Insomnia is closely associated with immune dysregulation, yet the overall pattern of peripheral-central immune disequilibrium and its underlying molecular basis remains incompletely understood. To characterize the peripheral-central immune features associated with insomnia, identify key immune cell populations and core molecular programs, and prioritize candidate therapeutic compounds with preliminary experimental validation. Peripheral blood bulk transcriptomic dataset GSE208668 was analyzed using differential expression analysis, weighted gene co-expression network analysis (WGCNA), and functional enrichment analysis to identify insomnia-associated genes. Protein-protein interaction network analysis and machine learning models were then applied in independent peripheral blood datasets to refine core genes. Immune deconvolution and peripheral blood single-cell transcriptomic dataset GSE213496 were used to determine the immune-cell context, cell-type localization, and intercellular communication features of these genes. The brain single-cell transcriptomic dataset GSE137665 was further analyzed to assess central alterations. Drug prediction, molecular docking, and molecular dynamics simulations were performed to prioritize candidate compounds, followed by in vitro validation of resveratrol in an LPS-induced THP-1 macrophage model. A total of 5,321 differentially expressed genes associated with insomnia were identified, and weighted gene co-expression network analysis highlighted the turquoise and blue modules as key insomnia-related modules. Integrative analysis yielded 390 intersecting genes enriched mainly in immune, inflammatory, and oxidative stress-related pathways. Protein interaction analysis and machine learning further identified six refined core genes: FN1, HMOX1, HSP90AA1, IL10, MYD88, and NFE2L2. Because IL10 was not stably detected in the single-cell datasets, the remaining five genes were used for downstream single-cell analyses. Immune deconvolution suggested selective peripheral immune remodeling in insomnia, characterized by increased resting CD4 memory T cells and M2 macrophages, together with reduced activated NK cells. Peripheral single-cell analysis showed that HMOX1, HSP90AA1, MYD88, and NFE2L2 were mainly enriched in neutrophils, inflammatory macrophages, conventional dendritic cells, and selected lymphocyte populations, whereas FN1 showed a more restricted distribution pattern. CellChat analysis indicated enhanced intercellular communication under the sleep deprivation-related condition. In contrast, brain single-cell analysis revealed comparatively modest but detectable central alterations, including enrichment of ependymal cells, slight increases in excitatory neurons, brain endothelial cells, and choroid plexus stromal fibroblasts, together with heterogeneous expression of the core hub genes and selective rewiring of intracerebral communication networks. Drug prediction consistently prioritized quercetin and resveratrol, and structural analyses supported stable interactions with key targets. In THP-1 macrophages, resveratrol downregulated MYD88 and HSP90AA1 while further upregulating HMOX1. Insomnia appears to be associated predominantly with a peripheral-centered immune disequilibrium pattern, characterized by selective remodeling of innate immune-related populations, enhanced inflammatory and oxidative stress programs, and increased intercellular communication. The MYD88-HMOX1-HSP90AA1-NFE2L2 axis may represent a key molecular program linking inflammatory activation and oxidative stress adaptation. Resveratrol was identified as a potential compound and has been validated through preliminary in vitro experiments.
Gouty arthritis (GA) is an inflammatory joint disease caused by the deposition of monosodium urate (MSU) crystals within the joint space and surrounding tissues. In traditional Chinese medicine, Rhizoma Drynariae (Gusuibu) has long been widely used in the clinical treatment of GA, and flavonoids are considered its key bioactive constituents. This research employed network pharmacology to construct a component-target network of total flavonoids of Rhizoma Drynariae (TFRD) against GA, thereby identifying key components, core targets, and related pathways. Rat models were established by intra-articular injection of a monosodium urate crystal suspension and treated with TFRD or the positive control, colchicine, by oral gavage. After sample collection, network pharmacology-based prediction results were subsequently validated using rat serum metabolomics, enzyme-linked immunosorbent assay (ELISA), and Western blot analysis. Network pharmacology analysis indicated that the anti-GA effects of TFRD are mediated through key targets, including IL6, AKT1, TNF, EGFR, JUN, and PTGS2, and are mainly associated with inflammation, immune, and apoptosis-related pathways, such as the IL-17, TNF, NF-κB, MAPK, PI3K-AKT, JAK-STAT, and T-cell receptor signaling pathways. Similarly, metabolomics also uncovered the pivotal roles of the inflammatory response. Hematoxylin and eosin (H&E) staining confirmed that TFRD reduced infiltration of inflammatory cells. ELISA assay confirmed that the TFRD group significantly inhibited the expression of inflammatory factors TNF-α, IL-6, and IL-17A in synovial tissue. Western blot analysis revealed that TFRD inhibited the GA-induced hyperphosphorylation of AKT, MAPK p38, and NF-κB p65 in rat synovial tissue. TFRD can effectively ameliorate the inflammation-triggered changes in the GA rats by directly modulating related inflammatory factors and pathways.
Acute kidney injury (AKI) is a common and serious complication after cardiac arrest, affecting more than 30% of initially successfully resuscitated patients, with around 10% receiving Kidney Replacement Therapy (KRT). Currently, treatment options to prevent renal complications are limited. In this predefined sub-study of the Sedation, TEmperature and Pressure after Cardiac Arrest and REsuscitation (STEPCARE) trial, we evaluate the effects of sedation depth, fever management strategies, and mean arterial pressure (MAP) targets on the risk of major adverse kidney events (MAKE) in post-cardiac arrest patients. STEPCARE-MAKE is a predefined prospective sub-study of the international, randomized clinical STEPCARE trial. The main trial will enroll 3500 patients and employs a 2 × 2 × 2 factorial design, randomizing participants across three interventions: (1) deep sedation for 36 h or minimal sedation, (2) fever management with or without a feedback-controlled device, and (3) MAP target ≥ 65 mmHg or ≥ 85 mmHg. The primary outcome is the composite outcome MAKE, which includes death from any cause by day 30, initiation of KRT, or persistent renal dysfunction, defined as the final creatinine value ≥ 200% of the baseline at the time of discharge from the primary hospital or at day 30, whichever occurs first. Differences between baseline and the highest in-hospital creatinine, baseline and the last measured in-hospital creatinine, baseline and 72-h creatinine, and baseline and the highest creatinine within 72 h will be reported as secondary outcomes. The findings of this sub-study will provide new evidence on the renal effects of the STEPCARE trial interventions and may inform the future of individualized kidney-protective treatment approaches.
Gastric cancer (GC) is one of the main causes of cancer-related global mortality. The emergence of drug resistance and toxicity in current therapies highlights the need for novel treatment strategies. Quercetin, a natural flavonoid, has demonstrated anticancer activity; however, its molecular mechanism, particularly its effect on key targets in GC, remains underexplored. A comprehensive in silico and in vitro methods were used to elucidate the anticancer potential of Quercetin. Network pharmacology analysis was used to identify potential GC-related targets, followed by molecular docking and 200 ns molecular dynamics (MD) simulations to evaluate the binding affinity and stability of the Quercetin-target complex. In vitro experiments, including gene expression analysis and fluorescence binding assays, were conducted using AGS gastric cancer cells to validate the computational findings.nsulin-like growth factor 1 (IGF1) emerged as a key hub gene associated with GC progression. Molecular docking predicted a favorable interaction between Quercetin and IGF1, with a docking score of - 6.3 kcal/mol and multiple hydrogen-bond interactions. MD simulations confirmed the stability of the Quercetin-IGF1 complex, with reduced RMSD values (0.48 nm vs. 0.63 nm for unbound IGF1), favorable free energy profiles, and stable hydrogen bonding. In vitro studies demonstrated a significant downregulation of IGF1 mRNA expression (p < 0.001) and a dose-dependent inhibition of IGF1 activity by Quercetin. The integration of network pharmacology, computational modeling, and experimental validation suggests that Quercetin may modulate the IGF1 signaling axis and influence IGF1-associated pathways in gastric cancer. The stable binding and significant inhibitory effect observed suggests that Quercetin may interrupt IGF1-mediated signaling pathways involved in tumor growth and survival. This study identifies Quercetin as a potential modulator of IGF1-associated signaling pathways with significant therapeutic promise for gastric cancer. The findings provide mechanistic insights supporting the further development of Quercetin as a targeted therapy for IGF1-driven malignancies.
Diabetic nephropathy (DN) is a major microvascular complication of diabetes mellitus and the leading cause of end-stage renal disease. Oxidative stress and inflammation are central drivers of DN progression, yet no effective therapies exist to prevent or delay renal injury. This study investigated the renoprotective effects of glycine (GLY), N-acetylcysteine (NAC), and their combination administered at early versus late stages of streptozotocin induced diabetes. Forty-eight male Wistar rats (n = 48) were allocated into five groups: healthy controls (Group 1, n = 6), untreated diabetic rats (Group 2, n = 6), and three treatment groups (Groups 3-5, each n = 12). Diabetes was induced by a single intraperitoneal streptozotocin injection (55 mg/kg). Group 3 received NAC (100 mg/kg), Group 4 received GLY (250 mg/kg), and Group 5 received NAC + GLY. Each treatment group was subdivided into early (6 week, n = 6) and late (12 week, n = 6) intervention subgroups. Treatments were administered orally. Renal tissue was evaluated using classic histology, geometric morphometric analysis, and biochemical assays of superoxide dismutase (SOD) and myeloperoxidase (MPO). Statistical analyzes were performed using ANOVA with appropriate post hoc tests (p < 0.05). Untreated diabetic rats (Group 2) showed significantly decreased SOD activity, increased MPO levels, marked mesangial matrix expansion, glomerular hypercellularity, tubular epithelial degeneration, and interstitial inflammation with fibrosis. NAC (Group 3) and GLY (Group 4) each improved oxidative stress markers and partially restored glomerular and tubular morphology, with early treatment subgroups exhibiting more substantial benefit than late subgroups. The combined NAC + GLY therapy (Group 5) demonstrated the strongest renoprotective effect, preserving renal structure and biochemical parameters closest to healthy controls. To conclude, early combined administration of glycine and N-acetylcysteine yields superior protection against diabetes-induced renal injury compared with individual treatments. These findings support the therapeutic potential of antioxidant-amino acid combinations in preventing or delaying diabetic nephropathy.
Aberrant epidermal growth factor receptor (EGFR) signaling drives multiple cancers, but the clinical effectiveness of EGFR inhibitors is limited by relapse, toxicity, and mutation-associated resistance. This study applied an integrated computational drug-repurposing workflow combining machine learning-based potency prediction, structure-based docking, and molecular dynamics simulation to prioritize approved DrugBank compounds for mutant EGFR evaluation. EGFR bioactivity data from ChEMBL were curated, standardized, converted to pIC50 values, and represented using SwissADME physicochemical descriptors and Morgan fingerprints. Among the evaluated regression models, ExtraTrees performed best and was selected for screening. Using ECFP plus descriptor features, ExtraTrees achieved R2 = 0.71 ± 0.02 and RMSE = 0.74 ± 0.02 under random splits, and retained useful performance under Murcko scaffold splits with R2 = 0.55 ± 0.01, RMSE = 0.90 ± 0.02, and Spearman ρ = 0.74 ± 0.02. The model was used to screen approved DrugBank compounds, prioritize 500 candidates, and guide docking against EGFR L858R/T790M/C797S (PDB: 6LUD). Docking identified Abemaciclib (-9.65 kcal/mol), Crizotinib (-8.29 kcal/mol), and Avapritinib (-8.10 kcal/mol) as favorable candidates. Abemaciclib scored slightly more favorably than Osimertinib (-9.44 kcal/mol) in this non-covalent docking setup. Subsequent 100 ns molecular dynamics simulations refined the ranking, with Crizotinib and Avapritinib showing more favorable dynamic profiles, while Abemaciclib showed greater ligand mobility. Oncology drugs were significantly enriched among the top 50 docked hits relative to the scored DrugBank background. These results support ML-guided docking and MD refinement as a practical strategy for prioritizing repurposing candidates for experimental EGFR validation.
Spinal cord regeneration after injury remains a major clinical challenge due to the persistent inflammatory microenvironment associated with immune cell infiltration. Withaferin A (WA), a natural anti-inflammatory steroidal lactone with potential NF-κB-modulating activity, was identified by network-based screening as a candidate for spinal cord injury (SCI) and selected for further study. This study aimed to identify and evaluate potential anti-inflammatory small-molecule therapeutics for SCI using a network-based drug screening strategy. Experimental study combining computational drug screening, in vitro macrophage assays and in vivo SCI mouse models. Network pharmacology identified 296 candidate drugs targeting 113 SCI-related genes. Five top candidates, selected for chemical diversity and accessibility, were tested in LPS-stimulated in vitro models for effects on macrophage polarization and cytokine release. Molecular docking was used to predict drug-target interactions. The lead compound, WA, was then evaluated in SCI mice for inflammation, angiogenesis, neuroregeneration and motor recovery. WA showed the strongest anti-inflammatory activity, dose-dependently inhibiting LPS-induced M1 polarization and TNF-α/IL-6 secretion while promoting M2 polarization and IL-4/IL-10 secretion. Integrated computational and experimental analyses identified Cys160 in a hydrophobic pocket of NF-κB p65 as the covalent binding site of WA. In vivo, WA modulated macrophage polarization, reduced inflammatory mediator secretion, increased VEGF immunoreactivity and promoted neuroregeneration and motor recovery. WA exerts significant anti-inflammatory and neuroregenerative effects in SCI models. It also increases VEGF immunoreactivity, suggesting a potential pro-angiogenic effect after injury. These findings support WA as a promising therapeutic candidate for SCI.
Maternal sleep deprivation (MSD) is a common but usually unnoticed issue during pregnancy, and in recent years, it has been increasingly recognised as an important prenatal stressor that may adversely influence maternal physiology, placental function, and fetal neurodevelopment. Sleep disturbances during pregnancy, including reduced sleep duration, fragmented sleep, poor sleep quality, circadian disruption, and rapid eye movement sleep restriction, have been associated with altered hypothalamic-pituitary-adrenal axis activity, systemic inflammation, oxidative stress, and impaired circadian regulation. Emerging evidence from clinical and preclinical studies suggests that these alterations may affect fetal neurogenesis, synaptic development, neuroimmune signaling, and maturation of brain circuits involved in cognition and emotional regulations. Within the framework of the Developmental Origins of Health and Disease, maternal sleep disturbances may contribute to epigenetic modifications, mitochondrial dysfunction, microglial activation, and altered neuroplasticity-related pathways, which are increasingly implicated in long-term neurological vulnerability. Experimental findings further indicate that prenatal sleep disruption may impair offspring cognitive performance, emotional behavior, and stress responsiveness, while potentially influencing biological pathways associated with brain aging-related processes. However, the extent to which MSD directly contributes to pathological brain aging in humans remains incompletely understood. Factors such as timing and duration of exposure, sex-specific responses, and postnatal environmental conditions may further influence offspring outcomes. Therefore, this narrative review critically summarizes current evidence regarding MSD and examines the molecular, cellular, and neurodevelopmental mechanisms through which prenatal sleep disturbances may influence long-term neurological health and vulnerability to brain aging-associated alterations in offspring.This graphical abstract illustrates the mechanistic framework connecting maternal sleep deprivation to the developmental programming of brain aging in offspring. [ MSD: maternal sleep deprivation; DOHaD: Developmental Origins of Health and Disease; 11β HSD2: 11β hydroxysteroid dehydrogenase type 2; ROS: reactive oxygen species; REM: rapid eye movement; HPA axis: hypothalamic pituitary adrenal axis; BDNF: brain derived neurotrophic factor].
Antibiotic-associated diarrhea (AAD) remains a common complication of antibiotic therapy. While probiotics show therapeutic potential, the novel strain Bacillus subtilis THC1I has not been previously evaluated for AAD treatment. This study aimed to assess the effects of B. subtilis THC1I on intestinal barrier integrity and gut microbiota dysbiosis in an AAD mouse model. Fifty mice were randomized into five groups (n = 10): control, model, positive control (Enterogermina®, B. clausii), and a B. subtilis THC1I spore suspension at 0.82 × 10⁹ and 1.64 × 10⁹ CFU/kg/day. AAD was induced with gentamycin sulfate and cefradine for five consecutive days. Outcomes included clinical symptoms, hematological and biochemical parameters, colonic macroscopic and histopathological indices, inflammatory cytokines, and gut microbiota analyzed by 16S rRNA sequencing. B. subtilis THC1I significantly improved body weight, water intake, fecal scores, and fecal water content. Treatment restored electrolyte balance (sodium and potassium), reduced white blood cell counts, and decreased relative colon weight and inflammation scores. Histopathological analysis revealed restored epithelial architecture and increased goblet cell density. Pro-inflammatory cytokines (TNF-α, IL-1β) were significantly reduced. B. subtilis THC1I partially improved microbial diversity (Shannon index) and modulated microbiota composition at both phylum and genus levels, decreasing pathogenic bacteria (Proteobacteria phylum: Escherichia-Shigella, Klebsiella, Salmonella; Firmicutes phylum: Clostridioides), and modulating the dysbiotic overgrowth of the beneficial commensal Blautia, while promoting beneficial bacteria (Bacteroidota phylum: Bacteroides, Muribaculaceae; Firmicutes phylum: Lachnospiraceae, Lactobacillus). B. subtilis THC1I demonstrates restorative effects on intestinal barrier damage and gut microbiota dysbiosis in AAD mice, supporting its potential as a therapeutic candidate for clinical application.
Those affected by Type 2 diabetes (T2DM) often encounter microvascular harm, and within this context, the kidneys are especially prone to damage. Among the approaches to manage such microvascular complications, Shenqi Tangluo Pill (SQTLW), a traditional Chinese herbal formulation, is commonly utilized in clinical settings for preventing and addressing microvascular issues linked to T2DM. Nonetheless, the detailed impacts and working principles of this herbal formulation in alleviating kidney damage caused by diabetes are still not clearly understood. The aim was to confirm the kidney-protective impacts of SQTLW on T2DM mice, as well as to clarify the fundamental mechanisms related to intestinal microbiota and the microbe-derived metabolite trimethylamine N-oxide (TMAO), especially under the condition of intestinal flora depletion induced by a combination of broad-spectrum antibiotics (ABx). Diabetic db/db mice received sustained SQTLW intervention over an 8-week period. Following the 8-week intervention, evaluations were performed on fasting blood glucose levels (FBG), body mass, kidney function, and histopathological changes in renal tissue. Additionally, the makeup of gut microbial communities and the circulating concentrations of metabolites linked to TMAO were assessed. To assess the integrity of the intestinal barrier, histopathological examination of colonic tissue and expression profiles of zonula occludens protein-1 (ZO-1) and Occludin were performed. Expression levels of NOD-like receptor pyrin domain-containing 3 (NLRP3), Smad family member 2/3 (Smad2/3), fibronectin (FN), and α-smooth muscle actin (α-SMA) in renal tissue were examined using immunodetection methods. Next, db/db mice were given a combination of ABx and SQTLW treatments to further examine if the therapeutic outcome is dependent on the regulation of gut microorganisms. SQTLW notably lowered FBG and body weight in db/db mice, ameliorated renal dysfunction, and alleviated histopathological injury of renal tissue, with a notable dose-response relationship. 16S rRNA gene sequencing-based profiling revealed that SQTLW substantially altered the compositional structure of distinct bacterial genera from the phyla Bacteroidota and Firmicutes, particularly Muribaculaceae and Lachnospiraceae. Colonic histopathology and molecular detection indicated that SQTLW effectively preserved intestinal barrier architecture. Targeted metabolomics via LC-MS/MS demonstrated that SQTLW significantly modulated TMAO-related plasma metabolites, including TMAO, choline, and creatinine. Moreover, SQTLW significantly downregulated renal expression of NLRP3 and Smad2/3, thereby attenuating fibrosis-related injury. Importantly, further experiments confirmed that depletion of gut microbiota by ABx partially attenuated the renoprotective effects of SQTLW in db/db mice. SQTLW confers protective effects against renal injury secondary to T2DM, potentially via modulation of gut microbial composition, preservation of intestinal barrier function, and regulation of TMAO-related metabolic pathways.
The receptors of the tumor necrosis factor (TNF) receptor superfamily (TNFRSF) are of overwhelming scientific and clinical relevance and stand at the center of intensive basic and translational research efforts. TNFRSF receptors (TNFRs) are engaged by membrane-bound ligands of the TNF superfamily (TNFSF) and, in some cases, by soluble ligand molecules released from the membrane-bound TNFSF ligand (TNFL) molecules. The development of recombinant TNFL-based TNFR agonists for research and especially therapeutic purposes is highly "individualized", as ligand type-specific hurdles must be overcome in terms of stability, manufacturability, TNFR-specificity and need for oligomerization. TNFR-specific antibodies can also show agonistic activity, but this agonism typically requires FcγR-binding, resulting in a reciprocal conditional bispecific FcγR/TNFR agonism not useful for the study or exploitation of pure TNFR agonism. Some antibodies trigger intrinsic TNFR agonism independent from FcγR-binding, but the rational development of such antibodies is poorly predictable and furthermore challenging due to isotype- and epitope-requirements and poor specific activity when benchmarked with FcγR-bound anti-TNFR antibodies.Using a series of nanobodies (or single-domain antibodies (sdAbs) or variable heavy domain of heavy chains (VHHs)) specific for the TNFRSF members 41BB, BCMA, CD40, CD95, TRAILR2/DR5, GITR, OX40, TNFR1 and TNFR2, we show here that genetic fusion of single-chain encoded triplets of these nanobodies with oligomerizing protein scaffolds regularly results in potent hexa-, nona- and dodecavalent agonists inducing TNFR signaling with EC50-values in the sub-nanomolar range. The oligovalent nanobody formats described exhibit superior CMC properties and enable the simple generation of highly active TNFR agonists from virtually any TNFR-specific nanobody.
Cisplatin is a key treatment for head and neck squamous cell carcinoma (HNSCC), but the development of resistance severely limits its effectiveness. The molecular determinants underlying cisplatin resistance in HNSCC remain unclear. Multiple databases were used to screen the core genes related to cisplatin resistance in HNSCC patients. Tumor tissue samples from HNSCC patients were collected and the expression of FOXA2 was verified through various pathological tests to establish the correlation between FOXA2 expression and the clinical characteristics of the patients. The in vitro and patient-derived organoids (PDOs) models were used to verify the regulatory effect of FOXA2 on the cisplatin resistance of HNSCC. Transcriptome sequencing combined with multi-omics analysis demonstrated that LAMC2 is a downstream target of FOXA2 in regulating cisplatin resistance. Bioinformatic screening of cisplatin-resistant cohorts revealed that FOXA2 was the only gene significantly associated with poor survival outcomes in TCGA-HNSCC patients. Transcriptomic profiling and pathway enrichment analyses revealed the activation of the PI3K/AKT signaling cascade. We identified LAMC2 as a direct transcriptional target of FOXA2. Chromatin immunoprecipitation and luciferase reporter assays confirmed FOXA2 binding to the LAMC2 promoter, resulting in transcriptional activation. FOXA2-mediated upregulation of LAMC2 increased PI3K and AKT phosphorylation, and LAMC2 overexpression reversed the impaired malignant phenotypes caused by FOXA2 silencing. In xenograft models and PDO systems, FOXA2 overexpression reduced responsiveness to cisplatin, whereas FOXA2 inhibition significantly increased therapeutic sensitivity. Our research has identified a previously unrecognized regulatory axis involving FOXA2, LAMC2, and PI3K/AKT, which plays a crucial role in the progression and resistance to cisplatin in HNSCC. Therefore, targeting the FOXA2-LAMC2 axis may represent a novel therapeutic strategy to overcome cisplatin resistance in HNSCC.
The human digestive system is first colonized at birth with many microorganisms that impact on the overall health of the individual via various metabolic, immune, and neuroendocrine pathways. Such microorganisms are referred to as probiotics, according to the FAO and WHO, and they exert effects on host and microbiota interaction primarily through strain-specific bioactive metabolites that act both locally and systemically. The current review will detail the various types of bioactive metabolites and their roles in regulating redox homeostasis via the reduction of reactive oxygen species (ROS) and increases in antioxidant defenses. These include catalase, glutathione peroxidase, and superoxide dismutase, which are believed to be responsible for reducing inflammation and improving epithelial barrier function. In addition to the traditional single-strain approach to probiotics, synbiotics represent an alternative strategy to improve microbial survival, functional stability, and microbiome homeostasis. Synbiotics consist of a combination of probiotic organisms and selective prebiotics that enhance the survivability and functional capacity of the individual strains and thus the overall functional resilience of the gut microbiome. Emerging evidence from mechanistic, experimental, and clinical studies demonstrates the increasing relevance of synbiotics in both therapeutic and translational contexts in the modern healthcare system.
This study investigated the effects of dietary sodium butyrate supplementation on Nile tilapia, with emphasis on growth performance, biochemical alterations, histological characteristics, and gene expression patterns under both normal conditions and glyphosate exposure. A total of 90 Nile tilapia with an initial mean body weight of 7.93 ± 0.026 g were randomly allocated into two experimental groups, with each group comprising three replicate units. The control group was fed a basal diet, while the second group received 1.5 g/kg of sodium butyrate mixed into their food for 8 weeks. Afterward, each group was further divided into two: one without any challenge and the other exposed to 0.6 mg/L GLY. The findings indicated that fish fed with SB exhibited a significant improvement (p < 0.05) in final body weight (FBW), weight gain percentage (WG %), specific growth rate (SGR), protein efficiency ratio (PER), and survival rate, along with a reduction in feed conversion ratio (FCR) compared to the control group. Additionally, there was an enhancement in hepatic antioxidant capacity, along with a downregulation of hepatic ilgf1bp and myostatin. Fish subjected to GLY displayed the highest activities of ALT and AST, elevated levels of BUN and creatinine, and a decrease in lysozyme activity. Sodium butyrate supplementation mitigated glyphosate-induced hepatic and renal impairment and regulated the mRNA expression of intestinal tight junction-associated and apoptosis-related genes. These findings suggest that dietary sodium butyrate may be used as a promising alternative feed additive in sustainable Nile tilapia aquaculture. Its inclusion could help protect fish against glyphosate-associated stress, improve growth performance, enhance antioxidant and immune-related biochemical responses, and provide anti-inflammatory and anti-apoptotic benefits under both basal conditions and following glyphosate exposure.
Background/Objectives: Excitotoxicity, primarily caused by excessive glutamate signaling, is a significant contributor to the aetiology of several neurological disorders. Docosahexaenoic acid (DHA), a long-chain omega-3 polyunsaturated fatty acid, is known for its neuroprotective properties, including antioxidants and anti-inflammatory effects. However, the existing literature has not sufficiently reviewed its specific role in glutamate-induced excitotoxicity. This systematic review aimed to provide comprehensive information from the literature on the neuroprotective effects of DHA in models of glutamate-induced neurotoxicity. Methods: A systematic search was conducted in the Cochrane Library, Scopus, Web of Science, PubMed, ScienceDirect, and Google Scholar, following PRISMA 2020 guidelines. The following keywords were used: DHA OR docosahexaenoic acid AND excitotoxicity OR glutamate-induced excitotoxicity OR glutamate-induced neurotoxicity. A total of 475 articles were screened, and 13 original articles published between 2003 and 2025 were included for data extraction. These studies included nine in vivo animal studies, three ex vivo studies, and one in vitro study. The risk of bias was assessed using SYRCLE's methodology. Results: Our findings demonstrate that DHA provides substantial neuroprotection against excitotoxicity through antioxidative, anti-inflammatory, and anti-apoptotic mechanisms. Furthermore, DHA enhances neuronal function and cognitive performance by modulating neurotransmitter levels and glutamate-related signaling pathways. Despite these positive outcomes, heterogeneity across studies suggests that the neuroprotective properties of DHA may be affected by various parameters, such as the source of DHA, treatment dose and duration, age and experimental design. Conclusions: Although previous studies have demonstrated the benefits of DHA in preclinical and clinical settings of neurological disorders, further clinical studies focusing on the modulation of excitotoxicity by DHA are needed to validate its translational efficacy and therapeutic significance.
To evaluate whether cumin (Cuminum cyminum) oil attenuates nandrolone decanoate (ND)-induced hepatic alterations in rats. Eighty male Sprague-Dawley rats were randomized into six groups: control, cumin oil alone, ND low dose (10 mg/kg/week), ND high dose (20 mg/kg/week), ND low dose + cumin oil, and ND high dose + cumin oil. Cumin oil was administered orally at 400 mg/kg/day for 4 weeks. Outcomes included relative liver weight index, serum ALT/AST, total bilirubin, lipid profile, and blinded histopathology. Both ND doses increased ALT/AST and bilirubin levels and worsened the lipid profile compared with controls, with more pronounced and significant alterations in the high-dose ND group. Co-administration of cumin oil attenuated ND-associated elevations in liver enzymes and bilirubin, improved lipid parameters, and was associated with reduced histological damage compared with ND alone. Interestingly, cumin oil alone increased ALT/AST and lipid parameters compared with controls, although liver architecture remained unremarkable on H&E. In this rat model, cumin oil co-administration partially attenuated ND-induced hepatic biochemical, lipid, and histological alterations. However, cumin oil alone increased ALT/AST and lipid parameters despite unremarkable H&E morphology. Therefore, the present findings should be interpreted as evidence of context-dependent attenuation during ND exposure, not as proof of an independent hepatoprotective or lipid-lowering effect of cumin oil in healthy rats. Dose-ranging, safety evaluation, batch-specific chemical profiling, preparation standardization, and mechanistic studies incorporating antioxidant/oxidative stress markers such as MDA, GSH, SOD, CAT, GPx, and molecular endpoints are needed to clarify pathways and translational relevance.
Heavy metal contamination of industrial soils severely constrains crop productivity and environmental sustainability. This study evaluated effectiveness of algal-derived biopolymer carrageenan in mitigating multi-metal (Cd, Pb, Zn, Cu) toxicity and improving maize performance in naturally contaminated soils from four industrial zones of Pakistan (Kasur, Faisalabad, Sialkot, and Korangi). A pot experiment with graded carrageenan levels (0-200 mg kg-1) revealed significant improvements in soil physicochemical properties, including increases in pH (≈10-11%) and electrical conductivity (61-94%), alongside enhanced enzymatic activities. Carrageenan application markedly reduced DTPA-extractable metals, with maximum declines of up to 83% (Cd), 70% (Zn), 69% (Pb), and 62% (Cu), thereby limiting their bioavailability. Consequently, metal accumulation in maize roots and shoots decreased substantially, with reductions reaching 90% for Cd and 74% for Pb in shoots. These changes were accompanied by improved plant tolerance indices and significant biomass enhancement. Carrageenan also stimulated antioxidant defense systems, increasing SOD (66-79%), POD (74-150%), and CAT (56-74%) activities, along with glyoxalase function (up to 160%), while elevating non-enzymatic antioxidants such as ascorbate (up to 140%), glutathione (110%), and phytochelatins (179%). Photosynthetic pigments were significantly improved, with total chlorophyll and carotenoids increasing by up to 65% and 168%, respectively. Overall, carrageenan effectively immobilized toxic metals, reduced their translocation, and enhanced physiological and biochemical resilience in maize, demonstrating strong potential as a sustainable amendment for phytoremediation and crop production in metal contaminated soils.
Vasculogenic mimicry (VM) contributes significantly to tumor aggressiveness and resistance to anti-angiogenic therapies. Simultaneous inhibition of both angiogenesis and VM represents a promising strategy to improve therapeutic outcomes in aggressive cancers, such as triple-negative breast cancer (TNBC), which responds poorly to anti-angiogenic therapies. In this study, we identified carvacrol, a natural monoterpenoid phenol widely used as a food additive, as a dual inhibitor of angiogenesis and VM in TNBC. Carvacrol preferentially inhibited angiogenesis in endothelial cells (ECs) and VM in TNBC cells at concentrations that had minimal effects on TNBC cell proliferation. Mechanistically, carvacrol directly bound to the vanilloid-like (VL) site of transient receptor potential melastatin 7 (TRPM7), thereby inhibiting channel activity and attenuating Zn2+ influx. This triggered dephosphorylation of the mammalian target of rapamycin (mTOR) and subsequent proteasomal and lysosomal degradation of key receptor tyrosine kinases (RTKs), including vascular endothelial growth factor receptor 2 (VEGFR2), Tie2, fibroblast growth factor receptor 1 (FGFR1), and insulin-like growth factor 1 receptor (IGF1R) in ECs, as well as FGFR1 and IGF1R in TNBC cells. Genetic knockdown of TRPM7 recapitulated the anti-vascular effects and signaling alterations induced by carvacrol. In vivo, carvacrol effectively suppressed TNBC vascularization and growth in a mouse dorsal skinfold chamber model and an orthotopic xenograft model. Together, these findings suggest that carvacrol preferentially targets angiogenesis and VM in TNBC by suppressing the TRPM7/Zn2+/mTOR/RTKs axis, highlighting it as a promising therapeutic candidate for TNBC and potentially other tumors resistant to anti-angiogenic therapies, while positioning the TRPM7 channel as a novel anti-vascular target for TNBC treatment.
Hydroxysafflor Yellow A (HSYA), the major bioactive component from Carthamus tinctorius L., exerts significant protective effects against myocardial ischemia-reperfusion injury (MIRI). Mitophagy is pivotal in the pathological process of MIRI, yet the specific molecular mechanism underlying HSYA-mediated mitophagy regulation remains unclear. This study aimed to investigate the association between HSYA treatment and mitochondrial autophagy in murine MIRI and to explore the potential mechanistic role of the SIRT1-FOXO3-BNIP3 signaling pathway using functional loss-of-function and rescue experiments. These findings may provide preliminary evidence supporting the clinical translational potential in MIRI therapy. Mouse myocardial ischemia-reperfusion injury (MIRI) model and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced AC16 cardiomyocyte injury models were established. Metabolomics, molecular docking, and surface plasmon resonance (SPR) techniques were combined to screen the potential targets of HSYA. The SIRT1 inhibitor EX527 and SIRT1 siRNA were used to verify the underlying mechanism. Cardiac function, myocardial infarct size, mitochondrial function, the expression of autophagy-related proteins, and protein-protein interaction were detected and analyzed. Compared with the MIRI group, HSYA significantly improved cardiac function in mice, as evidenced by increased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (p < 0.01), attenuated ST-segment elevation, and improved myocardial perfusion. HSYA also markedly reduced myocardial infarct size (p < 0.01) and serum levels of CK-MB, LDH, and cTnI (all p < 0.01) and ameliorated myocardial histopathological damage and mitochondrial ultrastructural integrity. Mechanistic studies revealed that HSYA significantly upregulated the expression of SIRT1, FOXO3, BNIP3, Beclin-1, and the LC3II/I ratio while downregulating p62 expression (p < 0.01), consistent with enhanced mitophagy-related activity. Furthermore, these protective effects were markedly attenuated upon SIRT1 inhibition or siRNA-mediated silencing, whereas HSYA intervention partially reversed these alterations. Additionally, co-immunoprecipitation (Co-IP) and pull-down assays demonstrated that HSYA promoted protein-protein interactions between SIRT1-FOXO3, FOXO3-BNIP3, and BNIP3-LC3B. These findings highlight that HSYA is associated with improved cardiac function, enhanced mitophagy-related activity, and upregulated SIRT1-FOXO3-BNIP3 signaling, providing robust experimental evidence for its clinical translational application in MIRI treatment.
Overweight and obesity are chronic diseases that result from complex interactions including genetics, environment, eating behaviors, and limited access to a healthy diet. Amaranth protein (AmProt) has several health benefits, but no studies have examined its effects on the modulation of children's gut microbiota. The work aimed to analyze serum levels and changes in gut microbiota in children aged 8-10 years with different body mass index (BMI) values after supplementation with AmProt. Participating children were allocated into three groups according to their BMI: normal weight (NW), overweight (OW), and with obesity (OB). Children received AmProt for 90 days. Levels of fasting blood glucose, cholesterol, triglycerides, and insulin were analyzed before and after diet supplementation. HOMA-IR and adinopectin/leptin ratio were evaluated. Feces were collected and metagenome analysis was carried out. No changes in glucose levels were observed across groups and treatments; however, cholesterol and triglycerides levels tended to decrease. The HOMA-IR value increased in relation to BMI and no changes were observed after treatment. Firmicutes were highly abundant in all groups. The lower abundance of Ruminococcus was observed in the OW and OB groups. In the OW group, Blautia, Butyricicoccus, and Roseburia were also observed in increased abundance. In all groups, AmProt consumption tended to increase the abundance of Coproccus, Prevotella, and Collinsella. Conclusions: Supplementation of the children's diet with AmProt showed an improvement in serum cholesterol and triglyceride levels, which could be related to changes in the microbiota related to lipid metabolism.