Department of Neurosurgery, the 1st Affiliated Hospital of Chongqing Medical University, Chong Qing, China.
Department of Experimental and Clinical Neuropathology, Mossakowski Medical Research Centre Polish Academy of Sciences, Warsaw, Poland.
Oxid Med Cell Longev. 2021 Dec 27;2021:9800794. doi: 10.1155/2021/9800794. eCollection 2021.
Iron metabolism disturbances play an important role in early brain injury (EBI) after subarachnoid hemorrhage (SAH), and hepcidin largely influences iron metabolism. Importantly, iron metabolism may be associated with ferroptosis, recently a nonapoptotic iron-dependent form of cell death that may have a great impact on brain injury after SAH. We investigated hepcidin on iron metabolism and ferroptosis involving divalent metal transporter 1 (DMT1), and ferroportin-1 (FPN1) in a rat model of SAH. Male Sprague-Dawley rats were subjected to the endovascular perforation to induce SAH, and treated with heparin (inhibitor of hepcidin), or oncostatin M (OSM, inducer of hepcidin), or ebselen (inhibitor of DMT1) by intracerebroventricular injections. Hepcidin, DMT1, FPN1 and glutathione peroxidase 4 (GPX4), were detected by western blot and immunofluorescence. Iron metabolism was detected through Perl's iron staining and iron content assay. Ferroptosis, the ROS production, lipid peroxidation (LPO) was evaluated by monitoring methane dicarboxylic aldehyde (MDA), glutathione (GSH), glutathione peroxidase 4 (GPX4) activity, and transmission electron microscopy. Neurological deficit scores, Evans blue staining and brain water content were also determined to detect EBI 72 h after SAH. Our results showed that inhibition of DMT1 by ebselen could suppress iron accumulation and lipid peroxidation, and thereby alleviate ferroptosis and EBI in SAH rats. Heparin downregulated the expression of hepcidin and DMT1, increased FPN1, and exerted protective effects that were equivalent to those of ebselen on ferroptosis and EBI. In addition, OSM increased the expression of hepcidin and DMT1, decreased FPN1, and aggravated ferroptosis and EBI, while the effect on ferroptosis was reversed by ebselen. Therefore, the study revealed that hepcidin could regulate iron metabolism and contribute to ferroptosis via DMT1 signaling activation in rats with EBI after SAH.
铁代谢紊乱在蛛网膜下腔出血 (SAH) 后早期脑损伤 (EBI) 中起重要作用，hepcidin 对铁代谢有很大影响。重要的是，铁代谢可能与铁死亡有关，铁死亡是一种最近发现的非凋亡性、铁依赖性细胞死亡形式，可能对 SAH 后脑损伤有重大影响。我们在 SAH 大鼠模型中研究了 hepcidin 对涉及二价金属转运蛋白 1 (DMT1) 和铁蛋白-1 (FPN1) 的铁代谢和铁死亡的影响。雄性 Sprague-Dawley 大鼠接受血管内穿孔诱导 SAH，并通过脑室内注射肝素（hepcidin 抑制剂）、抑瘤素 M（hepcidin 诱导剂）或依布硒啉（DMT1 抑制剂）进行治疗。通过 Western blot 和免疫荧光检测 hepcidin、DMT1、FPN1 和谷胱甘肽过氧化物酶 4 (GPX4)。通过 Perl 铁染色和铁含量测定检测铁代谢。通过监测甲烷二羧酸醛 (MDA)、谷胱甘肽 (GSH)、谷胱甘肽过氧化物酶 4 (GPX4) 活性和透射电子显微镜评估铁死亡、ROS 产生和脂质过氧化 (LPO)。还测定了神经功能缺损评分、伊文思蓝染色和脑水含量，以检测 SAH 后 72 小时的 EBI。结果表明，依布硒啉抑制 DMT1 可抑制铁积累和脂质过氧化，从而减轻 SAH 大鼠的铁死亡和 EBI。肝素下调 hepcidin 和 DMT1 的表达，增加 FPN1，并发挥与依布硒啉相当的保护作用，减轻铁死亡和 EBI。此外，OSM 增加了 hepcidin 和 DMT1 的表达，降低了 FPN1，并加重了铁死亡和 EBI，而依布硒啉则逆转了铁死亡的作用。因此，该研究表明，hepcidin 通过激活 DMT1 信号通路调节铁代谢，导致 SAH 后 EBI 大鼠发生铁死亡。
