Aug 2026· Toxicology Letters· Vol 424, pp.
113183
· 0 citations· 44 references
Medicine
TL;DR
It is demonstrated that BPA activates the AMPK/mTOR/ULK1 signaling pathway to promote NCOA4‑mediated ferritinophagy, leading to ferroptosis in HT‑22 cells.
Abstract
Bisphenol A (BPA), a widespread environmental endocrine disruptor, is associated with neurodevelopmental disorders and induces oxidative neurotoxicity. Ferroptosis, an iron-dependent cell death driven by lipid peroxidation, has been implicated in toxicant‑induced neuronal injury. However, whether BPA triggers neuronal ferroptosis through autophagy remains unclear. Using HT‑22 hippocampal neuronal cells as an in vitro model, we investigated the role of autophagy‑dependent ferroptosis in BPA neurotoxicity. BPA exposure caused oxidative damage and mitochondrial ultrastructural abnormalities. It also induced ferroptosis‑related changes, including increased malondialdehyde (MDA), prostaglandin endoperoxide synthase 2 (PTGS2) protein expression, and reactive oxygen species (ROS), as well as decreased glutathione (GSH), glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11). These effects were reversed by the ferroptosis inhibitors ferrostatin-1 (Fer-1) and deferoxamine (DFO). Pharmacological inhibition of autophagy with chloroquine (CQ) also reversed BPA-induced GPX4/SLC7A11 downregulation and PTGS2 upregulation. Notably, BPA decreased the expressions of nuclear receptor coactivator 4 (NCOA4) and ferritin heavy chain 1 (FTH1), which was blocked by CQ. Knockdown of NCOA4 attenuated BPA-induced FTH1 and GPX4 loss, and PTGS2 elevation, indicating that NCOA4-mediated ferritinophagy is required for BPA-induced ferroptosis. Mechanistically, BPA activated AMPK/ULK1 axis while inhibiting mTOR; silencing of AMPK or ULK1 partially abrogated BPA-induced autophagy and ferroptosis. Collectively, these findings demonstrate that BPA activates the AMPK/mTOR/ULK1 signaling pathway to promote NCOA4‑mediated ferritinophagy, leading to ferroptosis in HT‑22 cells. This study provides a novel insight into the molecular mechanisms underlying BPA-associated neurotoxicity.
Ferroptosis, an iron-dependent and lipid peroxidation-driven form of regulated cell death, has emerged as a key contributor to neuronal degeneration in protein misfolding disorders. However, how cellular energy metabolism modulates ferroptotic susceptibility in neurodegenerative proteinopathies remains incompletely defined. The present study investigated whether the previously described energy stress-AMP-activated protein kinase (AMPK)-acetyl-CoA carboxylase (ACC) ferroptosis checkpoint operates in disease-relevant neuronal proteinopathy models. Pharmacological and genetic interventions, cell viability assays, lipid peroxidation measurements, western blotting, RNA interference, and behavioral analyses were performed in SH-SY5Y cells, transgenic Caenorhabditis elegans (C. elegans), and 3xTg-AD mice. In SH-SY5Y cells, glucose deprivation, 2-deoxy-D-glucose (2DG), or 5-aminoimidazole-4-carboxamide ribonucleoside reduced ferroptotic cell death induced by erastin or RSL3 and attenuated ferroptosis-associated neuronal injury induced by amyloid precursor protein, α-synuclein A53T, and polyQ74. This protection was associated with AMPK activation, ACC1 phosphorylation, reduced lipid peroxidation, sensitivity to ferroptosis inhibition, glutathione peroxidase 4 restoration, and acyl-CoA synthetase long-chain family member 4 suppression. AMPK inhibition or knockdown weakened the protection conferred by glucose deprivation, while aak-2 RNAi attenuated the protective effects of 2DG in C. elegans, supporting a substantial contribution of AMPK-ACC1-associated signaling to ferroptosis suppression under energy stress. Consistently, 2DG or liproxstatin-1 reduced ferroptosis-associated oxidative damage and improved behavioral or cognitive deficits in transgenic C. elegans and 3xTg-AD mice. These findings extend the established AMPK-ACC ferroptosis checkpoint to neurodegenerative proteinopathy models and support energy stress-associated AMPK-ACC1 signaling as an important metabolic defense mechanism against ferroptosis-related neuronal injury.
Yuanyuan Yong, Yuxin Hu, Qian Feng et al.· International Journal of Mol...· 0 citations
OBJECTIVES
To investigate whether AS-IV alleviates high glucose (HG)-induced podocyte ferroptosis and whether this effect is associated with the GSK3β/Nrf2/GPX4 axis.
METHODS
Differentiated MPC-5 podocytes were exposed to HG (30 mmol/L) with or without AS-IV, the GSK3β inhibitor LY2090314, or the ferroptosis inhibitor Ferrostatin-1 (Fer-1). An osmotic control (mannitol) was included. Cell viability was quantified with the CCK-8 assay. Levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH) and Fe²⁺ were measured. Lipid peroxidation was detected using C11-BODIPY 581/591. Mitochondrial morphology was examined by transmission electron microscopy and protein expression was analyzed by Western blot.
RESULTS
HG exposure induced podocyte injury, characterized by decreased viability, increased oxidative stress (elevated ROS, MDA and lipid ROS), GSH depletion, iron overload and mitochondrial damage. The osmotic control did not reproduce these effects. AS-IV or Fer-1 significantly attenuated the HG-induced damage and lipid peroxidation. At the molecular level, HG downregulated Nephrin, p-GSK3β (Ser9), Nrf2 and GPX4, while upregulating total GSK3β. AS-IV treatment partially reversed these protein expression changes and produced a protective pattern similar to that of LY2090314.
CONCLUSION
AS-IV alleviates HG-induced podocyte injury, possibly by suppressing ferroptosis and this protective effect may involve modulation of the GSK3β/Nrf2/GPX4 axis. The results offer new insights into DN pathogenesis and support AS-IV as a potential therapeutic candidate.
M. Gao, Hong Jiang, Kangya Lei et al.· Pakistan Journal of Pharmace...· 0 citations
Emerging evidence implicates ferroptosis in myocardial ischemic injury. This study aimed to investigate whether empagliflozin (EMP) suppresses ferroptosis in acute myocardial infarction (AMI) via the AMP-activated protein kinase (AMPK)/nuclear factor erythroid 2-related factor 2 (NRF2)/solute carrier family 7 member 11 (SLC7A11) pathway.Hypoxia/reoxygenation (H/R)-injured HL-1 cardiomyocytes were treated with EMP (10, 20, 30 μM). Cell viability, morphology, damage, and apoptosis were assessed by CCK-8, inverted microscopy, lactate dehydrogenase (LDH) release, and flow cytometry. H/R-injured cells were treated with 30 μM EMP and/or ferrostatin-1 (Fer-1), followed by measurements of Fe2+, malondialdehyde (MDA), glutathione (GSH), reactive oxygen species (ROS), superoxide dismutase (SOD), lipid ROS (C11-BODIPY), ferroptosis-related proteins [SLC7A11, glutathione peroxidase 4 (GPX4), phospho-AMPK (p-AMPK), AMPK, NRF2 using Western blot], and NRF2 nuclear translocation (using immunofluorescence). An AMI mouse model was established. Myocardial pathology and infarct size were evaluated, and AMPK/NRF2/SLC7A11 pathway proteins and myocardial Fe2+/MDA/GSH levels were measured.EMP partially reversed H/R-induced cardiomyocyte shrinkage and membrane rupture, reduced viability, elevated LDH, and increased apoptosis. Among cell death inhibitors, Fer-1 exerted maximal protection, indicating ferroptosis may be the predominant death type in H/R-injured HL-1 cardiomyocytes. EMP mirrored the anti-ferroptotic activity of Fer-1. Mechanistically, EMP decreased cytoplasmic NRF2 and the p-AMPK/AMPK ratio while promoting the translocation of NRF2 from the cytoplasm to the nucleus. EMP activated the AMPK/NRF2/SLC7A11 axis to attenuate ferroptosis in AMI mice.In conclusion, EMP suppressed ferroptosis and ameliorated cellular injury in H/R-injured cardiomyocytes by activating the AMPK/NRF2/SLC7A11 pathway, with efficacy confirmed in vivo during AMI.
Introduction As an independent risk factor for atherosclerosis (AS), hyperhomocysteinemia (HHcy) exerts its pathogenic effects primarily through the induction of macrophage ferroptosis. As a selective autophagic process mediated by nuclear receptor coactivator 4 (NCOA4), ferritinophagy directly influences ferroptosis via its regulation of cellular iron balance. However, whether homocysteine (Hcy) regulates macrophage ferroptosis through ferritinophagy remains unclear. Methods Human acute monocytic leukemia (THP-1) cells were differentiated into macrophages and subsequently treated with Hcy. Ferroptosis was assessed by measuring glutathione (GSH) levels, reactive oxygen species (ROS), Fe²⁺ content, and mitochondrial morphology. Protein expression of NCOA4, FTH1, and GPX4 was examined, and GPX4 methylation was evaluated. The involvement of ferritinophagy was further verified using the ferroptosis inhibitor ferrostatin‑1 (Fer‑1) and the inducer Erastin. Activation of the IL‑6/STAT3 signaling pathway was also examined, along with its reciprocal regulation with ferroptosis. Results Hcy treatment promoted ferroptosis in THP‑1 macrophages, as indicated by decreased GSH levels, increased ROS and Fe²⁺ levels, and characteristic mitochondrial morphological changes. Hcy also enhanced GPX4 methylation, resulting in reduced GPX4 expression. Mechanistically, Hcy upregulated NCOA4 and downregulated FTH1, suggesting activation of NCOA4‑mediated ferritinophagy; these effects were reversed by Fer‑1 and augmented by Erastin. In addition, Hcy activated the IL‑6/STAT3 pathway, and its crosstalk with ferroptosis was confirmed by the reciprocal modulation with Fer‑1 and Erastin. Discussion Collectively, our study indicates that Hcy promotes ferroptosis in THP-1 macrophages through NCOA4-mediated ferritinophagy, and the IL-6/STAT3 signaling pathway plays a key role in this process. Therefore, targeting Hcy may represent a potential treatment strategy for AS.
Oxidative stress is increasingly recognized as a critical pathogenic factor in ovarian granulosa cell dysfunction and the progression of female reproductive disorders. Accumulating evidence suggests that environmental toxicants contribute to ovarian pathogenesis. For example, aflatoxin B1 (AFB1), a widespread foodborne mycotoxin, is a potent inducer of oxidative stress. After metabolic activation, AFB1 generates reactive oxygen species, causing lipid, protein, and mitochondrial dysfunction. AFB1 also triggers ferroptotic cell death through iron accumulation, glutathione peroxidase 4 downregulation, and lipid peroxidation. Mangiferin (MA), a natural polyphenolic compound with potent antioxidant and cytoprotective effects, has been reported to modulate redox-sensitive signaling pathways. However, whether MA protects human ovarian granulosa cells against oxidative stress and ferroptotic cascades induced by AFB1 exposure remains unclear. In this study, we examined the mechanisms by which MA attenuates AFB1-induced oxidative stress and ferroptosis in granulosa cells through the activation of Nrf2 signaling and mitochondrial biogenesis. These findings highlight the potential of MA as a nutraceutical candidate for alleviating AFB1-induced granulosa cell toxicity.
Chi-Wai Ruan, Hsin-Yi Tsai, Chih-Hsin Hung et al.· Food and Chemical Toxicology· 0 citations
Ferroptosis is an iron-dependent form of regulated cell death driven by lipid peroxidation and glutathione (GSH) depletion and represents a therapeutic vulnerability in hepatocellular carcinoma (HCC). While canonical ferroptosis regulation centers on cystine uptake and GPX4-mediated GSH utilization, the endogenous metabolic pathways governing ferroptosis sensitivity in liver tumors remain incompletely understood. Here, using a metabolic-scale CRISPR activation screen integrated with transcriptomic and metabolomic analyses, we identify L-2-hydroxyglutarate dehydrogenase (L2HGDH) as a potent antagonist of ferroptosis in HCC. We demonstrate that L2HGDH is frequently suppressed in liver tumors, leading to pathological accumulation of its substrate L-2-hydroxyglutarate (L2HG). Elevated L2HG sensitizes HCC cells to ferroptosis both in vitro and in vivo. Mechanistically, L2HG acts as a metabolic-epigenetic regulator that inhibits 2-oxoglutarate-dependent dioxygenases, induces histone hypermethylation, and remodels chromatin accessibility to activate an ATF3-dependent transcriptional program. This program induces the glutathione-degrading enzyme CHAC1, thereby accelerating GSH degradation to 5-oxoproline and disrupting redox homeostasis. Notably, L2HG-induced ferroptosis occurs independently of impaired cystine uptake, transsulfuration pathway activity, or increased GPX4-mediated GSH utilization, revealing a non-canonical ferroptosis mechanism driven by enhanced GSH catabolism. Consistent with these findings, genetic targeting of L2HGDH suppresses tumor growth, elevates L2HG levels, enhances GSH degradation, and promotes ferroptosis in HCC xenograft models. Collectively, our study identifies the L2HGDH-L2HG axis as a previously unrecognized metabolic checkpoint controlling ferroptosis sensitivity in liver cancer and uncovers glutathione degradation as a therapeutically exploitable vulnerability for ferroptosis-based treatment strategies in HCC.
Caixia Xi, Junfeng Pang, Mohammad Oliaeimotlagh et al.· Cell Death and Disease· 1 citation
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