Article(id=1276976084767867325, tenantId=1146029695717560320, journalId=1276577828834349082, issueId=1276975955985961244, articleNumber=null, orderNo=null, doi=10.19540/j.cnki.cjcmm.20250902.401, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1747584000000, receivedDateStr=2025-05-19, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1782384423676, onlineDateStr=2026-06-25, pubDate=1767196800000, pubDateStr=2026-01-01, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782384423676, onlineIssueDateStr=2026-06-25, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782384423676, creator=13701087609, updateTime=1782384423676, updator=13701087609, issue=Issue{id=1276975955985961244, tenantId=1146029695717560320, journalId=1276577828834349082, year='2026', volume='51', issue='1', pageStart='1', pageEnd='300', issueExtLink='null', onlineDate='null', pubDate='1767196800000', pubDateStr='2026-01-01', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782384392973, creator='13701087609', updateTime=1782384484405, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1276976339546673871, tenantId=1146029695717560320, journalId=1276577828834349082, issueId=1276975955985961244, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1276976339546673872, tenantId=1146029695717560320, journalId=1276577828834349082, issueId=1276975955985961244, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=191, endPage=201, ext={EN=ArticleExt(id=1276976085107605951, articleId=1276976084767867325, tenantId=1146029695717560320, journalId=1276577828834349082, language=EN, title=Salidroside suppresses ferroptosis in HT22 cells following oxygen-glucose deprivation/reoxygenation through regulation of non-ubiquitinated FUNDC1-dependent mitophagy pathway, columnId=null, journalTitle=Chinese Journal of Traditional Chinese Medicine, columnName=null, runingTitle=null, highlight=null, articleAbstract=

This study aims to investigate whether the mitophagy receptor FUN14 domain-containing 1 (FUNDC1) serves as a molecular link between mitophagy and neuronal ferroptosis, and to determine whether salidroside (Sal) can inhibit neuronal ferroptosis after oxygen-glucose deprivation/reoxygenation (OGD/R) by inducing FUNDC1 expression and regulating mitophagy, thereby exerting neuroprotective effects. An in vitro model of neuronal ischemia-reperfusion injury was established with HT22 cells subjected to OGD/R. The experiment consisted of three parts: ① control, OGD/R, ferrostatin-1 (Fer-1), FUNDC1 overexpression (OV-FUNDC1), and OV-FUNDC1+Fer-1 groups; ② control, OGD/R, 3-methyladenine (3MA, an autophagy inhibitor), Sal, and Sal+3MA groups; ③control, OGD/R, Sal, FUNDC1 silencing (Si-FUNDC1), and Sal+Si-FUNDC1 groups. In the first part of the experiment, the survival rate of cells in each group was detected by the CCK-8 assay, and the protein levels of p62, microtubule-associated protein light chain 3 (LC3), acyl-CoA synthetase long-chain family 4 (ACSL4), and glutathione peroxidase 4 (GPX4) were measured by Western blot. The results showed that compared with the control group, the OGD/R group had down-regulated protein levels of p62 and GPX4 (P<0.05, P<0.01) and up-regulated protein level of ACSL4 (P<0.01); compared with the OGD/R group, the OV-FUNDC1 group showed increased protein levels of LC3 and GPX4 (P<0.05, P<0.01) and decreased protein levels of p62 and ACSL4 (P<0.05, P<0.01). In the second part of the experiment, after Sal intervention, the protein levels of FUNDC1, p62, LC3, ACSL4, and GPX4 were determined by Western blot, and changes in mitochondrial membrane potential were measured via JC-1. The results showed that compared with the OGD/R group, the Sal group had up-regulated protein levels of FUNDC1, LC3, and GPX4 (P<0.01), down-regulated protein levels of p62 and ACSL4 (P<0.01), and increased mitochondrial membrane potential (P<0.01); the 3MA group showed decreased mitochondrial membrane potential (P<0.01). In the third part of the experiment, after silencing of FUNDC1, the intracellular Fe²⁺ content was measured via a ferrous ion assay kit; reactive oxygen species (ROS) levels were measured by flow cytometry; the mitochondrial function was assessed via MitoTracker Red; adenosine triphosphate (ATP) and glutathione (GSH) levels were detected using assay kits; the expression of ACSL4 and GPX4 was detected by immunofluorescence; the protein levels of p62, LC3, ACSL4, and GPX4 were measured by Western blot. The results showed that compared with the Si-FUNDC1 group, the Sal group had decreased Fe²⁺ concentration (P<0.01), reduced ROS level (P<0.01), increased MitoTracker Red fluorescence intensity (P<0.01) and GSH content (P<0.01), weakened ACSL4 fluorescence intensity (P<0.01), enhanced GPX4 fluorescence intensity (P<0.01), up-regulated protein levels of LC3 and GPX4 (P<0.01), and down-regulated protein levels of p62 and ACSL4 (P<0.01). This study reveals that FUNDC1 may be a key protein linking mitochondrial autophagy and ferroptosis in neurons after ischemia-reperfusion injury, and Sal intervention can inhibit neuronal ferroptosis after ischemia-reperfusion by promoting the non-ubiquitinated FUNDC1-dependent mitophagy pathway.

, authors=null, authorsList=Qiu-si DU, Jun LIAO, authorCompany=null, correspAuthors=Jun LIAO, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1276976085480899009, articleId=1276976084767867325, tenantId=1146029695717560320, journalId=1276577828834349082, language=CN, title=红景天苷调控非泛素化FUNDC1线粒体自噬途径抑制HT22细胞糖氧剥夺／复氧后铁死亡, columnId=1276975994707775974, journalTitle=中国中药杂志, columnName=药理, runingTitle=null, highlight=null, articleAbstract=

基于线粒体自噬受体FUN14结构域蛋白1（FUNDC1）是否是线粒体自噬与神经元铁死亡的关联靶点，探讨红景天苷（Sal）是否可以通过诱导FUNDC1的表达，调控线粒体自噬从而抑制氧糖剥夺／复氧（OGD/R）后神经元铁死亡，起到神经系统保护作用。建立HT22细胞ODG/R培养制作神经元缺血再灌注损伤模型，进行3部分实验：①对照组、OGD/R组、铁抑素（Fer-1）组、过表达FUNDC1（OV-FUNDC1）组、OV-FUNDC1+Fer-1组；②对照组、OGD/R组、自噬抑制剂3-甲基腺嘌呤（3MA）组、Sal组、Sal+3MA组；③对照组、OGD/R组、Sal组、沉默FUNDC1（Si-FUNDC1）组、Sal+Si-FUNDC1组。第一部分实验，CCK-8法检测各组细胞存活率；蛋白免疫印迹法检测螯合体1（p62）、微管相关蛋白轻链3（LC3）、酰基辅酶A合成酶长链家族成员4（ACSL4）、谷胱甘肽过氧化物酶4（GPX4）蛋白水平。结果显示，与对照组相比，OGD/R组p62、GPX4蛋白表达显著降低（P<0.05，P<0.01），ACSL4蛋白表达显著升高（P<0.01）；与OGD/R组相比，OV-FUNDC1组LC3、GPX4蛋白表达显著升高（P<0.05，P<0.01），p62和ACSL4蛋白表达显著降低（P<0.05，P<0.01）。第二部分实验，Sal干预后蛋白免疫印迹法检测FUNDC1、p62、LC3、ACSL4、GPX4蛋白水平，JC-1检测线粒体膜电位变化。结果显示，与OGD/R组相比，Sal组FUNDC1、LC3、GPX4蛋白表达显著升高（P<0.01），p62、ACSL4蛋白表达显著降低（P<0.01），线粒体膜电位显著升高（P<0.01）；3MA组线粒体膜电位显著降低（P<0.01）。第三部分实验，沉默FUNDC1后，使用亚铁离子（Fe²⁺）试剂盒检测细胞中Fe²⁺含量；流式细胞术检测细胞活性氧（ROS）水平；MitoTracker Red检测线粒体功能；试剂盒检测细胞三磷酸腺苷（ATP）、谷胱甘肽（GSH）含量；免疫荧光检测ACSL4、GPX4表达；蛋白免疫印迹法检测p62、LC3、ACSL4、GPX4蛋白水平。结果显示，与Si-FUNDC1组相比，Sal组Fe²⁺浓度显著降低（P<0.01）；ROS表达显著降低（P<0.01）；MitoTracker Red检测荧光强度显著升高（P<0.01）；GSH含量显著升高（P<0.01）；ACSL4荧光强度显著下降（P<0.01），GPX4荧光强度显著上升（P<0.01）；LC3、GPX4蛋白表达显著升高（P<0.01），p62、ACSL4蛋白表达显著降低（P<0.01）。该研究揭示FUNDC1可能是缺血再灌注后神经元线粒体自噬与铁死亡的关联靶蛋白，而Sal干预可通过促进非泛素化FUNDC1线粒体自噬途径抑制神经元缺血再灌注后铁死亡。

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