO mice. We concluded that ACK1/AKT/BACH1 axis may serve as a promising therapeutic approach for the management of ischemic stroke, thereby broadening the clinical utility of p-CA.Keywords: ischemic/reperfusion injury; p-Coumaric acid; BACH1; NOX4; ACK1/AKT; AIM-100.
谷氨酸诱导的氧化损伤引发脑缺血/再灌注损伤中的神经元死亡。BTB和CNC同源蛋白1（BACH1）是细胞血红素水平、氧化还原状态与转录反应之间的主要联系。对香豆酸（p-CA）是一种天然抗氧化剂，已被证明可改善缺血/再灌注损伤。在本研究中，我们从BACH1亚细胞定位和功能的角度，研究了p-CA在缺血/再灌注损伤中是否以及如何调节BACH1。在雄性小鼠中建立大脑中动脉闭塞（MCAO）模型。分别在MCAO后5分钟和再灌注手术后5小时，对MCAO小鼠腹腔注射p-CA（50、100mg/kg）两次。我们发现p-CA治疗具有显著的神经保护作用，这与抑制BACH1有关。在HT22细胞中，用p-CA（20μM）处理可通过降低BACH1蛋白水平改善氧糖剥夺/再灌注（OGD/R）或谷氨酸诱导的氧化损伤和线粒体功能障碍，BACH1过表达可阻断p-CA的有益作用。我们证明，在谷氨酸刺激下，HT22细胞核中BACH1水平显著升高，并转录调节NADPH氧化酶4（NOX4）的表达，从而介导活性氧爆发。p-CA治疗激活活化的Cdc42相关激酶1（ACK1）/蛋白激酶B（AKT）级联反应，促进BACH1的磷酸化，增强其与染色体区域维持蛋白1（CRM1）的相互作用，从而导致BACH1从细胞核输出并由血红素氧化的铁调节蛋白2泛素连接酶-1（HOIL-1）介导降解。与此一致的是，在MCAO后5分钟腹腔注射ACK1抑制剂AIM-100（20mg/kg）可显著减弱p-CA对MCAO小鼠的神经保护作用。我们得出结论，ACK1/AKT/BACH1轴可能是治疗缺血性中风的一种有前景的治疗方法，从而拓宽了p-CA的临床应用范围。关键词：缺血/再灌注损伤；对香豆酸；BACH1；NOX4；ACK1/AKT；AIM-100
