Liver ischemia-reperfusion injury (IRI) is a sterile inflammatory process that contributes significantly to graft rejection following liver transplantation. Although SET domain bifurcated histone lysine methyltransferase 1 (SETDB1) is known to preserve genomic stability and restrain inflammation under oxidative stress, its immunoregulatory function in myeloid cells during liver IRI has not been elucidated. This study aimed to investigate the role and mechanism of SETDB1 in regulating macrophage-driven inflammatory responses in liver IRI. Myeloid-specific SETDB1 knockout (SETDB1 cKO) mice exhibited exacerbated liver injury, increased infiltration of pro-inflammatory macrophages and neutrophils, and amplified inflammatory responses compared with SETDB1fl/fl controls. Depletion of macrophages alleviated liver damage, reduced neutrophil infiltration and hepatocyte apoptosis, and eliminated the excessive injury observed in SETDB1 cKO mice. Mechanistically, SETDB1 suppressed the expression of purinergic receptor P2X7 (P2RX7). Pharmacological inhibition of P2RX7 with oxidized adenosine triphosphate significantly attenuated liver injury and macrophage infiltration in SETDB1 cKO mice. In vitro assays confirmed that SETDB1 inhibited the P2RX7/Caspase-1/Gasdermin D (GSDMD) pathway in macrophages, thereby limiting pyroptosis and inflammation. This study identifies SETDB1 as a previously unrecognized regulator of macrophage pyroptosis during liver IRI. By linking epigenetic regulation to suppression of the P2RX7/Caspase-1/GSDMD pathway, our findings provide novel mechanistic insight into how SETDB1 protects against sterile liver inflammation. SETDB1 plays a pivotal role in protecting the liver from IRI by restraining macrophage-mediated pyroptosis and inflammation. These findings suggest that targeting the SETDB1/P2RX7/Caspase-1/GSDMD axis may represent a promising therapeutic strategy for mitigating liver IRI and improving transplant outcomes. Antioxid. Redox Signal. 45, 133-148.

To determine whether physalin A (PA) safeguards the outer blood-retinal barrier under diabetic stress by engaging nuclear factor erythroid 2-related factor 2 (Nrf2) to restore redox balance and restrain ferroptosis in human retinal pigment epithelial (hRPE) cells and C57BLKS/J Iar -+Leprdb/+Leprdb mice. In high-glucose challenged hRPE cells, PA dose-dependently preserved viability, maintained claudin-1/occludin/zonula occludens-1 abundance and membrane localization, and reversed ferroptosis hallmarks (restored solute carrier family 7 member 11 [SLC7A11], SLC3A2, and glutathione peroxidase 4; reduced ferrous iron [Fe2+] overload and lipid peroxidation). PA restored glutathione levels, reduced malondialdehyde (MDA), and enhanced the antioxidant defense pathway mediated by Nrf2, including upregulation of heme oxygenase 1, NAD(P)H quinone dehydrogenase 1, and superoxide dismutase 2. Silencing Nrf2 abolished the effects of PA on barrier integrity and ferroptosis suppression, with rebounds in reactive oxygen species, MDA, Fe2+, and tight junction loss. In db/db mice treated systemically for 20 weeks, PA reduced Evans Blue leakage, increased retinal thickness, restored RPE tight junction proteins, and normalized mitochondrial architecture by transmission electron microscopy. PA rebalanced mitochondrial dynamics (dynamin 1-like, optic atrophy 1, fission 1, mitofusin 1, FUN14 domain containing 1), increased retinal mitochondrial DNA copy number, and partially stabilized glycemia and weight. PA restores redox tone, restrains ferroptosis, and preserves junctional integrity to protect the diabetic retina, with Nrf2 being indispensable for these benefits. These findings position PA as a promising adjunctive candidate for early diabetic retinopathy and support Nrf2-centered strategies to reinforce the outer blood-retinal barrier. Antioxid. Redox Signal. 45, 5-27.

Cerebral ischemia/reperfusion (I/R) injury represents a significant challenge to recanalization therapy for ischemic stroke and is critically influenced by microglial polarization. Although inhibition of Rho-associated protein kinase (ROCK) has been shown to mitigate cerebral I/R injury and associated neuroinflammation, its specific effect on the balance between M1 and M2 (anti-inflammatory) microglial polarization remains incompletely understood. This study aimed to elucidate the role and underlying mechanism of ROCK inhibition in regulating M1 and M2 microglial polarization, using the classical antidepressant fluoxetine as a positive control. ROCK inhibitor fasudil and positive control fluoxetine effectively alleviated cerebral I/R injury and facilitated a shift in microglial polarization from the M1 to the M2 phenotype, both invivo and in vitro. In the hippocampal tissues of cerebral I/R mice exposed to lipopolysaccharide, we observed an upregulation of thioredoxin-interacting protein (TXNIP) and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX2). ROCK2 knockdown promoted the M2 microglial polarization, suppressed the expression of NOX2 and TXNIP, and inhibited the activation of NF-κB P65 in mouse hippocampal tissue. Notably, pharmacological inhibition of NF-κB reduced the expression of NOX2 and TXNIP, as well as the production of reactive oxygen species (ROS), in microglia subjected to oxygen-glucose deprivation/reoxygenation. Correspondingly, inhibition of NOX2 also decreased TXNIP expression.Conclusion and Innovation:ROCK inhibition promotes a shift in microglial polarization from the M1 to the M2 subtype by suppressing the NF-κB/NOX2/ROS/TXNIP signaling pathway. This study provides the first evidence demonstrating the mechanism by which ROCK inhibition drives microglial polarization toward the M2 phenotype. Antioxid. Redox Signal. 45, 149-168.

The activation of microglia triggers an inflammatory response, which is frequently associated with an imbalance of iron metabolism. This study aimed to determine whether inflammation-associated iron dyshomeostasis contributes to impaired poststroke recovery and to explore the underlying mechanisms. Ferroportin 1 (FPN1) deficiency in neurons and glial cells delayed sensorimotor function recovery following cerebral ischemia. FPN1 deficiency was associated with aggravated neuronal injury, enhanced apoptosis- and necroptosis-associated signaling, impaired myelin- and synapse-related repair, and reduced dendritic spine density in the ischemic cortex. Histological analyses, including hematoxylin and eosin staining and Nissl staining, further supported more severe peri-infarct pathological damage in Fpn1Nestin-CKO mice. In addition, increased IgG extravasation indicated aggravated blood-brain barrier (BBB) disruption and secondary neurovascular injury after stroke. These pathological changes were accompanied by increased iron accumulation in the ischemic cortex and altered expression of iron metabolism-related molecules. Elevated inflammatory cytokine expression and increased hepcidin levels were associated with disrupted brain iron homeostasis in Fpn1Nestin-CKO mice. Inhibition of JAK-STAT signaling with AG490 reduced p-STAT3 and hepcidin levels and was associated with modulation of iron-related and repair-associated responses, with more pronounced effects observed in Fpn1-deficient mice. These findings highlight a close association between inflammatory signaling, BBB dysfunction, and iron dyshomeostasis during poststroke recovery. Our results suggest that delayed sensorimotor recovery in mice with neuronal and glial FPN1 deficiency may be linked to inflammation-associated BBB disruption and subsequent iron accumulation in the ischemic brain. Antioxid. Redox Signal. 00, 000-000.

This study aimed to delineate a novel mechanistic axis linking hyperglycemia-driven glycolytic reprogramming to ferroptotic death in lens epithelial cells (LECs) and to determine its therapeutic significance in diabetic cataract (DC). Specifically, we sought to define the integration of metabolic, epigenetic (histone lactylation), and post-translational (fucosylation) pathways in DC pathogenesis. Under hyperglycemic conditions, LECs exhibited robust glycolytic activation and lactate accumulation. This metabolic shift drove selective histone H3K18 lactylation at the promoter of theTSTA3 gene, leading to its increased transcription. The upregulated TSTA3 protein then promoted the core fucosylation of the NF-κB p50 subunit, which facilitated its nuclear translocation. Inside the nucleus, p50 transcriptionally activatedNOX1, resulting in excessive reactive oxygen species (ROS) production and subsequent ferroptotic cell death. Critically, both pharmacological inhibition of glycolysis and genetic silencing ofTSTA3 effectively attenuated oxidative stress, restored redox balance, and ameliorated cataract severity in a diabetic rat model. This work identifies a previously unrecognized pathogenic cascade-the glycolysis-histone lactylation-fucosylation-ferroptosis axis-that directly links metabolic flux to epigenetic and signaling control in DC. By positioning TSTA3 as a central, druggable node within this axis, our study redefines cataract pathogenesis beyond simple oxidative damage, integrating multiple layers of cellular regulation. The glycolysis-histone lactylation-TSTA3-fucosylation-NOX1-ferroptosis axis is a critical driver of LEC death in diabetic cataract. Targeting this newly defined pathway, particularly the TSTA3 node, offers novel opportunities for mechanism-based therapeutic interventions and biomarker development, with potential implications for other complications of metabolic disease.

Doxorubicin (DOX) remains a cornerstone of cancer therapy but is limited by dose-dependent cardiotoxicity with inadequate protective strategies. Nerve injury-induced protein 1 (NINJ1), a regulator of inflammation and cell death, has not been explored in this context. We sought to define the role of NINJ1 in DOX-induced cardiotoxicity and evaluate its translational potential. Using complementary genetic, pharmacologic, and transcriptomic approaches, we demonstrate that NINJ1 is markedly upregulated in DOX-treated murine hearts and cardiomyocytes. Cardiomyocyte-specific NINJ1 deletion confers robust protection against cardiac dysfunction, oxidative stress, and apoptosis, whereas NINJ1 overexpression exacerbates injury. Mechanistically, NINJ1 suppresses AMP-activated protein kinase (AMPK) activation, promoting ubiquitin-mediated degradation of hypoxia-inducible factor-1α (HIF-1α), thereby impairing antioxidant gene programs. Multilevel evidence, including RNA sequencing, pathway enrichment, and gain- and loss-of-function models, establishes the NINJ1-AMPK-HIF-1α axis as a central regulator of redox homeostasis. Pharmacologic inhibition of NINJ1 with phenyl-β-D-glucopyranoside attenuates cardiac injury invivo and in vitro without compromising DOX antitumor efficacy, supporting pathway specificity and therapeutic feasibility. This study identifies NINJ1 as a previously unrecognized driver of anthracycline cardiotoxicity and uncovers a novel signaling axis linking membrane injury signaling to metabolic control of HIF-1α stability. NINJ1 promotes DOX-induced cardiotoxicity by destabilizing HIF-1α via AMPK inhibition. Targeting NINJ1 represents a promising cardioprotective strategy. Therapeutic inhibition of NINJ1 protects the heart while preserving anticancer efficacy, offering a potential strategy to enhance the safety of anthracycline-based chemotherapy and improve outcomes in cancer patients. Antioxid. Redox Signal. 00, 000-000.

Sjögren's syndrome (SS) is a systemic autoimmune disorder characterized by chronic inflammation, oxidative stress, and progressive salivary gland dysfunction. Current therapies remain limited in efficacy. This study explored the regulatory effect of exosome (Exo)-transported miR-23b-3p on the IκB kinase alpha (IKKα)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling axis in SS. Key SS-related microRNAs (miRNAs) were identified by integrating weighted gene co-expression network analysis with machine-learning-based transcriptomic profiling. Target genes and functional pathways were analyzed by bioinformatics methods. The direct binding between miR-23b and IKKα (CHUK) was validated by a dual-luciferase reporter assay. An in vitro SS cell model was established to examine the effects of miR-23b-3p on the IKKα/NF-κB pathway, oxidative stress, inflammation, and apoptosis. miR-23b-3p was loaded into salivary gland epithelial cell-derived Exos (SGEC-Exos) via electroporation. In vitro coculture experiments assessed reactive oxygen species (ROS) levels, inflammatory cytokines, Treg/Th17 balance, and cell apoptosis. In vivo effects were evaluated in NOD/Ltj mice by measuring salivary flow rate, histopathology, and expression of salivary-function-related proteins AQP5 and GPER. SGEC-Exos@miR-23b-3p significantly suppressed IKKα expression and NF-κB activation, reduced ROS production, and modulated immune responses by restoring the Treg/Th17 balance. It also inhibited apoptosis by decreasing Bax and caspase-3 expression and increasing Bcl-2 levels. These effects were partially reversed by reactivation of the IKKα/NF-κB pathway. In NOD/Ltj mice, SGEC-Exos@miR-23b-3p improved salivary flow, alleviated glandular pathology, and upregulated AQP5 and GPER expression. SGEC-Exos@miR-23b-3p offers a translational approach to address oxidative stress, immune imbalance, and glandular injury in SS, highlighting the potential of Exo-based miRNA therapy. Antioxid. Redox Signal. 00, 000-000.

Sleeve gastrectomy (SG) improves obesity-associated type 2 diabetes mellitus (T2DM) beyond mere weight loss. We investigated whether SG enhances systemic metabolic homeostasis by suppressing the Ghrelin-growth hormone secretagogue receptor (GHSR) axis, remodeling hypothalamic pro-opiomelanocortin (POMC) neuronal activity, and reprogramming CD4+ T cell immunometabolism. Using a diet-induced T2DM mouse model undergoing SG or Sham surgery, we integrated bulk/single-cell RNA sequencing and metabolomics to evaluate systemic neuro-immune-metabolic alterations. Functional assays validated Ghrelin's effects on CD4+ T cell metabolism and differentiation, alongside assessments of hepatic/pancreatic function and hypothalamic neuronal activity. SG globally remodeled peripheral immunity, expanding Tregs while reducing pro-inflammatory Th17 cells. scRNA-seq and metabolomic profiling revealed that CD4+ T cells shifted metabolically from glycolysis toward oxidative phosphorylation, matching increased tricarboxylic acid cycle intermediates. Functionally, Ghrelin-GHSR signaling promoted CD4+ T cell glycolysis, mitochondrial damage, and Th17 skewing; GHSR antagonism successfully reversed these detrimental effects. Systemically, SG reduced hyperglycemia and hepatic lipidosis, restored islet α/β-cell balance, activated anorexigenic POMC neurons, and suppressed AgRP neurons. SG alleviates T2DM through coordinated suppression of the Ghrelin-GHSR axis, bridging central appetite regulation with peripheral immunometabolic reprogramming. By shifting CD4+ T cells toward oxidative metabolism and restoring the Treg/Th17 balance, SG drives systemic metabolic recovery, providing critical molecular insights into the neuro-immune mechanisms of metabolic surgery. Antioxid. Redox Signal. 00, 000-000.

Acute central nervous system (CNS) injuries impose a significant global burden. Microsurgical decompression effectively stabilizes primary anatomy. However, it often fails to stop the complex biochemical cascades of secondary neurodegeneration. There is a critical need to bridge the gap between anatomical preservation and functional recovery. Strong preclinical evidence indicates that delayed bioenergetic failure within the injury microenvironment heavily dictates long-term outcomes. We synthesize the ARFE (autophagy-reactive oxygen species-ferroptosis-edema) axis as a mechanistic framework delineating the pathological continuum from subcellular failure to macroscopic tissue edema. In this irreversible cascade, adenosine triphosphate depletion blocks autophagic flux, forcing ferritinophagy-driven iron release and lipid peroxidation, while succinate accumulation locks microglia in metabolic collapse. A translational gap persists because mechanical hematoma evacuation does not inherently reverse the metabolic cascades driving secondary injury. Current single-target modalities fail because they do not account for the evolving metabolic microenvironment, leading to unchecked inflammation and cell death despite successful surgical intervention. We propose a paradigm shift from single-target modalities to "spatiotemporal metabolic engineering." This strategy synchronizes interventions with metabolic logic. Hyperacute treatments focus on redox containment to neutralize iron. Acute phases prioritize immune-metabolic reprogramming for inflammation. Finally, subacute stages aim for bioenergetic reconstruction to support axonal regrowth. Antioxid. Redox Signal. 00, 000-000.