针灸和草药（英文）2026，Vol.6Issue(1)：73-90,18.DOI:10.1097/HM9.0000000000000182
柽柳素靶向和促进Nrf2表达抑制缺血性脑卒中大鼠神经元铁死亡
Tamarixetin suppresses neuronal ferroptosis in ischemic stroke rats by targeting and facilitating nuclear factor erythroid-2-related factor 2 expression
摘要
Abstract
Objective:Neuronal ferroptosis has emerged as a promising therapeutic target for ischemic stroke.Tamarixetin,a natural dietary flavonoid,exerts protective effects against ischemic stroke by modulating neuroinflammatory responses and mitigating oxidative stress.However,its potential role in regulating neuronal ferroptosis remains unclear. Methods:A rat model of middle cerebral artery occlusion and reperfusion and an erastin-treated SH-SY5Y cell model were used for in vivo and in vitro experiments,respectively.The neurological function of the rats was evaluated using a series of behavioral tests,the Garcia scoring system,and 2,3,5-triphenyltetrazolium chloride staining.Neuronal damage was detected via immunofluorescence staining and terminal deoxynucleotidyl transferase(TdT)-mediated deoxyuridine triphosphate(dUTP)nick-end labeling.Commercial kits,western blotting,and coimmunoprecipitation were used to analyze neuronal ferroptosis and the activation of the Nuclear factor erythroid-2-related factor 2(Nrf2)signaling pathway.The direct target protein of tamarixetin was examined using the cellular thermal shift assay,drug affinity-responsive target stability assay,surface plasmon resonance,and molecular docking.Cellular Nrf2 was knocked down using small interfering RNA. Results:Tamarixetin mitigated the neurological dysfunctions of middle cerebral artery occlusion and reperfusion(MCAO/R)rats,including motor dysfunction,limb coordination impairment,neurological deficit,cerebral infarction,and reduced neuronal loss.Furthermore,it alleviated neuronal ferroptosis in vivo and in vitro by lowering the levels of iron ions,reactive oxygen species,malondialdehyde,and acyl-CoA synthetase long-chain family member 4 and upregulating the expression of superoxide dismutase,glutathione,glutathione peroxidase 4,heme oxygenase-1,and solute carrier family 7 member 11.Tamarixetin activated the Nrf2 signaling pathway by suppressing Keap1 protein expression,weakening the interaction between Keap1 and Nrf2,upregulating Nrf2 protein expression and nuclear translocation,and promoting antioxidant response element activity.Nrf2 is the direct binding protein of tamarixetin.It specifically interacts with amino acid residues at arginine 72,arginine 515,and lysine 518.The effects of tamarixetin on Nrf2 signaling pathway activation and neuronal ferroptosis inhibition were abrogated in Nrf2 knockdown cells challenged with erastin. Conclusions:Our findings not only identify tamarixetin as a novel ferroptosis inhibitor but also elucidate its mechanism of action via direct binding and Nrf2 pathway activation,providing a promising therapeutic candidate for ischemic stroke.关键词
铁死亡/缺血性脑卒中/柽柳素/神经元/核因子红系2相关因子2Key words
Ferroptosis/Ischemic stroke/Tamarixetin/Neuron/Nuclear factor erythroid-2-related factor 2引用本文复制引用
杨艳秋,巩仔鹏,侯悦,方铭霞,孟庆琦,米妍,徐丽彬,郭华,刘月阳,李明忠,Nanik Siti Aminah..柽柳素靶向和促进Nrf2表达抑制缺血性脑卒中大鼠神经元铁死亡[J].针灸和草药（英文）,2026,6(1):73-90,18.基金项目
This work is supported by the National Natural Science Foundation of China(82174076),the Construction Project of Liaoning Provincial Key Laboratory,China(2022JH13/10200026),the Natural Science Foundation of Hebei Province(H2024501002),the Fundamental Research Funds for the Central Universities(N2423006),and the 111 Project(B16009). （82174076）
