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膜联蛋白A1模拟肽Ac2-26对RSL3诱导的人脐静脉内皮细胞铁死亡及线粒体功能的影响
作者:
作者单位:

(1. 南华大学心血管疾病研究所 动脉硬化学湖南省重点实验室 湖南省动脉硬化性疾病国际科技创新合作基地,湖南省衡阳市 421001;2.南华大学附属第一医院,湖南省衡阳市 421001)

作者简介:

谈世铭,硕士研究生,研究方向为动脉粥样硬化发病学与防治研究,E-mail:719582339@qq.com。通信作者王佐,博士,教授,博士研究生导师,研究方向为动脉粥样硬化发病学与防治研究,E-mail:smt121101@163.com。

基金项目:

湖南省自然科学基金项目(2024JJ9395)


Effects of annexin A1 mimic peptide Ac2-26 on ferroptosis and mitochondrial function of human umbilical vein endothelial cells induced by RSL3
Author:
  • TAN Shiming 1

    TAN Shiming

    Institute of Cardiovascular Disease, University of South China & Key Laboratory of Arterial Hard Chemistry of Hunan Province & Hunan Province Arteriosclerotic Disease International Scientific and Technological Innovation Cooperation Base, Hengyang, Hunan 421001, China
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  • CAO Zitong 1

    CAO Zitong

    Institute of Cardiovascular Disease, University of South China & Key Laboratory of Arterial Hard Chemistry of Hunan Province & Hunan Province Arteriosclerotic Disease International Scientific and Technological Innovation Cooperation Base, Hengyang, Hunan 421001, China
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  • WANG Jingjing 1

    WANG Jingjing

    Institute of Cardiovascular Disease, University of South China & Key Laboratory of Arterial Hard Chemistry of Hunan Province & Hunan Province Arteriosclerotic Disease International Scientific and Technological Innovation Cooperation Base, Hengyang, Hunan 421001, China
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  • JIA Jinqiu 1

    JIA Jinqiu

    Institute of Cardiovascular Disease, University of South China & Key Laboratory of Arterial Hard Chemistry of Hunan Province & Hunan Province Arteriosclerotic Disease International Scientific and Technological Innovation Cooperation Base, Hengyang, Hunan 421001, China
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  • LI Keyi 1

    LI Keyi

    Institute of Cardiovascular Disease, University of South China & Key Laboratory of Arterial Hard Chemistry of Hunan Province & Hunan Province Arteriosclerotic Disease International Scientific and Technological Innovation Cooperation Base, Hengyang, Hunan 421001, China
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  • CAI Zemin 2

    CAI Zemin

    The First Affiliated Hospital of University of South China, Hengyang, Hunan 421001, China
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  • WANG Zuo 1

    WANG Zuo

    Institute of Cardiovascular Disease, University of South China & Key Laboratory of Arterial Hard Chemistry of Hunan Province & Hunan Province Arteriosclerotic Disease International Scientific and Technological Innovation Cooperation Base, Hengyang, Hunan 421001, China
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Affiliation:

1.Institute of Cardiovascular Disease, University of South China & Key Laboratory of Arterial Hard Chemistry of Hunan Province & Hunan Province Arteriosclerotic Disease International Scientific and Technological Innovation Cooperation Base, Hengyang, Hunan 421001, China;2.The First Affiliated Hospital of University of South China, Hengyang, Hunan 421001, China)

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    摘要:

    目的]探究膜联蛋白A1模拟肽Ac2-26对人脐静脉内皮细胞(HUVEC)铁死亡的影响及其机制。 [方法]用经典铁死亡激动剂RSL3诱发HUVEC铁死亡,然后用膜联蛋白A1模拟肽Ac2-26进行干预。CCK-8试剂盒检测细胞数量与活力,ELISA检测丙二醛(MDA)和谷胱甘肽(GSH)水平,Western blot检测铁死亡相关分子及黏附分子表达,C11-BODIPY荧光探针检测脂质活性氧(ROS)水平,MitoSOX检测线粒体活性氧(mtROS)水平,FeRhoNOX-1荧光探针检测细胞内Fe2＋含量,透视显微镜观察线粒体形态,JC-1荧光探针检测线粒体膜电位,试剂盒检测ATP含量,划痕实验检测细胞迁移能力,硝酸还原酶法检测一氧化氮(NO)水平。 [结果]Ac2-26抑制RSL3诱导的HUVEC活力下降,上调抑制铁死亡蛋白溶质载体家族7成员11(SLC7A11)、GPX4和铁蛋白重链1(FTH1)的表达,增加GSH含量,降低MDA含量,减少细胞内脂质ROS生成,降低细胞内Fe2＋聚集(P＜0.05或P＜0.01)；Ac2-26抑制RSL3诱导HUVEC线粒体形态和功能的损伤,上调ATP含量(P＜0.05)及线粒体膜电位(P＜0.001)；Ac2-26抑制RSL3诱导的HUVEC迁移能力的下降,上调NO水平,抑制细胞间黏附分子1(ICAM-1)及白细胞介素1β(IL-1β)的蛋白表达(P＜0.05或P＜0.01)。 [结论]Ac2-26抑制RSL3诱导的HUVEC铁死亡,并维护线粒体形态与功能及HUVEC功能。

    Abstract:

    Aim To explore the effect and mechanism of annexin A1 mimic peptide Ac2-26 on ferroptosis in human umbilical vein endothelial cells (HUVEC). Methods Induction of HUVEC ferroptosis was achieved by the classical ferroptosis agonist RSL3, with subsequent intervention by the annexin A1 mimtic peptide Ac2-26. The cell number and viability were detected by CCK-8 kit, the levels of malondialdehyde (MDA) and glutathione (GSH) were detected by ELISA, the expression of ferroptosis-related molecules and adhesion molecules was detected by Western blot, the lipid reactive oxygen species (ROS) levels were detected by C11-BODIPY fluorescent probe, and the mitochondrial reactive oxygen species (mtROS) levels were detected by MitoSOX probe. FeRhoNOX-1 fluorescent probe was used to detect intracellular Fe2＋ content, perspective microscopy was used to observe mitochondrial morphology, JC-1 fluorescent probe was used to detect mitochondrial membrane potential, kit was used to detect ATP levels, the Scratch assay was used to detect cell migration ability, and nitrate reductase assay was used to detect nitric oxide (NO) level. Results Ac2-26 inhibited RSL3-induced decrease in HUVEC viability, up-regulated the expression of suppressor of ferroptosis proteolytic carrier family 7 member 11 (SLC7A11), GPX4, and ferritin heavy chain 1 (FTH1), increased the GSH content, decreased the MDA content, reduced the generation of intracellular lipid ROS, and decreased the intracellular Fe2＋ aggregation (P＜0.05 or P＜0.01); Ac2-26 inhibited RSL3-induced damage to HUVEC mitochondrial morphology and function, up-regulated ATP content (P＜0.05) and mitochondrial membrane potential (P＜0.001); Ac2-26 inhibited RSL3-induced decrease in HUVEC migratory ability, up-regulated NO levels, inhibited intercellular adhesion molecule-1 (ICAM-1) and interleukin-1β (IL-1β) protein expression (P＜0.05 or P＜0.01). Conclusion Ac2-26 inhibits RSL3-induced ferroptosis in HUVEC and maintains mitochondrial morphology and function, as well as HUVEC function.

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谈世铭,曹子彤,王晶晶,贾金秋,李珂怡,蔡泽民,王佐.膜联蛋白A1模拟肽Ac2-26对RSL3诱导的人脐静脉内皮细胞铁死亡及线粒体功能的影响[J].中国动脉硬化杂志,2025,33(4):303~309, 341.

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  • 收稿日期:2024-10-25
  • 最后修改日期:2025-03-18
  • 在线发布日期: 2025-05-16

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