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高原低氧暴露后小鼠海马铁死亡相关通路的变化

常熙雯^1,2, 赵安鹏^1,2, 钟艳², 刘菲菲², 王荣^2,*

¹兰州大学药学院，兰州 730000；²中国人民解放军联勤保障部队第九四〇医院药剂科，兰州 730050

摘要

本文旨在研究高原低氧暴露后小鼠海马组织铁死亡的发生情况及相关通路的变化。采用海拔4 010 m的高原环境建立低压性缺氧模型，用HE染色观察小鼠海马组织形态学变化，用免疫组织化学染色法观察海马组织脂质过氧化水平，用试剂盒测定海马组织丙二醛(malondialdehyde, MDA)、还原型谷胱甘肽(glutathione, GSH)和Fe2+水平，用Western blot检测海马组织中谷胱甘肽过氧化物酶4 (glutathione peroxidase 4, GPX4)、溶质载体家族7成员11 (solute carrier family 7 member 11, SLC7A11)、铁蛋白重链1 (ferritin heavy chain 1, FTH1)、转铁蛋白1 (ferroportin 1, FPN1)、转铁蛋白受体1 (transferrin receptor 1, TfR1)、铁死亡抑制蛋白1 (ferroptosis suppressor protein 1, FSP1)和长链脂酰辅酶A合成酶4 (acyl-CoA synthetase long chain family member 4, ACSL4)蛋白表达水平。结果显示，与平原对照组比较，高原低氧暴露1、3、7和14 d组小鼠海马齿状回均出现明显的病理损伤，排列紊乱，核固缩明显。与平原对照组比较，高原低氧暴露增加小鼠海马齿状回4-羟基壬烯醛(4- hydroxynonenal, 4-HNE)和海马MDA含量，显著降低海马GSH含量；高原低氧暴露14 d组小鼠海马Fe2+含量显著增加。与平原对照组比较，高原低氧暴露小鼠海马GPX4、FTH1、FPN1、TfR1和FSP1蛋白表达水平均显著下调，SLC7A11蛋白表达水平仅在高原低氧暴露7 d组显著下调，ACSL4蛋白表达水平仅在高原低氧暴露14 d组显著上调。以上结果提示，高原低氧暴露14 d可减少小鼠海马GSH合成，下调GPX4表达，导致GSH代谢紊乱，抑制铁储存和铁外排，促进脂质过氧化反应发生，同时抑制还原型辅酶Q10 (CoQ10H2)抗脂质过氧化作用，这可能导致小鼠海马组织中细胞铁死亡发生。

关键词： 高原低氧; 铁死亡; 谷胱甘肽代谢; 铁代谢; 脂质过氧化

Changes of ferroptosis related pathways in hippocampus of mice exposed to high-altitude hypoxia

CHANG Xi-Wen^1,2, ZHAO An-Peng^1,2, ZHONG Yan², LIU Fei-Fei², WANG Rong^2,*

¹School of Pharmacy, Lanzhou University, Lanzhou 730000, China；²Department of Pharmacy, The 940th Hospital of PLA Joint Logistics Support Force, Lanzhou 730050, China

Abstract

The present study aimed to investigate the occurrence of ferroptosis in mouse hippocampal tissue and changes in related pathways after exposure to high-altitude hypoxia. A low-pressure hypoxia model was established using a high-altitude environment at 4 010 m. HE staining was used to observe morphological changes in mouse hippocampal tissue, immunohistochemical staining was used to observe lipid peroxidation levels in hippocampal tissue, and corresponding kits were used to measure malondialdehyde (MDA), reduced glutathione (GSH), and Fe2+ levels in hippocampal tissue. Western blot was used to detect glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), ferritin heavy chain 1 (FTH1), ferroportin 1 (FPN1), transferrin receptor 1 (TfR1), ferroptosis suppressor protein 1 (FSP1), and acyl-CoA synthase long chain family member 4 (ACSL4). The results showed that, compared with the plain control group, the mice exposed to high-altitude hypoxia for 1, 3, 7, and 14 d exhibited significant pathological damage, disordered arrangement, and obvious nuclear condensation in the dentate gyrus of the hippocampus. Compared with the plain control group, high-altitude hypoxia exposure increased 4-hydroxynonenal (4-HNE) content in the dentate gyrus and hippocampal MDA content, whereas significantly decreased hippocampal GSH content. Compared with the plain control group, the Fe2+ content in the hippocampus of mice exposed to high-altitude hypoxia for 14 d significantly increased. Compared with the plain control group, the protein expression levels of GPX4, FTH1, FPN1, TfR1, and FSP1 in the hippocampus of mice exposed to high-altitude hypoxia were significantly down-regulated (SLC7A11 was significantly down-regulated only in the 7-d high-altitude hypoxia exposure group), while the protein expression level of ACSL4 was only significantly up-regulated in the 14-d high-altitude hypoxia exposure group. These results suggest that exposure to high-altitude hypoxia for 14 d can reduce GSH synthesis in mouse hippocampus, down-regulate GPX4 expression, lead to GSH metabolism disorders, inhibit iron storage and efflux, promote lipid peroxidation reaction, and inhibit CoQ10H2’s anti-lipid peroxidation effect, ultimately leading to ferroptosis.

Key words: high-altitude hypoxia; ferroptosis; glutathione metabolism; iron metabolism; lipid peroxidation

收稿日期：　　录用日期：

通讯作者：王荣　　E-mail:

DOI: 10.13294/j.aps.2024.0057

引用本文：

常熙雯, 赵安鹏, 钟艳, 刘菲菲, 王荣. 高原低氧暴露后小鼠海马铁死亡相关通路的变化[J]. 生理学报 2024; 76 (4): 507-516.

CHANG Xi-Wen, ZHAO An-Peng, ZHONG Yan, LIU Fei-Fei, WANG Rong. Changes of ferroptosis related pathways in hippocampus of mice exposed to high-altitude hypoxia. Acta Physiol Sin 2024; 76 (4): 507-516 (in Chinese with English abstract).