Chen Xi, Liu Li, Jiang Wu, Hu Yu, Zou Wei, Zhang Ping, Wang Bo
Department of Neurology, The Affiliated Nanhua Hospital, Hengyang Medical School, University of South China, Hengyang, Hunan, China.
Institute of Clinical Laboratory, The First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, Hunan, China.
J Physiol Investig. 2025 Mar 1;68(2):91-99. doi: 10.4103/ejpi.EJPI-D-24-00099. Epub 2025 Mar 19.
Our previous research has established that hydrogen sulfide (H 2 S) exerts an antagonistic effect against the hippocampal neurotoxicity induced by Rotenone (ROT). However, the underlying mechanisms are so far poorly understood. Substantial evidence corroborates the involvement of ferroptosis in ROT-induced neurotoxicity. To elucidate the protective mechanism of H 2 S against ROT-induced hippocampal neurotoxicity, this study explores its regulatory role in ferroptosis and its underlying mechanisms. We used Fluoro-Jade B staining to detect dead neurons. The levels of ferrous ions and glutathione (GSH) were measured by a kit. The ferroptosis-related proteins, including light-chain subunit (xCT), GSH peroxidase 4(GPX4), ferroptosis marker acyl-CoA synthetase long-chain family member 4(ACSL4), and ferritinophagy-related protein, including ferritin heavy chain 1 (FTH1), sequestosome 1 (p62), ferritinophagy markers autophagosome marker light-chain I/II (LC3I/II), and nuclear receptor coactivator 4 (NCOA4), were measured by Western blot. Our findings indicate that H 2 S reduces hippocampal neuron deaths in ROT-exposed rats. Meanwhile, H 2 S reverses the downregulations of xCT and GPX4, and the upregulations of ferrous ion and ACSL4 in the hippocampus induced by ROT. Furthermore, H 2 S reverses the upregulations of LC3I/II and NCOA4, and the downregulations of P62 and FTH1. Based on these findings, we concluded that the protective role of H 2 S against ROT-induced hippocampal neuronal death involves inhibiting ferroptosis triggered by ferritinophagy.
我们之前的研究已经证实，硫化氢（H₂S）对鱼藤酮（ROT）诱导的海马神经毒性具有拮抗作用。然而，其潜在机制目前仍知之甚少。大量证据证实铁死亡参与了ROT诱导的神经毒性。为了阐明H₂S对ROT诱导的海马神经毒性的保护机制，本研究探讨了其在铁死亡中的调节作用及其潜在机制。我们使用Fluoro-Jade B染色来检测死亡神经元。通过试剂盒测量亚铁离子和谷胱甘肽（GSH）的水平。通过蛋白质免疫印迹法检测铁死亡相关蛋白，包括轻链亚基（xCT）、谷胱甘肽过氧化物酶4（GPX4）、铁死亡标志物酰基辅酶A合成酶长链家族成员4（ACSL4），以及铁自噬相关蛋白，包括铁蛋白重链1（FTH1）、聚集体蛋白1（p62）、铁自噬标志物自噬体标志物轻链I/II（LC3I/II）和核受体辅激活因子4（NCOA4）。我们的研究结果表明，H₂S可减少ROT处理大鼠海马神经元的死亡。同时，H₂S可逆转ROT诱导的海马中xCT和GPX4的下调，以及亚铁离子和ACSL4的上调。此外，H₂S可逆转LC3I/II和NCOA4的上调，以及P62和FTH1的下调。基于这些发现，我们得出结论，H₂S对ROT诱导的海马神经元死亡的保护作用涉及抑制铁自噬引发的铁死亡。
