He Yifan, Yi Tingting, Yao Ying, Duan Songyuan, Wang Lianhang, Gao Jianmei, Gong Qihai
Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi, China; Department of Pharmacology, Key Laboratory of Basic Pharmacology of Guizhou Province and School of Pharmacy, Zunyi Medical University, Zunyi, Guizhou, China.
Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi, China; Department of Pharmacology, Key Laboratory of Basic Pharmacology of Guizhou Province and School of Pharmacy, Zunyi Medical University, Zunyi, Guizhou, China.
Free Radic Biol Med. 2025 Dec 16;241:353-366. doi: 10.1016/j.freeradbiomed.2025.09.044. Epub 2025 Sep 23.
Copper overload triggers cuproptosis, a copper-dependent cell death pathway characterized by mitochondrial oxidative stress, dysfunction, and disrupted dynamics, posing significant threats to neuronal health. Icaritin (ICT), a bioactive flavonoid from Herbal Epimedii, exhibits antioxidant and neuroprotective properties, but its impact on cuproptosis remains unexplored. Thus, this study was aimed to investigate ICT's protective mechanisms against cuproptosis induced by the cupric sulfate and copper ionophore elesclomol (Cu-ES) in HT22 hippocampal neuronal cells. We found that Cu-ES effectively modeled cuproptosis, reducing viability by 50 % and inducing severe mitochondrial damage, oxidative stress, dysfunction, tricarboxylic acid cycle disruption, and dynamics imbalance. While ICT's treatment concentration-dependently mitigated these injuries. Mechanistically, computational molecular interaction analysis and trajectory simulations, and surface plasmon resonance confirmed ICT directly binds ferredoxin 1 (FDX1) with high affinity and stability, downregulating its protein expression. ICT consequently inhibited the FDX1-mediated cuproptosis pathway, reducing dihydrolipoamide S-acetyltransferase (DLAT) oligomerization, modulating cuproptosis sensitivity proteins, restoring copper homeostasis by increasing ATPase copper transporting beta (ATP7B) and decreasing solute carrier family 31 member 1 (SLC31A1), and suppressing the lipoylation pathway. Crucially, FDX1 knockdown abolished Cu-ES toxicity and potentiated ICT's protective effects against superoxide production, DLAT expression, and copper accumulation. Furthermore, ICT rescued mitochondrial dynamics by promoting fusion and inhibiting fission. Our findings demonstrate ICT is a potent inhibitor of neuronal cuproptosis, targeting FDX1 to alleviate mitochondrial oxidative stress, dysfunction, and dynamics disorder, presenting a promising therapeutic strategy.
铜过载引发铜死亡，这是一种依赖铜的细胞死亡途径，其特征为线粒体氧化应激、功能障碍和动力学紊乱，对神经元健康构成重大威胁。淫羊藿苷（ICT）是一种来源于淫羊藿的生物活性黄酮类化合物，具有抗氧化和神经保护特性，但其对铜死亡的影响尚未得到探索。因此，本研究旨在探讨ICT对硫酸酮和铜离子载体依斯氯铵（Cu-ES）诱导的HT22海马神经元细胞铜死亡的保护机制。我们发现，Cu-ES有效地模拟了铜死亡，使细胞活力降低50%，并导致严重的线粒体损伤、氧化应激、功能障碍、三羧酸循环中断和动力学失衡。而ICT的治疗可浓度依赖性地减轻这些损伤。从机制上讲，计算分子相互作用分析、轨迹模拟和表面等离子体共振证实，ICT以高亲和力和稳定性直接结合铁氧还蛋白1（FDX1），下调其蛋白表达。因此，ICT抑制了FDX1介导的铜死亡途径，减少二氢硫辛酰胺S-乙酰转移酶（DLAT）寡聚化，调节铜死亡敏感性蛋白，通过增加ATP酶铜转运β（ATP7B）和减少溶质载体家族31成员1（SLC31A1）来恢复铜稳态，并抑制脂酰化途径。至关重要的是，敲低FDX1消除了Cu-ES的毒性，并增强了ICT对超氧化物产生、DLAT表达和铜积累的保护作用。此外，ICT通过促进融合和抑制裂变来挽救线粒体动力学。我们的研究结果表明，ICT是一种有效的神经元铜死亡抑制剂，靶向FDX1以减轻线粒体氧化应激、功能障碍和动力学紊乱，是一种有前景的治疗策略。
