Age-related muscle atrophy is closely associated with mitochondrial dysfunction and ferroptosis. This study established a D-gal-induced sarcopenia model in aged mice and a C2C12/GM17940 cell myotube senescence model, with young/control, old/D-gal, and formononetin (FMN) intervention groups. After shSIRT1 transfection and mitochondrial-targeted antioxidant Mito-C intervention, the effects and mechanism of FMN were detected by measuring mouse phenotypic indicators (lean mass, hindlimb muscle mass, grip strength) and cell indicators (viability, mitochondrial membrane potential, ROS, ATP, ferroptosis-related proteins). Results showed that FMN improved lean mass, grip strength, mitochondrial membrane potential, and ATP production, while reducing ROS and ferroptosis by regulating ACSL4, GPX4, and SLC7A11. Mechanistically, FMN exerted protective effects via the SIRT1/PGC-1α pathway, which was partially attenuated by SIRT1 knockdown or Mito-C. Collectively, FMN alleviates age-related sarcopenia by targeting mitochondrial function and ferroptosis, providing potential targets for sarcopenia treatment.
Red blood cell (RBC)-based therapeutic enzyme delivery systems require cell sources that can support future standardized large-scale production, with the prerequisite that engineering modifications preserve the cells' inherent biocompatibility and long-circulating potential. In this study, we first demonstrated proof-of-principle by successfully engineering asparaginase (ASPG)-loaded erythroid cells in the human erythroid progenitor cell line (HUDEP-2), where efficient ASPG expression and intact enzymatic activity were confirmed. To facilitate clinical translation, the validated strategy was further applied to human induced pluripotent stem cells (iPSCs), and the differentiated products were systematically characterized. It is worth noting that the key membrane markers of iPSC-derived ASPG-loaded erythroid cells (i-ASPG-R), including CD47 and CD55, were comparable to those of human RBCs (hRBCs). The results of Annexin V staining indicated a healthy cell status. More importantly, these cells displayed ASPG activity equivalent to that of HUDEP-2-derived counterparts. Moreover, the expression of ASPG did not affect enucleation and the composition of globin. In vitro function assays showed that compared with the control group, the proliferation of CCRF-CEM leukemia cells was inhibited by 48% after 24 h of co-culture with i-ASPG-R. In summary, this study established an iPSC-derived platform for generating ASPG-loaded erythroid cells with favorable carrier properties and evident anti-leukemic activity in vitro, laying a conceptual foundation for future cell therapy manufacturing via optimized terminal maturation.
BACKGROUND:Type 2 diabetes mellitus (T2DM) is one of the most common chronic diseases, whose prevalence is increasing worldwide. There is a great demand for an effective cure that attains a normal glucose level. The differentiation of mesenchymal stem cells into insulin-producing cells (IPCs) for pancreatic regeneration represents a promising anti-diabetic therapeutic approach. However, the pathogenic milieu of T2DM may affect their differentiation potential. AIM:This study aimed to investigate the impact of the T2DM milieu on the pancreatic differentiation potential of human adipose-derived stem cells (ASCs) in vitro. METHODS:We isolated and cultured ASCs from T2DM (dASCs) and non-diabetic (nASCs) subjects, then we differentiated the cells into IPCs. We evaluated the differentiation potential using dithizone staining, immunofluorescence staining, and gene expression analysis. We examined the functionality of generated cells via glucose challenge assay. RESULTS:Our results revealed that both dASCS and nASCs could be committed to an early endocrine-like phenotype with comparable potential. However, the glucose-stimulated insulin secretion of dASCs-derived IPCs was significantly less than that derived from nASCs. CONCLUSION:T2DM does not affect the pancreatic commitment potential of dASCs. However, ASCs-derived from diabetic patients displayed a reduced capacity to acquire glucose-responsive endocrine characteristics following pancreatic differentiation, exhibiting a limited β-like functionality.
We demonstrate that SARS-CoV-2 exploits tunneling nanotubes (TNTs) as actin-based conduits for intercellular transmission in epithelial cells. TNT formation was markedly enhanced in the Delta variant compared with the ancestral Wuhan-Hu-1 and Omicron strains, consistent with their relative transmissibility. The Delta variant also promoted extensive filopodia and branched TNT networks, facilitating efficient cell-to-cell dissemination. Confocal and ultrastructural analyses revealed active transfer of mitochondria and lysosomes through these nanotubes, indicating that SARS-CoV-2 co-opts host organelle trafficking to support replication and persistence. Pharmacological inhibition of Cdc42-dependent actin remodeling or CaMKII signaling disrupted TNT formation. Collectively, these findings identify TNTs as a crucial pathway for variant-dependent SARS-CoV-2 dissemination in epithelial cells and reveal host cytoskeletal dynamics as a tractable antiviral target for limiting viral spread.
Protein S-palmitoylation is a reversible lipid post-translational modification that dynamically controls protein localization, trafficking, receptor microdomain organization, autophagy and metabolic signalling. In osteoarticular tissues, this modification provides a plausible biochemical mechanism through which osteoclasts, osteoblast-lineage cells, osteocytes, chondrocytes, synoviocytes and skeletal-muscle cells integrate inflammatory, mechanical and metabolic cues. This review synthesizes current evidence linking S-palmitoylation and depalmitoylation to osteoarticular cell signalling and metabolic adaptation. We focus on experimentally supported mechanisms, including ZDHHC-mediated palmitoylation; depalmitoylation by APT, ABHD and PPT enzymes; palmitoylation-sensitive osteoclast differentiation; inflammatory priming of macrophage-derived osteoclast precursors; BMP/SMAD and organelle-contact-site signalling in osteoblast-lineage cells; ZDHHC11-dependent chondrocyte protection; ZDHHC4-mediated palmitoylation of CCDC50 followed by autophagic clearance of MAP2K4/MKK4; inflammasome regulation; CD36-associated lipid uptake; and palmitoylation-dependent transporter localization and turnover. We distinguish direct skeletal and joint evidence from mechanistic analogues derived from immune, neural, metabolic and cancer systems. We also clarify the distinction between reversible cysteine S-palmitoylation, broader S-acylation and other lipid modifications. Major barriers include incomplete definition of enzyme and substrate relationships, false-positive risk in palmitoyl-proteomic workflows, limited temporal resolution, insufficient validation in primary human tissues, uncertain tissue specificity and the off-target effects of non-selective palmitoylation inhibitors. A cell-biochemistry-centred view of the dynamic balance between palmitoylation and depalmitoylation may clarify how lipid modification regulates osteoarticular cell function and may help prioritize experimentally testable, substrate-specific mechanisms for future validation in osteoarthritis, osteoporosis and inflammatory joint disease.
Hypertension is a major risk factor for cardiovascular disease and alters the mechanical microenvironment of the vascular wall and target organs. Key mechanical disturbances include disturbed shear stress, excessive circumferential stretch, elevated hydrostatic pressure, and increased extracellular matrix stiffness. These forces are sensed by multiple classes of mechanosensors, including mechanosensitive ion channels, integrin-based adhesion complexes, G protein-coupled receptors, primary cilia, baroreceptor afferents, and nuclear mechanosensing structures. Activation of these sensors engages interconnected Ca²⁺-dependent, RhoA/Rho-associated protein kinase, mitogen-activated protein kinase, phosphoinositide 3-kinase/Akt/endothelial nitric oxide synthase, Yes-associated protein/transcriptional coactivator with PDZ-binding motif, redox-sensitive, inflammatory, and mechano-epigenetic pathways. This review summarizes how altered mechanotransduction may contribute to endothelial dysfunction, vascular smooth muscle cell phenotypic switching, vascular remodeling, and cardiac and renal target-organ injury in hypertension. Mechanotransduction is unlikely to represent a universal initiating cause of hypertension. Rather, it should be viewed as a context-dependent set of adaptive and maladaptive responses that varies according to cell type, vascular bed, mechanical stimulus, disease stage, and experimental model. Physiological mechanotransduction helps maintain vascular homeostasis and baroreflex function, whereas sustained pathological loading may amplify oxidative stress, inflammation, fibrosis, and vascular stiffening. However, most direct mechanistic evidence derives from cultured cells and animal models, and human genetic, tissue, biomarker, and interventional data remain limited. Conventional antihypertensive therapy therefore remains the foundation of clinical management, whereas direct targeting of mechanosensors or downstream mechanotransduction pathways remains experimental and requires stronger human validation.
Qi Huang Granules (QHG), a traditional Chinese medicine (TCM) formulation, have been applied clinically for over two decades to treat dry age-related macular degeneration (AMD) and associated fundus lesions. Although its retinoprotective effects have been documented, the associated underlying mechanisms are largely unexplored. The present work focused on investigating the therapeutic efficacy of QHG in sodium iodate (NaIO₃)-induced retinal damage, with a particular focus on how the formula regulated the interaction between inflammatory responses and imbalances in mitochondrial dynamics. UPLC-HRMS, network pharmacology, and molecular docking analyses were integrated for identifying mitochondria-associated bioactive constituents and potential targets of QHG. In the experimental model, retinal injury was induced in rats through tail vein injection of NaIO3. Retinal morphological and ultrastructural changes were assessed by HE staining and transmission electron microscopy. To assess mitochondrial function, mitochondrial membrane potential, mtROS levels, and mtDNA integrity were measured. Additionally, ELISA was performed to quantify IL-1β and IL-18 levels. Key marker levels, including p-DRP1, DRP1, OPA1, MFN2, and NLRP3, were determined by Western blotting, IHC, and qRT-PCR. DNM1L (encoding DRP1) and MFN2 were identified as the primary targets of QHG through UPLC-HRMS and computational analyses. In vivo experimental results showed that treatment with QHG alleviated morphological alterations and ultrastructural damage to the retina and mitochondria in model rats. Furthermore, QHG treatment increased mitochondrial membrane potential, reduced mtROS levels, mitigated mtDNA damage, and reduced IL-1β and IL-18 contents. Mechanistically, QHG downregulated p-DRP1, DRP1, and NLRP3, while upregulating MFN2 and OPA1. Collectively, these findings demonstrate that QHG can ameliorate retinal morphology and ultrastructural damage in mitochondria, such as cellular mitochondria, improve mitochondrial function, and attenuate retinal inflammation, thereby exerting a protective effect against NaIO₃-induced retinal injury. The underlying mechanism may involve the mitochondrial dynamics-NLRP3 pathway.
Intracerebral hemorrhage (ICH)-induced secondary brain injury (SBI) is a critical contributor to poor neurological outcomes. In this study, we aimed to identify potential small‑molecule therapeutic agents and systematically validate their neuroprotective efficacy. Bioinformatics analysis was performed to screen ICH‑related hub genes, and Connectivity Map (CMap) was used to predict candidate compounds, followed by molecular docking to verify their binding affinity. The therapeutic effect of TPCA‑1 was further evaluated in a mouse ICH model using neurological scoring, cerebral edema assessment, qPCR, Western blot, and ELISA. Bioinformatics analysis indicated that the chemokine signaling pathway is critically involved in ICH pathogenesis. Molecular docking revealed that TPCA‑1 displayed the strongest binding affinity for CXCR4 among all predicted compounds. In vivo, 20mg/kg TPCA‑1 significantly ameliorated neurological deficits and reduced cerebral edema in ICH mice. Mechanistically, TPCA-1 interacted with CXCR4, as supported by molecular docking, MST, and CETSA assays, and suppressed downstream inflammatory signaling involving NF-κB and AP-1, thereby decreasing the production of pro‑inflammatory cytokines including TNF‑α, IL‑1β, and IL‑6. Importantly, Co-treatment with the CXCR4 inhibitor AMD3100 did not produce additional inhibitory effects. Collectively, TPCA‑1 exerts neuroprotective effects against ICH‑induced SBI by targeting CXCR4, which further inhibits the activation of NF‑κB and AP‑1 and ultimately alleviates neuroinflammation.
BACKGROUND:Glioblastoma (GBM) is the most aggressive and lethal primary brain tumor, characterized by poor prognosis and resistance to current therapies. Boron neutron capture therapy (BNCT) is a tumor-selective particle radiation therapy in which high-linear energy transfer (LET) α-particles and recoil 7Li nuclei are generated through the capture reaction between 10B and thermal neutrons. Although BNCT has been shown to prolong survival in GBM, its effects on global metabolomic alterations of the whole brain, particularly in relation to the tumor microenvironment (TME), remain incompletely understood. METHODS:C57BL/6 mice orthotopically implanted with GL261 glioma cells were administered p-boronophenylalanine (BPA) at a dose of 500 mg/kg, followed by thermal neutron irradiation. Brain tissues were analyzed using conventional histopathological methods, including immunohistochemistry, as well as matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). RESULTS:Histopathological analysis demonstrated marked regression of the tumor mass following BNCT. Non-targeted MALDI-MSI enabled the extraction and spatial visualization of tumor-associated metabolites, among which UDP-GlcNAc emerged as one of the most prominent oncometabolites in the present analysis. In addition, ADP signals were predominantly colocalized within the corpus callosum of tumor-bearing brains. A metabolic gradient shift from the tumor-implanted hemisphere to the contralateral side was observed, potentially reflecting tumor cell migration. Uniform Manifold Approximation and Projection (UMAP) analysis facilitated the visualization of extracellular metabolites in the ventricular regions, and elevated GDP signals were associated with hydrocephalus. Notably, the adenylate energy charge of the brain was significantly increased after BNCT, followed by a gradual decline over time, whereas it was reduced within the tumor region after treatment. CONCLUSIONS:Non-targeted MALDI-MSI of brains from a murine glioma model successfully visualized purinergic and pyrimidinergic metabolites that may reflect tumor dynamics, BNCT efficacy, and metabolic responses of the normal brain to treatment. These findings suggest the potential utility of this approach for identifying diagnostic and prognostic biomarkers in glioblastoma.
INTRODUCTION:Intestinal ischemia-reperfusion (I/R) injury is a critical clinical condition involving oxidative stress, inflammation, and immune dysregulation, often leading to significant morbidity and mortality. This study investigates the protective mechanisms of Heme oxygenase-1 (Hmox1) against I/R-induced intestinal damage, focusing on its regulation of Th17/Treg cell balance via Lipocalin 2 (LCN2). MATERIAL AND METHODS:Male wild-type mice and Hmox1-deficient mice were used to induce an intestinal I/R injury model. Histological analysis, ELISA, immunohistochemistry, immunofluorescence, flow cytometry, RT-qPCR, Western blotting, and RNA sequencing were used to evaluate intestinal injury, cytokine levels, and immune cell profiles. Transcriptomic analysis was conducted to identify differentially expressed genes and pathways involved in the immune response. Naive CD4 + T cells were isolated for in vitro differentiation experiments to analyze the role of LCN2 in Th17 and Treg differentiation under Hmox1-deficient conditions. RESULTS:Hmox1 deficiency was associated with more severe intestinal mucosal injury, higher IL-6 and TNFα levels, enhanced Th17-associated responses, and reduced Treg-associated signals after I/R. Transcriptomic analysis indicated enrichment of IL-17, JAK-STAT, and TNFα signaling pathways. LCN2 expression increased after I/R and was further elevated in Hmox1-deficient I/R samples. In vitro, LCN2 knockdown enhanced Th17 polarization and further reduced Treg differentiation under oxidative/inflammatory stimulation, suggesting that LCN2 may participate in a compensatory immune-regulatory response. CONCLUSION:These findings underline the crucial role of Hmox1 in preserving intestinal immune homeostasis during I/R injury and identify LCN2 as a candidate compensatory regulator of Th17/Treg balance under Hmox1-deficient conditions. LCN2 may therefore represent a potential target for non-Hmox1-dependent strategies to ameliorate intestinal I/R injury.
Chloroquine (CQ) is known for anti-inflammatory and autophagy-inhibitory properties; however, its impact on endotoxin-induced macrophage response remains unclear. This study aims to investigate how CQ acts as a signalling modulator in the oxidative stress-autophagy-apoptosis axis in presence of LPS and how the modulation of TLR4 and NLRP3 affects that axis in murine splenic macrophages. The isolated macrophages were pretreated with TLR4 Ab and NLRP3 inhibitor along with CQ for 30 min and further for an additional 60 min after LPS addition. From the in vitro stimulated cells several functional and cellular parameters along with expression of key signalling protein markers were analysed. Result shows that CQ administration in presence of LPS enhances the intracellular ROS which may contribute to alteration of AMPK-ULK1 signalling. The downregulation of pAMPK-pULK1 confirms that CQ + LPS is associated with alterations in upstream autophagy-regulatory signalling markers and consistent with impaired autophagy-associated processing. Additionally, the accumulation of p62 along with LC3-II cautiously acts as a critical indicator of autophagosome degradation inhibition in response to CQ and LPS. The NLRP3 activation, p62 accumulation further corroborate with the Bax and cleaved caspase 3 overexpression in CQ and LPS administered cells which confirm the apoptosis induction. Restoration of pAMPK-pULK1 reduces the mTOR level and restarts the autophagic phenomenon in the case of dual inhibition of TLR4 and NLRP3 in presence of CQ, LPS. The blocking of TLR4 and NLRP3 is able to mitigate the inflammation by restarting the autophagy and limiting apoptosis in murine splenic macrophages.
Diabetes mellitus (DM) is a global health concern characterized by chronic hyperglycemia, which drives severe microvascular and macrovascular complications. The reciprocal interaction between hyperglycemia and reactive oxygen species (ROS)-induced oxidative stress represents a fundamental mechanism in diabetic pathogenesis. As recent studies identify novel intracellular pathways and highlight the critical role of mitochondria, re-examining the interplay between oxidative stress and intracellular organelles is essential. Unlike conventional models that view oxidative stress merely as a passive downstream consequence of cellular damage, this review introduces a distinct, integrative perspective. We reframe the hyperglycemia-oxidative stress axis as a highly coordinated, organelle-driven signaling network that physically bridges metabolic dysfunction with sterile inflammation. By synthesizing classical metabolic shunts (mitochondrial dysfunction, the polyol pathway, advanced glycation end products (AGEs), protein kinase C (PKC) activation, and the hexosamine pathway) with modern inflammatory signaling cascades, we delineate how hyperglycemia amplifies ROS production. This amplification triggers β-cell dysfunction, insulin resistance, and progressive tissue damage, ultimately driving complications such as retinopathy, nephropathy, neuropathy, and cardiovascular disease. Although cellular antioxidant defenses attempt to maintain redox homeostasis, persistent hyperglycemia continuously compromises these protective mechanisms. By systematically evaluating these cellular interactions, this review elucidates the oxidative stress-related pathogenesis of diabetic complications. Ultimately, we revisit the self-perpetuating mechanisms of the hyperglycemia-oxidative stress vicious cycle, offering a structurally unified and highly integrative framework to guide future research.
Objective Dilated cardiomyopathy (DCM) is the most prevalent cardiomyopathy and a leading cause of heart failure worldwide. Midkine (MDK), a conserved heparin-binding growth factor with antiapoptotic, proangiogenic, anti-infective, and proinflammatory functions, has been implicated in inflammatory and fibrotic diseases. Its specific roles and mechanisms in DCM pathogenesis remain unclear. Materials and methods A doxorubicin (DOX)-induced mouse model and DOX-treated HL-1 cardiomyocytes were used to investigate the effects of MDK on DCM progression. Myocardial injury and fibrosis were evaluated using hematoxylin and eosin (HE), Picrosirius Red (PSR), and Masson’s trichrome staining. MDK and Neurexin-1 (NRXN1) localization in cardiac tissues was assessed through immunofluorescence. Cell viability, apoptosis, and surface area were measured by Cell Counting Kit-8 (CCK-8), flow cytometry, TUNEL, and Actin-Red staining. mRNA and protein levels were quantified by RT-qPCR and western blotting. Results MDK expression was significantly elevated in both DOX-induced DCM and HL-1 cells. Exogenous MDK administration aggravated myocardial injury, interstitial fibrosis, and cardiomyocyte apoptosis in vivo, and reduced cell viability while increasing apoptosis and cell surface in vitro. Mechanistically, MDK markedly enhanced DOX-induced upregulation of NRXN1, IL-1β, IL-6, TNF-α, NLRP3, ANP, BNP, NEU1, cleaved caspase-1/caspase-1, cleaved IL-1β, cleaved caspase-3/caspase-3; while attenuating DOX-induced downregulation of NRG1, NEXN, and SYNE1 in vivo and in vitro. Conclusion MDK promotes DCM-associated heart failure through NRXN1 upregulation and enhanced inflammatory responses, highlighting itself as a potential therapeutic target for DCM.
Fibroblast growth factor (FGF) constitutes a diverse family of signaling proteins involved in cell proliferation, differentiation, angiogenesis, lymphangiogenesis and tissue regeneration. Emerging evidence suggests that FGFs may also play a pivotal role in inflammatory bowel diseases (IBD). IBD, comprising Crohn’s disease (CD) and ulcerative colitis (UC), is characterized by chronic, relapsing inflammation of the intestines, manifested primarily by persistent abdominal pain and severe diarrhea. In IBD, by activating specific fibroblast growth factor receptors (FGFRs), FGFs modulate the immune response, promote mucosal healing, and regulate extracellular matrix (ECM) remodeling, thereby maintaining intestinal barrier integrity. Expression levels of several FGFs, including FGF1, FGF2, FGF7, and FGF10 are upregulated in IBD. These alterations often reflect disease exacerbation and correlate with inflammatory markers, suggesting the potential of FGFs as biomarkers. Furthermore, studies in animal models of various gastrointestinal (GI) tract diseases demonstrate promising possibilities for the use of recombinant FGFs as a novel therapeutic approach to managing gut inflammation.
Background Coronary atherosclerosis is a chronic progressive disease driven by lipid metabolism disorders, inflammatory responses, and vascular structural remodeling. Methods This narrative review is based on a literature search of PubMed and Web of Science for articles published up to 2026, using keywords including "coronary atherosclerosis," "stent biomaterial," "vascular remodeling," and "cardiac rehabilitation." Results This manuscript reviews the pathological evolution of the disease at the tissue and cellular level, covering endothelial dysfunction, extracellular matrix (ECM) remodeling, inflammatory cell infiltration, foam cell formation, and plaque calcification. This manuscript then focuses on the tissue dynamic response at the stent-vessel interface after implantation, analyzing acute injury, neointimal hyperplasia, and in-stent neoatherosclerosis (NA) and reviews bioengineering strategies for pro-healing stents. Furthermore, the role of cardiac rehabilitation (CR) in inducing vascular remodeling is explored, emphasizing the regulation of endothelial structure and function by blood flow shear stress, optimization of plaque components, and myocardial repair mechanisms. Conclusions By integrating pathological mechanisms, material interfaces, and rehabilitation interventions, this paper proposes a paradigm shift from "injury response" to "repair promotion." It looks forward to the application of tissue engineering, organoid models, and multi-omics technologies in precision intervention, providing a theoretical basis for personalized treatment and regenerative medicine strategies for coronary heart disease.
