The emergence of carbapenem-resistant Klebsiella pneumoniae highlights the need to evaluate candidate compounds in terms of both KPC-2 recognition and behavior at the Gram-negative outer membrane barrier. Here, the clinically used cyclic boronate inhibitor vaborbactam and two newly synthesized piperazine-hydrazone derivatives, M1 and M2, were compared using molecular docking, replicate atomistic molecular dynamics simulations, MM/PBSA analysis, umbrella-sampling-derived potential of mean force profiles and preliminary phenotypic antibacterial screening. Docking suggested that M2 adopted an initial KPC-2 binding orientation more similar to that of vaborbactam than did M1, whereas replicate MD simulations revealed substantial trajectory-dependent variability and did not support a definitive stability ranking. MM/PBSA analysis showed a more favorable mean enthalpy-like contribution for M2 than for M1; the mean value for M2 approached that obtained for vaborbactam but exhibited greater energetic heterogeneity. In the porin-free membrane model, vaborbactam, M1 and M2 showed ligand-specific inward PMF increases of approximately 14.2, 33.4 and 52.25 kcal/mol, respectively, although these independently referenced profiles should not be interpreted as strictly endpoint-matched permeation barriers. M1 retained substantial hydration and predominantly sampled Lipid A/core contacts, whereas M2 showed pronounced reorientation and a larger inner-leaflet contact population during the externally driven pathway. In preliminary antibacterial screening against K. pneumoniae ATCC 13883, M2 produced a marginally larger inhibition zone than did M1, while both compounds showed identical MIC endpoints. Because KPC-2 production was not established and direct enzyme-inhibition or meropenem-combination assays were not performed, the experimental data do not demonstrate KPC-2 inhibition or superior whole-cell activity. M2 is therefore prioritized for mechanistic follow-up based on its comparatively favorable computational KPC-2 interaction profile rather than on its status as a validated antibacterial candidate.
Perfluorooctanoic acid (PFOA) has been associated with damage to multiple organs, and its effects on the gastrointestinal system have attracted increasing attention. However, the mechanism underlying PFOA-induced intestinal toxicity remains unclear. In this study, we found that PFOA disrupted the intestinal barrier and induced intestinal inflammation in mice, accompanied by increased GzmA expression. Mechanistically, GzmA upregulation promoted intestinal inflammation through the GzmA-PAR1 signaling axis, which enhanced the interaction between caveolin-1 and β-catenin and facilitated the nuclear accumulation of β-catenin. Nuclear β-catenin subsequently upregulated TBX3, which bound to an upstream regulatory of IL24 and promoted IL24 expression following PFOA exposure. To validate the role of GzmA, we knocked down GzmA in normal colonic epithelial cells (NCM460). The results confirmed that GzmA upregulated IL24 expression through the PAR1/β-catenin/TBX3 pathway following PFOA exposure. These findings provide mechanistic insight into PFOA-induced colonic inflammation and highlight the potential utility of this toxicant-based model for investigating molecular targets relevant to intestinal inflammatory diseases.
Acute alcohol exposure is known to impair female reproductive function; however, it remains unclear whether the biological effects of circulating metabolites generated after alcohol ingestion on follicular development differ from those of direct ethanol exposure. Therefore, we compared the effects of serum from mice with acute alcohol exposure and ethanol-supplemented medium on the in vitro development of mouse preantral follicles and explored the underlying mechanisms of these effects. Female Kunming (KM) mice were administered 56% (volume/volume, v/v) ethanol intragastrically to establish an acute alcohol exposure model, followed by serum collection. Preantral follicles were cultured in three systems: foetal bovine serum (FBS) control, serum from alcohol-exposed mice, and medium supplemented with ethanol at an equivalent concentration. Follicular morphology, steroid hormone secretion, gene expression, oxidative stress, apoptosis, oocyte maturation, and subsequent embryonic developmental competence were evaluated. Acute alcohol exposure altered serum composition, and this alteration was characterised by increased ethyl linoleate and reduced cholesterol levels. Compared with the FBS control, follicles cultured with serum from mice exposed to alcohol exhibited enlarged antral cavities and cumulus expansion, whichwas accompanied by meiotic delay at the germinal vesicle (GV) and mid-metaphase I (mid-MI) stages. Alcohol-exposed serum increased follicle-stimulating hormone levels and upregulated forkhead box L2 (Foxl2) expression, thereby promoting oestradiol synthesis while suppressing progesterone secretion during follicular development. In contrast, direct ethanol exposure impaired follicular development by decreasing mitochondrial membrane potential and inducing apoptosis. Although oxidative stress marker levels were elevated in serum from alcohol-exposed mice, oocytes maintained normal reactive oxygen species levels through enhanced antioxidant defence, including increased expression of superoxide dismutase 1 (Sod1), glutathione peroxidase 1 (Gpx1), translocase of inner mitochondrial membrane 23 (Tim23) and polyribonucleotide nucleotidyltransferase 1 (Pnpt1) in oocytes. Notably, oocytes derived from follicles cultured with alcohol-exposed serum exhibited improved embryonic developmental competence after in vitro fertilisation. Thus, serum from mice with acute alcohol exposure exerts biological effects distinct from those of direct ethanol exposure during preantral follicle development. Alcohol-derived serum metabolites partially mitigate ethanol-induced reproductive toxicity through endocrine modulation and activation of antioxidant defence pathways, providing new insights into the complex effects of alcohol metabolism on female reproductive function.
Sucralose is a widely consumed non-caloric sweetener, but its immunotoxicological effects remain poorly defined. Integrated in vitro and in vivo methods were utilized to investigate its immunotoxicological profile. In vitro, sucralose suppressed dendritic cell maturation and IL-6 secretion, promoted the differentiation of naïve CD4+ T cells into Th1 cells, and reshaped cytokine profiles in DC-T cell co-cultures. In the food allergy model of ovalbumin (OVA)-induced BALB/c mice, the treated dose chronic dietary intake of sucralose at 10× the acceptable daily intake (ADI) exerted a surprising immunomodulatory effect, but did not at the dose of ADI. Compared to the OVA allergy group, treatment with the 10×ADI dose of sucralose alleviated allergic symptoms, suppressed serum levels of IgE/IgG1/mMCP-1, restored intestinal barrier function, shifted the Th1/Th2 balance toward Th1 dominance, and inhibited mast cell activation. Multi-omics integration revealed that the 10×ADI dose concurrently modulated immune and microbial axes, with downregulation of pro-allergic mediators (Edn1, Chrm2) within the cAMP signaling pathway and enrichment of beneficial gut microbiota (Lactobacillus), providing mechanistic evidence for a dose threshold at which sucralose engages the gut-immune axis. These findings demonstrated that sucralose exhibited dose-related immunomodulatory activity, providing a more nuanced basis for some food additives safety assessment.
The arginine-glycine-aspartic acid (RGD) tripeptide sequence is widely distributed in extracellular matrix proteins enabling efficient targeting of integrin receptors that are overexpressed on the surface of tumor cells, thereby facilitating the selective delivery of therapeutic molecules to tumor tissues. Cyclic RGD peptides are widely recognized for their higher target affinity relative to linear counterparts, and numerous RGD-based peptides have been evaluated in clinical trials. In this study, we report the synthesis and bioactivity of a novel cyclic RGD peptide incorporating a conformationally constrained isoindolinone scaffold. The target cyclopeptide was prepared via an intramolecular photoinduced single-electron transfer cyclization reaction, and the absolute configuration was unambiguously determined through combined experimental and theoretical electronic circular dichroism (ECD) analyses. We also systematically screened potential targets of the prepared peptide using pharmacophore mapping (TargetNet) combined with reverse molecular docking (GalaxySagittarius-AF), which predicted high binding affinity of the synthesized cyclic peptide for the integrin αvβ6 receptor. This finding was further validated through experimental bio-layer interferometry assays, and molecular docking and molecular dynamics simulations. Comprehensive in vitro and in vivo biological evaluations revealed that cyclic peptide effectively suppresses tumor cell proliferation and induces both apoptosis and autophagy.
Plicamycin (PLI) is a small-molecule aureolic acid antibiotic that has been approved for cancer treatment since the 1970s. However, its interaction with cardiovascular biological systems during myocardial ischemia-reperfusion injury (MIRI) remains largely unknown. This study examines its cardioprotective effects and the mechanisms associated with cAMP response element-binding protein (CREB) and brain-derived neurotrophic factor (BDNF) in a murine MIRI model. PLI, administered intraperitoneally to mice at 150 μg/kg/day for seven days before cardiac ischemia-reperfusion, attenuated the rise in myocardial infarct size and serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels. PLI also improved cardiac function in MIRI mice, as reflected by higher ejection fraction (EF) and fractional shortening (FS) values. Biochemical assays and dihydroethidium (DHE) staining analyses showed that PLI administration prevented oxidative stress imbalance within the ischemic myocardium following reperfusion. PLI also prevented cardiomyocyte apoptosis within ischemic myocardial tissues by downregulating caspase-3, caspase-9, and Bax and upregulating Bcl-2 expression in MIRI mice. Further investigation showed that I/R injury reduced the expression levels of phospho-CREB and BDNF within the ischemic myocardium, both of which were prevented by PLI. More importantly, pharmacological suppression of CREB by 666-15 (10 mg/kg/day) or BDNF signaling by K252a (25 μg/kg/day) diminished the cardioprotective effects of PLI. Overall, our data suggest that PLI prevents I/R-induced cardiac injury by activating CREB-BDNF signaling, accompanied by suppression of nitro-oxidative stress.
Arsenic exposure increases the risk of bladder cancer. The proteomic analysis of arsenic treated bladder epithelial cells revealed significantly upregulated NMES1 expression, which was further validated in the bladder epithelium of arsenite-exposed mice and SV-HUC-1 cells treated with 0.5 μM arsenite. NMES1 is a nuclear-encoded accessory protein of mitochondrial respiratory chain complex IV. A direct interaction between ALKBH7 and NMES1 was confirmed by both molecular docking and protein interaction assays. ALKBH7, a mitochondrial RNA demethylase, was significantly downregulated in arsenic-exposed bladder epithelial cells. Arsenic-mediated ALKBH7 inhibition elevated the m22G methylation level of tRNAs flanking in the mitochondrial respiratory chain, accompanied by decreased mRNA levels of lysine, arginine, and glycine, as well as reduced protein expression of mt-CO2 and mt-ND3, ultimately causing mitochondrial respiratory dysfunction. In addition, arsenic exposure also enhanced cellular glycolysis, which provides energy support for the proliferation and migration of bladder epithelial cells. siNMES1 alleviated arsenic-induced ALKBH7 downregulation and cell malignant phenotype. Overexpressed ALKBH7 effectively attenuated arsenic-enhanced glycolysis and malignant phenotype. In conclusion, arsenic upregulates NMES1 to suppress ALKBH7 expression that increased mt-tRNA m22G methylation modification and reduces the expression of the mt-CO2 and mt-ND3 subunits, thereby impairing mitochondrial respiratory function. Inhibiting ALKBH7 further promoted glycolytic reprogramming to sustain malignant phenotypes in arsenic treated bladder epithelial cells.
Carbonyl reductase 1 (CBR1) is a cytosolic monomeric enzyme belonging to the short-chain dehydrogenase/reductase superfamily and is ubiquitously expressed in human tissues. It catalyzes the NADPH-dependent reduction of a wide range of carbonyl compounds and plays a major role not only in phase I metabolism of xenobiotics, including drugs, but also in the metabolism of endogenous substrates such as prostaglandin E2, S-nitrosoglutathione, ketosteroids, and lipid peroxidation-derived aldehydes. Notably, glutathione binds near the active site of CBR1 and enhances its reductase activity toward non-glutathionylated substrates. Accumulating evidence indicates that CBR1 is involved in cancer, functioning either as a tumor suppressor or a tumor promoter depending on the cancer type. This dual role suggests that CBR1 functions as a context-dependent regulator of cellular redox adaptation. CBR1 is also implicated in several noncancerous diseases, including neurodegenerative disorders, diabetes, ischemia/reperfusion injury, osteoporosis, ulcerative colitis, and COPD. In this review, we summarize current knowledge on the structure, enzymatic functions, gene regulation, modulators, and polymorphisms of CBR1, and discuss its diverse roles in cancers and noncancerous disorders.
Vanadium compounds are among the most extensively investigated insulin-mimetic agents for diabetes mellitus (DM), with reported effects on glucose homeostasis, lipid metabolism, and insulin-related signaling. However, preclinical evidence remains highly heterogeneous due to differences in compound chemistry and experimental design. Then, this study mapped and synthesized preclinical evidence on the effects of vanadium-based compounds on hyperglycemia and dyslipidemia in animal models of DM. This scoping review followed Joanna Briggs Institute methodology and PRISMA-ScR guidelines. PubMed, Scopus, and Web of Science were searched without date restrictions. Eligible studies included diabetic animal models treated with vanadium compounds reporting glycemic and/or lipid outcomes. Data extraction covered compound characteristics, experimental models, intervention protocols, metabolic outcomes, safety, mechanistic findings, and methodological quality using the SYRCLE risk-of-bias tool. A total of 104 studies published between 1989 and 2024 were included. Streptozotocin-induced diabetes was the predominant model (77/104), and rats, particularly Wistar rats (50/104), were the most frequently used species. Inorganic salts and organic complexes were equally represented (52 each). Oral administration and repeated-treatment protocols predominated (100/104). Most studies reported reductions in fasting glucose, improved glucose tolerance, and favorable modulation of insulin signaling, antioxidant defenses, and hepatic metabolism. Lipid outcomes, assessed in 61 studies, generally demonstrated reductions in triglycerides, total cholesterol, and hepatic lipid accumulation. Sankey analysis identified dominant experimental pathways centered on streptozotocin models receiving oral vanadium treatment, whereas SYRCLE assessment revealed frequent deficiencies in reporting randomization, allocation concealment, and blinding, resulting in predominantly unclear risk-of-bias judgments. Vanadium compounds consistently improved glycemic control and frequently ameliorated diabetic dyslipidemia. Coordination chemistry and ligand design appear to be major determinants of therapeutic performance, although improving methodological rigor and reporting quality will be essential to strengthen the translational value of future preclinical research.
Cadmium (Cd), a well-recognized neurotoxicant, elicits neuronal death and cognitive impairment. Cd cytotoxicity disrupts endoplasmic reticulum (ER) proteostasis, leading to the accumulation of misfolded and unfolded proteins and subsequent ER stress-mediated apoptosis. Endoplasmic reticulum autophagy (ER-phagy) serves as a crucial quality-control mechanism that resolves excessive ER stress and maintains ER homeostasis. However, the precise roles of ER-phagy, ER stress-mediated apoptosis, and their crosstalk in Cd-induced neurotoxicity remain poorly defined. Here, we demonstrated that Cd exposure robustly induces ER stress and the subsequent apoptotic injury in the mouse hippocampus and HT-22 hippocampal neurons. Pharmacological inhibition of ER stress with 4-phenylbutyric acid (4-PBA) effectively rescued Cd-triggered neuronal damage, reduced cell death, and ameliorated Cd-associated cognitive deficits. Notably, Cd exposure leads to pronounced ER-phagy dysfunction, as evidenced by decreased LC3-II accumulation, elevated calnexin levels, and impaired ER-phagy autophagic flux in vivo and in vitro. Furthermore, the ER-phagy receptor TEX264 was downregulated under Cd stress. Importantly, restoration of ER-phagy via overexpressing TEX264 markedly mitigated Cd-elicited ER stress, thereby blocking the downstream apoptotic cascade. Collectively, our findings identify impaired ER-phagy as a previously unrecognized mechanism underlying Cd neurotoxicity, which synergizes with ER stress-mediated apoptosis to promote hippocampal neuronal injury. These results highlight ER quality control pathways as promising therapeutic targets for the intervention of Cd-induced cognitive and neuronal damage.
Bosutinib is a second-generation tyrosine kinase inhibitor (TKI) primarily used for the treatment of chronic myeloid leukemia (CML). The aim of this study was to develop and validate a rapid and accurate ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of bosutinib and its major metabolite M5. Chromatographic separation was performed on an Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) using a gradient elution with acetonitrile and 0.1% formic acid as the mobile phase, and gefitinib was used as the internal standard (IS). Biological analysis showed that both bosutinib (2-400 ng/mL) and its metabolite M5 (1-60 ng/mL) exhibited good linearity in their respective concentration ranges, with correlation coefficients (r2) of 0.996 and 0.993, respectively. Both of them exhibited intra- and inter-day precision (RSD%) below 15% and accuracy (RE%) within ± 15%. Recoveries were all within the range of 85.3-98.0%, and matrix effects were all within the range of 91.6-101.6%. Ultimately, human liver microsomes (HLM) was used to study in vitro metabolic stability, while in vivo pharmacokinetics were evaluated in rats. In vivo pharmacokinetic studies confirmed that this method could be successfully used for the analysis of bosutinib after oral administration of 50 mg/kg in rats. In vitro results showed that bosutinib had the intrinsic clearance (CLint) value of 0.03 mL/min/mg and the half-life (t1/2) value of 79.94 min. This study provides a potential reference for the clinical investigation of bosutinib.
Extracellular vesicles (EVs) serve as key mediators of intercellular communication within the nervous system. An adequate glucose supply is essential for normal brain function; however, cerebral glucose metabolism is progressively impaired in Parkinson's disease (PD). Here, we show that EVs released by microglia exposed to 40 μM paraquat (PQ) are internalized by dopaminergic neurons, where they impair glucose metabolism and induce neurodegenerative injury through the intercellular transfer of tenascin-C (TNC). TNC, which has been proposed as a potential indicator of disease severity, is upregulated following neurological injury and has been implicated in neuronal apoptosis and neuroinflammation. EVs released by microglia exposed to 40 μM PQ reduced neuronal glucose uptake, disrupted glucose metabolism, and exacerbated mitochondrial dysfunction. Proteomic analysis revealed that EVs derived from PQ-exposed microglia were enriched in TNC. Co-immunoprecipitation analysis further demonstrated an interaction between EV-transferred TNC and protein kinase B (AKT) in recipient neurons. Both depletion of TNC from EVs and pharmacological activation of AKT restored glucose uptake in recipient neurons. Collectively, these findings demonstrate that, under PQ exposure, activated microglia impair neuronal glucose metabolism by releasing TNC-enriched EVs. The EV-mediated TNC-AKT/glucose transporter 1 (GLUT1) axis may therefore contribute to PQ-induced dopaminergic neurotoxicity and represent a potential therapeutic target for PD.
Artificial sweeteners, notably aspartame, are common food additives and merging environmental contaminants with potential cardiovascular risks. This study integrated network toxicology and experimental validation to explore aspartame's mechanisms in promoting cardiovascular disease (CVD). Initial toxicity prediction via ProTox 3.0 indicated cardiotoxicity among other toxicities. Potential targets of aspartame were identified using SwissTargetPrediction, ChEMBL, and SEA, while CVD associated targets were retrieved from GeneCards and OMIM and atherosclerosis related targets were obtained from GSE100927 dataset. Core targets were prioritized via STRING and Cytoscape. Functional enrichment analysis using Metascape highlighted key pathways. Molecular docking and dynamics simulations assessed binding affinity and stability. Microarray analysis, followed by in vivo and in vitro studies, validated findings. Results showed that aspartame-induced cardiovascular toxicity involves targeting IL1B, TNF, MMP9, CTSS, and CCR5, and activating cell adhesion molecules, NF-κB, and NOD-like receptor signaling pathways. In mice, aspartame reduced immune-related blood cells, impaired endothelium-dependent vasodilation, and increased vascular adhesion molecule expression. In HUVEC, the same pathways were activated. These findings suggest that aspartame promotes cardiovascular toxicity by targeting specific proteins and modulating inflammatory responses, cell migration, and adhesion via NF-κB and NOD-like receptor pathways. Collectively, this study provides mechanistic insights into aspartame-associated cardiovascular risk, providing a comprehensive mechanistic framework for a more accurate assessment.
Intestinal aging is characterized by impaired intestinal stem cell (ISC) function, reduced mucosal regenerative capacity, and progressive epithelial barrier deterioration. However, the upstream metabolic and epigenetic mechanisms that regulate ISC homeostasis during aging remain poorly understood. This study aimed to determine how SIRT6 regulates ISC homeostasis during intestinal aging and to investigate whether Atractylenolide II (AT-II) can alleviate age-related ISC dysfunction. Jejunal tissues from young and aged mice, intestinal epithelial-specific Sirt6-deficient mice, and 3D intestinal organoids were used to evaluate crypt-villus morphology, ISC activity, and lineage differentiation. Mechanistic analyses included western blotting, immunofluorescence, and retinoic acid (RA) quantification, complemented by pharmacological and rescue experiments targeting RXRα activity and RA metabolic balance. The results showed that SIRT6 protein expression was markedly reduced in the aged jejunum, correlating with a decreased villus-to-crypt (V/C) ratio, impaired ISC proliferation, and altered epithelial differentiation. Intestinal epithelial deletion of Sirt6 recapitulated aging-related intestinal defects, including crypt atrophy and ISC-associated dysfunction. Consistently, aged organoids displayed reduced SIRT6 protein expression, while Sirt6Δ/Δ organoids showed decreased SOX9 and Lgr5 expression. Mechanistically, SIRT6 physically associated with RXRα, and SIRT6 loss or inhibition was associated with increased RXRα acetylation, elevated RXRα abundance, and RA metabolic remodeling. These changes were accompanied by disrupted RA balance and aging-like ISC dysfunction. AT-II partially restored SIRT6/RXRα/RA-related molecular alterations and alleviated ISC-associated dysfunction. This study suggests that the SIRT6/RXRα/RA axis contributes to ISC homeostasis during intestinal aging and may be modulated by AT-II, offering a potential strategy for age-related intestinal barrier damage.
BACKGROUND:Bisphenol A (BPA) is a common environmental endocrine disruptor linked to type 2 diabetes mellitus (T2DM) and colorectal cancer (CRC), but the exact mechanism connecting BPA exposure to their comorbidity is unclear. METHODS:This study integrated network toxicology, single-cell transcriptomic data, molecular simulation, in vitro cell experiments, and patient-derived explant (PDE) models to systematically explore the potential molecular basis underlying the correlation between BPA exposure and T2DM-CRC comorbidity. RESULTS:CXCL8 was identified as the sole overlapping gene shared among the 30 BPA-T2DM-CRC common targets, the PPI-derived hub genes, and the DEGs identified in GSE115313. It was upregulated in T2DM-CRC tissues, associated with obesity and T2DM, and showed strong diagnostic performance in CRC (AUC=0.895). Immune analysis indicated an increased proportion of Tregs in CRC samples from patients with T2DM, while CXCL8 expression correlated positively with M1 macrophage and activated mast cell infiltration. Single-cell transcriptomic data analysis of GSE188711 showed that CXCL8 expression varied along the inferred pseudotime trajectory and appeared enriched in the annotated B-cell population. Molecular simulations showed moderate binding between BPA and CXCL8 (binding energy: -5.7 kcal/mol). BPA treatment boosted CRC cell proliferation, migration, and invasion, while CXCL8 knockdown reduced these activities. The malignant effects of BPA were significantly reduced when CXCL8 was knocked down. Mechanistically, TEAD4 was identified as a potential upstream transcriptional regulator of CXCL8, while increased CXCL8 expression was accompanied by enhanced p38 MAPK pathway activation. CONCLUSIONS:BPA-induced CXCL8 upregulation is associated with CRC malignant progression and enhanced p38 MAPK signaling. TEAD4 may function as an upstream transcriptional regulator of CXCL8, although its direct regulation by BPA remains unclear. Given the clinical correlation between CXCL8 upregulation and T2DM status, this pathway may represent a potential mechanistic link contributing to the frequently observed T2DM-CRC comorbidity.
27-Hydroxycholesterol (27-HC), as a major cholesterol metabolite, has been shown to be associated with tumor biology, but its role in ovarian cancer remains unclear. This study integrates network toxicology, multi-cohort transcriptomic validation, and single-cell RNA sequencing to systematically investigate the potential role of 27-HC in ovarian cancer and identify key molecular mediators. Among 51 overlapping targets, MMP9 was identified as a robust candidate and consistently validated in independent datasets. Single-cell analysis revealed that MMP9 exhibited distinct expression patterns across different cell populations and was associated with extracellular matrix-related processes. Molecular docking and dynamic simulations confirmed a stable interaction between 27-HC and MMP9. Functionally, 27-HC significantly promoted ovarian cancer cell proliferation, cell cycle progression, and malignant phenotypes in vitro, and facilitated tumor growth in vivo. Concurrently, 27-HC treatment significantly upregulated MMP9 expression at both the mRNA and protein levels in ovarian cancer cells. In summary, these findings provide evidence supporting a potential role of 27-HC in ovarian cancer and its association with MMP9 upregulation, highlighting MMP9 as a potential mediator linking cholesterol metabolism to tumor progression.
Vitamin supplements are frequently utilized as adjunctive therapies in epilepsy care, yet the potential for toxic drug-nutrient interactions remains poorly defined. Using a pentylenetetrazol (PTZ) zebrafish seizure model, we conducted an exploratory screening of a multivitamin formulation containing vitamins B6, B9, D3, and E, together with CoQ10, in combination with common antiseizure medications (ASMs). This screening identified a severe, unexpected lethal interaction between pyridoxine (PN) and valproic acid (VPA). Co-exposure resulted in rapid, dose- and time-dependent mortality in both larval and adult zebrafish. Mechanistic investigations indicated that the calcium-dependent apoptosis, rather than necroptosis, contributes substantially to this toxicity. In vivo calcium imaging revealed marked cytosolic Ca2+ accumulation, which was alleviated by calcium chelation and by pharmacological inhibition of endoplasmic reticulum Ca2+ release through inositol 1,4,5-trisphosphate receptors and ryanodine receptors. PN/VPA co-exposure also caused pronounced cardiac dysfunction, characterized by reduced heart rate, ejection fraction, and cardiac output, together with apoptotic signals in cardiac tissue. Notably, no comparable cytotoxicity was observed in cultured zebrafish cardiomyocytes or mammalian cell lines, suggesting that organism-level or tissue-integrated factors may contribute to the observed toxicity. Supplemental pyridoxal 5'-phosphate (PLP) effectively rescued PN/VPA-induced mortality, supporting the possible involvement of altered vitamin B6/PLP-related cellular processes. Together, these results identify a previously unrecognized PN/VPA hazard and highlight the zebrafish as a sensitive in vivo platform for detecting complex drug-nutrient toxicities.
Benzo[a]pyrene (BaP), a representative polycyclic aromatic hydrocarbon, is a pervasive environmental pollutant with emerging cardiovascular toxicity. However, the molecular mechanisms linking BaP exposure to pathological cardiac hypertrophy remain incompletely understood. Here, we integrated target-prioritization analyses with in vivo and in vitro validation to investigate whether growth arrest-specific 6 (GAS6) mediates BaP-induced cardiac remodeling. Target integration and Mendelian randomization analysis nominated GAS6 as a biologically plausible mediator associated with cardiac hypertrophy susceptibility. Structural modeling and molecular dynamics simulations further suggested a stable and structurally feasible interaction between BaP and GAS6. In a chronic BaP-exposure mouse model, BaP induced myocardial hypertrophy, myocardial fiber disorganization, and interstitial remodeling, accompanied by increased expression of GAS6, HIF-1α, VEGF-A, GLUT1, and LDHA in cardiac tissues. AAV9-mediated GAS6 silencing partially alleviated these histopathological abnormalities and reduced hypoxia- and glycolysis-associated protein expression. Consistently, in H9C2 and AC16 cardiomyocytes, BaP upregulated hypertrophic markers, including NPPA, NPPB, and MYH7, and activated mTOR/HIF-1α-associated glycolysis-related metabolic signaling. GAS6 knockdown attenuated BaP-induced hypertrophic gene expression and reduced the expression of mTOR/HIF-1α-associated glycolysis-related proteins. Collectively, these findings indicate that GAS6 contributes to BaP-induced pathological cardiac hypertrophy by linking environmental toxicant exposure to mTOR/HIF-1α-associated glycolytic remodeling. This study provides mechanistic insight into BaP-associated cardiovascular toxicity and identifies the GAS6/mTOR/HIF-1α axis as a potential mechanistic target in environmentally induced cardiac remodeling.
