中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (17): 2681-2686.doi: 10.12307/2023.423

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    

铁死亡参与高蛋氨酸饮食诱导ApoE-/-小鼠肝损伤的发病过程

李媛媛1，孙  岳1，宝  瑞1，畅思容1，王  梦1，余梦雪1，杨安宁2，刘志宏1   

  1. 宁夏医科大学，1公共卫生与管理学院，2基础医学院，宁夏回族自治区银川市  750004
  • 收稿日期:2022-06-06 接受日期:2022-07-21 出版日期:2023-06-18 发布日期:2022-10-24
  • 通讯作者: 刘志宏，博士，教授，宁夏医科大学，公共卫生与管理学院，宁夏回族自治区银川市 750004 杨安宁，博士，副教授，宁夏医科大学，基础医学院，宁夏回族自治区银川市 750004
  • 作者简介:李媛媛，女，1995年生，河南省淇县人，汉族，宁夏医科大学在读硕士，主要从事环境、职业危害与健康研究。
  • 基金资助:
    国家自然科学基金(81960018)，项目负责人：刘志宏；国家自然科学基金(82060264)，项目负责人：孙岳；国家自然科学基金(82160088)，项目负责人：杨安宁；宁夏回族自治区科学技术厅重点研发计划项目(2020BEG03008)，项目负责人：孙岳；宁夏回族自治区科学技术厅重点研发计划项目(2020BFH02003)，项目负责人：杨安宁；宁夏回族自治区科学技术厅重点研发计划项目(2021BEG02030)，项目负责人：刘志宏

Ferroptosis is involved in the pathogenesis of liver injury induced by high methionine diet in ApoE-/- mice

Li Yuanyuan1, Sun Yue1, Bao Rui1, Chang Sirong1, Wang Meng1, Yu Mengxue1, Yang Anning2, Liu Zhihong1   

  1. 1School of Public Health and Management, 2School of Basic Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Received:2022-06-06 Accepted:2022-07-21 Online:2023-06-18 Published:2022-10-24
  • Contact: Liu Zhihong, MD, Professor, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China Yang Anning, MD, Associate professor, School of Basic Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Li Yuanyuan, Master candidate, School of Public Health and Management, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    the National Natural Science Foundation of China, Nos. 81960018 (to LZH), 82060264 (to SY), and 82160088 (to YAN); Key R&D Project of the Department of Science and Technology of Ningxia Hui Autonomous Region, Nos. 2020BEG03008 (to SY), 2020BFH02003 (to YAN), and 2021BEG02030 (to LZH) 

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摘要：

文题释义：

铁死亡：是一种依赖于细胞内铁的调节性细胞死亡模式，并伴随着高水平的脂质过氧化，形态学特征为线粒体体积缩小、线粒体膜密度增高和线粒体减少或消失。铁死亡的主要发生机制是由于铁离子的蓄积引起细胞中活性氧异常增多，导致细胞膜通透性丧失，最终引起细胞死亡。
高同型半胱氨酸血症：是一种以血液中同型半胱氨酸升高为特征的疾病，通常指同型半胱氨酸> 15 µmol/L，被认为是引起肝脏疾病、心血管疾病以及癌症等多种疾病的独立危险因素。

背景：高同型半胱氨酸可以通过氧化应激、炎症反应和内质网应激等机制促进肝损伤的发生。高蛋氨酸饮食诱导的E型载脂蛋白基因敲除(ApoE-/-)小鼠可以引起肝损伤以及体内同型半胱氨酸水平升高，铁死亡是一种依赖于细胞内的铁并引起脂质过氧化物过量蓄积的细胞调节性死亡途径。然而，它是否参与了高蛋氨酸饮食诱导肝损害的形成需进一步研究和探讨。
目的：探讨铁死亡在ApoE-/-小鼠高同型半胱氨酸致肝损伤中的作用。
方法：12只6-8周龄雄性ApoE-/-小鼠，体质量20-25 g，随机分为对照组和高蛋氨酸组，每组6只，分别采用普通饲料和高蛋氨酸饲料喂养13.5周。喂养结束后通过病理学观察以及肝组织中天冬氨酸转氨酶、丙氨酸转氨酶活性检测评估小鼠肝损伤情况；组织铁检测试剂盒测定小鼠肝组织中的铁离子含量；通过肝组织中丙二醛含量和荧光强度评估小鼠肝组织中脂质过氧化程度；实时荧光定量PCR与Western blotting 方法分别检测两组小鼠肝组织中P53和谷胱甘肽过氧化物酶4的mRNA和蛋白表达水平。
结果与结论：①与对照组相比，高蛋氨酸组发现大量肝细胞排列紊乱，间隙增大，胞浆疏松的典型肝损伤组织病理学改变；天冬氨酸转氨酶以及丙氨酸转氨酶活性升高(P < 0.01)；肝组织中铁离子含量增加(P < 0.01)，丙二醛含量(P < 0.01)及荧光强度均增加；②实时荧光定量PCR与Western blotting检测发现，高蛋氨酸组小鼠肝组织中谷胱甘肽过氧化物酶4的表达水平降低(P < 0.01)，P53的表达水平升高(P < 0.05)；③提示铁死亡参与高蛋氨酸饮食诱导ApoE-/-小鼠肝损伤的发病过程。

https://orcid.org/0000-0002-6270-7998(李媛媛)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 肝损伤, 铁死亡, 脂质过氧化, 高同型半胱氨酸血症, 丙二醛

Abstract: BACKGROUND: Homocysteine can promote the occurrence of liver injury through oxidative stress, inflammatory response, and endoplasmic reticulum stress. High methionine diet-induced apolipoprotein gene knockout (ApoE-/-) can induce liver injury and increase in vivo homocysteine level in mice. Ferroptosis is a cellular regulatory death pathway that depends on intracellular iron and causes excessive accumulation of lipid peroxides. However, whether it is involved in the formation of liver injury induced by high methionine diet needs to be further studied and discussed.
OBJECTIVE: To explore the role of ferroptosis in hyperhomocysteine-induced liver injury in ApoE-/- mice. 
METHODS: Twelve ApoE-/- mice aged 6 to 8 weeks were randomly divided into control group and high methionine group (n=6 per group), and fed with normal diet and high methionine diet for 13.5 weeks. Liver injury in ApoE-/- mice was evaluated by pathological observation and detection of aspartate aminotransferase and alanine aminotransferase activity in liver tissue. Iron ion concentration in liver tissue of mice was tested by tissue iron detection kit. Malondialdehyde content and fluorescence intensity were detected to evaluate the degree of lipid peroxidation in the liver tissue of mice. Real-time fluorescence quantitative PCR and western blot were performed to determine the expression of P53 and glutathione peroxidase 4 at mRNA and protein levels in the liver tissue. 
RESULTS AND CONCLUSION: Compared with the control group, a large number of liver cells in the high methionine diet group were found to have the typical histopathological changes of liver injury, such as disordered arrangement of liver cells, enlarged space, and loose cytoplasm. Compared with the control group, the activities of aspartate aminotransferase and alanine aminotransferase were significantly increased (P < 0.01). Meanwhile, the concentration of iron ions (P < 0.01), malondialdehyde content (P < 0.01), and fluorescence intensity in liver tissue were also significantly increased in the high methionine diet group. Real-time fluorescence quantitative PCR and western blot results showed that the expression level of glutathione peroxidase 4 was decreased (P < 0.01) and the expression of P53 was increased (P < 0.05) in ApoE-/- mice in the high methionine diet group. These findings indicate that ferroptosis is involved in the pathogenesis of liver injury in ApoE-/- mice induced by high methionine diet.

Key words: liver injury, ferroptosis, lipid peroxidation, hyperhomocysteinemia, malondialdehyde

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