Search Results (1,015)

Owing to the multifactorial nature of inflammatory bowel disease (IBD) pathogenesis, conventional two-dimensional (2D) models inadequately recapitulate the complex in vivo microenvironment. This study sought to develop an immune-microenvironment-integrated intestinal-on-a-chip model to overcome these limitations. A microfluidic chip was engineered to co-culture intestinal epithelial (Caco-2) cells and macrophages, facilitating the simulation of IBD pathological conditions for mechanistic investigations. Following inflammatory stimulation, M0 macrophages polarized into the M1 phenotype, concomitant with the upregulation of pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β). This induction disrupted the expression of tight junction proteins (e.g., zonula occludens-1 [ZO-1]) in Caco-2 cells, thereby compromising epithelial barrier integrity. Infliximab was used as a model drug to inhibit TNF-α and modulate macrophage polarization within the chip, effectively rescuing impaired epithelial barrier integrity. This study establishes a reliable intestinal-on-a-chip model that recapitulates macrophage–epithelial interactions in IBD, providing a robust platform for elucidating the mechanisms underlying intestinal barrier dysfunction and developing targeted therapeutic strategies. Full article

Non-alcoholic fatty liver disease (NAFLD) is a primary metabolic disorder that threatens adolescent health globally, with no effective therapeutic agents currently available. Bellamya purificata is a traditional Chinese medicine categorized as "medicinal food", and polysaccharides are among its active components. However, its physicochemical structure remains poorly characterized, and no study has evaluated its effects on NAFLD. In this study, a homogeneous neutral polysaccharide, α-D-glucan (Mw = 6,412.704 kDa), was isolated from B. purificata. The structure of the polysaccharide was characterized using monosaccharide composition analysis, methylation analysis, NMR spectroscopy, and scanning electron microscopy. The backbone structure of the polysaccharide comprises →4)-α-D-Glcp-(1→ and →4,6)-α-D-Glcp-(1→, with side chains of α-D-Glcp-(1→ attached to the O-6 position of the 1→4,6)-α-D-Glcp-(1→ sugar residues. Additionally, QSPS-1D effectively reduced weight gain, hepatic lipid accumulation (TC and TG), and inflammatory responses (tnf-α and il-1β) in NAFLD zebrafish. Moreover, QSPS-1D alleviated dysbiosis by inhibiting harmful bacteria (e.g., Stenotrophomonas, Agrobacterium, and Chryseobacterium) and promoting beneficial microbiota (e.g., Rothia), which restored the Firmicutes-to-Bacteroidetes ratio. In parallel, it enhanced the expression of tight junction proteins (zo-1 and claudin-1), leading to the repair of the intestinal mucosal barrier. These findings suggest that B. purificata polysaccharides may be a potential functional food for early NAFLD intervention, with effects potentially associated with the modulation of the gut microbiota. Full article

Background: Corneal organoids derived from pluripotent stem cells have emerged as powerful tools for studying corneal development, disease modeling, and regenerative medicine applications. While previous protocols have successfully generated corneal tissue structures, there remains a need for three-dimensional models that recapitulate the complex cellular architecture and diversity of native human cornea. Methods: We developed a modified spontaneous three-dimensional corneal organoid model using human embryonic stem cells (hESCs) through an adapted Self-formed Ectoderm Autonomous Multi-zone (SEAM) protocol. hESCs were cultured as spheroids in ultra-low-binding plates under normoxic conditions and differentiated over 7–8 weeks. Organoids were characterized using immunofluorescence staining for corneal-specific markers and single-cell RNA sequencing to assess cellular composition and gene expression patterns. Results: Approximately 20% of organoids developed transparent regions characteristic of corneal tissue by day 30 of differentiation. Immunofluorescence analysis revealed spatially organized expression of corneal markers, including ZO-1 and E-cadherin in the outermost epithelial layers, P63α-positive putative limbal stem cells at the epithelial–stromal interface, vimentin-positive stromal cells in the interior, and laminin-1 deposition that suggests Bowman’s membrane formation. The organoids expressed cornea-specific keratins (K3, K12, and K15) and the master regulator PAX6 in appropriate cellular compartments. Single-cell RNA sequencing identified 18 distinct cell clusters, including three corneal epithelium subclusters with differential expression of MUC16, KRT12, and ΔNp63α, two stromal populations with distinct inflammatory profiles, and a corneal endothelium cluster. Transcriptomic analysis confirmed expression of key corneal genes, including AQP3, CDH1, multiple keratins, mucins, and extracellular matrix components (HAS2, CD34, CD44, COL8A1, and KERA). Conclusions: This three-dimensional spheroid-based putative corneal organoid model successfully recapitulates the multilayered architecture and cellular diversity of human cornea, including stratified epithelium, putative limbal stem cells, stroma, and endothelium in spatially appropriate arrangements. The model demonstrates molecular signatures consistent with native corneal tissue and provides a valuable platform for studying corneal development, disease mechanisms, and potential therapeutic applications. Future optimization to improve organoid formation efficiency and functional maturation will enhance the utility of this system for both basic research and translational medicine. Full article

Soil salinity limits ginger productivity, but the underlying molecular mechanisms remain largely unclear. The 14-3-3 proteins are conserved regulators in stress signaling. Here, we genome-wide characterized the 14-3-3 family in Zingiber officinale and examined the possible involvement of Zo14-3-3-03 in salt response. A total of 21 Zo14-3-3 genes were identified and classified into four groups with uneven chromosomal distribution. Among them, Zo14-3-3-03 was strongly salt-responsive: transcript levels increased 9.91- to 33.82-fold during 1–7 days of treatment and reached 62.47-fold in leaves at day 14. NaCl treatment elevated GUS expression driven by the Zo14-3-3-03 promoter. Virus-induced gene silencing (VIGS) of Zo14-3-3-03 resulted in silenced plants exhibiting higher malondialdehyde (up to 73.6%), lower antioxidant enzyme activities (SOD, POD, CAT, and APX: 18.9–31.9% reduction), reduced osmolytes (proline, soluble protein, sugars, and ascorbic acid: 23.2–36.2% reduction), excessive reactive oxygen species, and decreased relative water content. Several antioxidant-related genes were significantly downregulated. Protein interaction assays suggested a possible interaction with ZoSOS2, and the expression of SOS2 pathway genes was altered in silenced plants, indicating a potential link to calcium signaling and ion homeostasis. Taken together, these results suggest that Zo14-3-3-03 participates in ginger salt stress response possibly through redox balance, osmotic adjustment, and calcium-mediated pathways which would provide a basis for understanding 14-3-3-mediated stress responses and nominates Zo14-3-3-03 as a candidate requiring deeper validation for salt tolerance improvement in ginger. Nevertheless, due to limited functional validation, its role as a positive regulator and breeding target remains preliminary. Further genetic and mechanistic studies are needed to confirm causality and assess field-level applicability. Full article

This study compared intestinal polyamine metabolism and barrier function between Ningxiang (NX) and Duroc × Landrace × Yorkshire (DLY) piglets under baseline conditions and following ETEC challenge. Experiment 1 (baseline, n = 12/breed) assessed colonic barrier integrity, immune status, polyamines, and microbiota. Experiment 2 (ETEC challenge, n = 8/group/breed) evaluated responses to oral ETEC (10⁹ CFU) over 3 days. Under baseline conditions, NX piglets showed superior barrier integrity, higher goblet cell numbers and mucin 2 (MUC2) protein expression, and lower plasma levels of intestinal permeability markers—diamine oxidase (DAO), D-lactate (DLA), and endotoxin (ET)—compared with DLY piglets. NX piglets also exhibited reduced colonic pro-inflammatory cytokine levels (IL-6 and IL-1β) and higher expression of immune-related markers (CD3, CD68, and IgA) versus DLY piglets. In contrast, DLY piglets displayed more active microbial polyamine metabolism in the colon, with higher concentrations of putrescine, spermidine, and spermine, as well as increased ornithine decarboxylase (ODC) expression. 16S rRNA sequencing revealed greater microbial diversity and enrichment of taxa (Muribaculaceae_unclassified, Prevotella) in NX piglets, whereas DLY piglets showed enrichment of polyamine-associated genera (Collinsella, Veillonella). Following the ETEC challenge, DLY piglets displayed pronounced polyamine upregulation, including elevated polyamine levels and ODC1 expression. Conversely, NX piglets maintained more stable polyamine metabolism, higher expression of tight junction proteins (ZO-1 and occludin), lower plasma permeability markers, reduced pro-inflammatory cytokine expression (IL-6, IL-1β, IL-22), and increased anti-inflammatory IL-10 expression. Collectively, these findings demonstrate that NX piglets possess superior intestinal barrier integrity and immune maturity, while DLY piglets exhibit a more active but stress-responsive polyamine metabolic phenotype. The divergent metabolic and immune responses to ETEC challenge underscore the distinct strategies employed by these two breeds in maintaining gut homeostasis. These findings provide preliminary insights that may inform future breeding strategies aimed at enhancing intestinal health and disease resistance in pigs, pending validation in broader genetic backgrounds and mechanistic studies. Full article

Background/Objectives: The worldwide occurrence of ulcerative colitis (UC) is increasing, but existing treatments frequently suffer from limited effectiveness and notable side effects. walnut green husk polysaccharide (WGHP) has been shown to exhibit anti-inflammatory and immunomodulatory activities; however, its specific potential and mechanisms of action against colitis remain unclear. This study aimed to evaluate the effectiveness of purified WGHP on (dextran sulfate sodium) DSS-induced UC and elucidate the underlying mechanisms. Methods: WGHP-2-2, a primary acidic polysaccharide fraction, was extracted from crude WGHP and analyzed through chromatography and spectroscopy. The therapeutic efficacy of WGHP-2 was assessed using a murine model of DSS-induced UC. Assessments included disease severity (DAI, colon length, histopathology), inflammatory markers (tissue IL-6, TNF-α, IL-10), and intestinal barrier integrity (Claudin-5, Occludin, ZO-1). Results: WGHP-2-2 is an acidic polysaccharide with a molecular weight of 15.29 kDa. Its composition includes glucosamine, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, and arabinose, with respective molar ratios of 0.55, 8.48, 3.06, 65.99, 4.49, 10.86, and 6.57. Methylation and NMR analyses revealed a backbone mainly composed of →4)-α-D-GalpA-(1→ and →2)-α-D-Rhap-(1→ linkages, with side chains or terminal residues such as T-Rhap, T-Galp, T-Glcp, and T-Araf. In vivo, WGHP-2 significantly mitigated DSS-induced UC symptoms in a dose-dependent manner. Specifically, the high-dose group (123 mg/kg) markedly attenuated colon shortening and improved histological architecture, including the restoration of colonic crypts. WGHP-2 effectively reduced pro-inflammatory cytokines IL-6 and TNF-α in colon tissues, while increasing the anti-inflammatory cytokine IL-10. Conclusions: WGHP-2 mitigates DSS-induced UC by inhibiting pro-inflammatory cytokines (IL-6, TNF-α), increasing IL-10 levels, and improving intestinal barrier integrity through the upregulation of tight junction proteins. These results position WGHP-2 as a promising lead compound for developing functional foods for UC. Full article

Background: Intracerebral hemorrhage (ICH) is a devastating subtype of stroke, in which neuroinflammation and blood–brain barrier (BBB) disruption are secondary pathophysiological events that drive progressive brain injury. Histone lysine demethylase JMJD3 (Jumonji C domain-containing protein 3) is a master epigenetic switch governing inflammatory signaling; however, its participation in ICH-induced vascular disruption and its possible mechanism remain elusive. Objective: To examine the expression patterns of JMJD3 in the context of ICH and to evaluate the therapeutic potential of its specific inhibitor, GSK-J4, in attenuating neuroinflammation and BBB disruption in a murine ICH model. Methods: Hemin treatment of a mouse C8-D1A astrocytic cell line was used to develop an in vitro ICH model. The transcript level of the Jmjd3 gene and its correlation with pro-inflammatory signaling were analyzed with or without GSK-J4 pretreatment. ICH in vivo was created experimentally in adult male C57BL/6 mice through stereotactic striatal injection of collagenase IV, and the mice were randomly assigned to sham, ICH + vehicle, and ICH + GSK-J4 (30 mg/kg intraperitoneally (i.p.), every other day starting three days before ICH) groups. At three days post-ICH, ipsilateral brain tissues were collected to detect JMJD3 cellular localization, pro-inflammatory mediator levels, tight junction protein expression, BBB ultrastructure, and hematoma volume. White matter integrity and neuronal recovery were assessed on day 7, and sensorimotor function was assessed longitudinally on days 1, 3, 5, 7, and 14. Results: Jmjd3 gene transcription was upregulated in hemin-treated astrocytes and correlated positively with IL-6 pro-inflammatory signaling activation. In vivo, the co-localization of JMJD3 with the astrocytic identifier glial fibrillary acidic protein (GFAP) was markedly increased in the area adjacent to the hematoma at three days post-ICH. GSK-J4 administration significantly suppressed the pro-inflammatory signaling cascade by decreasing the levels of inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), and matrix metalloproteinase-9 (MMP-9), enhanced brain vascular structural and functional integrity by upregulating tight junction proteins zonula occludens protein-1 (ZO-1) and claudin-5, improved BBB ultrastructural integrity, and decreased hematoma volume at three days post-ICH. Furthermore, GSK-J4 administration promoted white matter integrity (increased myelin basic protein [MBP] expression) and neuronal recovery (increased neuron-specific nuclear protein [NeuN] expression) at seven days post-ICH and significantly improved the performance of ICH mice in sensorimotor behavioral tests. Conclusions: Astrocytic JMJD3 is upregulated following ICH and promotes neuroinflammation, which in turn mediates BBB disruption. Pharmacological inhibition of JMJD3 by GSK-J4 attenuates neuroinflammation and subsequent BBB damage, accelerates hematoma resolution, and promotes histological and functional recovery after ICH, likely by downregulating MMP-9 expression. These findings identify astrocytic JMJD3 as a novel epigenetic therapeutic target for acute ICH. Full article

Mastitis is a common disease in dairy cows, mainly caused by Staphylococcus aureus and Escherichia coli. Berberine (BBR) has antibacterial and anti-inflammatory potential, but its application is limited due to poor oral absorption and difficulty in reaching mammary tissue. To address this, this study developed a BBR-loaded composite ethosome hydrogel (BBR-CEH) to achieve targeted mammary delivery through local transdermal administration. The experimental results showed that BBR-CEH has good chemical stability and biosafety. Subsequently, a mouse mastitis model was established by intraductal injection of 50 µL of bacterial mixture (E. coli:S. aureus = 1:1, each at 1 × 10⁷ CFU/mL). The results showed that after BBR-CEH treatment, the mRNA expression of TNF-α (tumor necrosis factor-alpha), IL-6 (interleukin-6), and IL-1β (interleukin-1 beta) was significantly decreased, the mRNA expression of ZO-1 (zonula occludens-1), Occludin, and Claudin-4 was significantly increased, and Bax/Bcl-2 (Bcl-2-associated X protein/B-cell lymphoma 2) was significantly reduced (p < 0.01), indicating alleviation of mastitis by reducing inflammation, improving tight junctions, and inhibiting apoptosis. Finally, network pharmacology and in vivo experiments confirmed that its mechanism involves the NF-κB (nuclear factor kappa-B) and PI3K/Akt (phosphoinositide 3-kinase/protein kinase B) pathways. Thus, topical BBR-CEH may represent a promising new strategy for mastitis treatment. Full article

Chemotherapy-induced gonadotoxicity severely compromises male fertility, yet effective interventions remain limited. Building on our previous finding that yam protein (YP) modulates the gut-microbiota axis, this study investigated its direct protective role against cyclophosphamide (CTX)-induced testicular injury. Spectral analysis revealed a protein fraction (L-YP) with strong intrinsic fluorescence and optimal cytoprotection against oxidative stress. Proteomic characterization revealed six dominant proteins (YP1–YP6). In vivo experiments demonstrated that L-YP upregulates the expression of tight junction proteins Occludin and ZO-1, restores hormone levels, and modulates inflammatory factors, thereby enhancing the integrity of the blood–testis barrier. Network pharmacology analysis and molecular docking predicted a potential binding affinity between key components such as YP2 and NF-κB p65, which may provide a structural basis for their regulatory role. Further validation at the gene level indicated that YP can improve the local testicular immune microenvironment by modulating the classical TLR4/MyD88/NF-κB inflammatory signaling pathway. These findings suggest that yam protein alleviates chemotherapy-induced testicular damage, potentially through barrier protection and anti-inflammatory mechanisms, indicating its promise as a dietary protective agent. Full article

Three new, previously undescribed tanzawaic acids, steckwaic acids H–J (1–3), and twenty-three known natural products (4–26) were isolated from the marine algicolous fungus Penicillium steckii SCSIO 41040. Structurally, compound 3 underwent a rare hydration reaction at the double bond of its carboxylic acid side chain. The chemical structures and stereochemistry were determined using comprehensive spectroscopic analyses, including NMR, electronic circular dichroism (ECD) calculations, and high-resolution electrospray ionization mass spectrometry (HRESIMS), and verified by literature comparison. The protective effect of tanzawaic acids on inflammatory damage to the intestinal epithelial barrier was assessed using an LPS-stimulated Caco-2/THP-1 co-culture model. Notably, immunofluorescence and Western blotting assays showed that compound 10 significantly enhanced the fluorescence signals and protein expression of ZO-1 and occludin, alleviated lipopolysaccharide (LPS)-induced intestinal barrier damage in Caco-2 cells, and contributed to the re-establishment of intestinal barrier homeostasis. Our findings demonstrate the critical role of tanzawaic acids in maintaining intestinal barrier integrity, identifying them as promising lead compounds for UC treatment. Full article

Background: This study investigated the mechanisms underlying diarrhea induced by a high-fat diet (HFD) under a state of fatigue, focusing on gut microbiota dysbiosis, bile acid metabolic disturbance, and gut–liver injury. Methods: Mice were assigned to a normal control diet (NCD) group, a HFD-induced diarrhea under fatigue (HFDM) group, and a HFD-induced diarrhea with aggravated dysbiosis (HFDMA) group. Histopathology, inflammatory factors, intestinal barrier-related proteins, small-intestinal microbiota, and colonic bile acid profiles were assessed, and correlation analyses were performed among gut microbiota, bile acids, and inflammatory factors. Results: Compared with the NCD group, both the HFDM and HFDMA groups showed diarrhea-like and fatigue-like phenotypes, histopathological injury in the small intestine and liver, increased tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) levels, and impaired intestinal barrier function. No significant differences in inflammatory factors were observed between the HFDM and HFDMA groups. Zonula occludens-1 (ZO-1) expression decreased in both model groups but reached statistical significance only in the HFDMA group, whereas Claudin-1 expression was significantly reduced in both groups. Gut microbiota analysis showed altered community structure, with downward trends in alpha diversity that did not reach statistical significance but clear separation trends in beta diversity. Proteobacteria and Streptococcus increased, whereas Ligilactobacillus decreased. Total bile acid levels did not differ significantly among groups; however, the ratio of secondary to primary bile acids was significantly reduced in both model groups, particularly in the HFDMA group, with decreases in representative secondary bile acids, including hyodeoxycholic acid (HDCA) and isolithocholic acid (isoLCA). Correlation analysis further supported close associations among gut microbial alteration, bile acid disturbance, and intestinal and hepatic inflammation. Conclusions: Gut microbiota dysbiosis may disrupt bile acid metabolism, impair intestinal barrier integrity, and promote intestinal and hepatic inflammatory responses, thereby contributing to diarrhea progression under fatigue and HFD conditions through the gut–liver axis. Full article

Mechanical stress has been implicated in retinal pigment epithelium (RPE) dysfunction and angiogenic signaling in retinal disorders; however, its direct in vivo effects on the RPE–choroid complex remain incompletely understood. Here, we established a mouse model of localized mechanical stress by subconjunctival implantation of glass beads (0.8–1.2 mm in diameter) in eight-week-old C57BL/6J mice to induce transscleral stretching of the RPE. Ocular tissues were analyzed two days after implantation using histological, immunohistochemical, and molecular approaches, and inflammatory mediators were quantified by multiplex cytokine assays. Mechanical stress induced focal serous retinal detachment, elongation of photoreceptor outer segments, and disruption of the RPE tight junction protein ZO-1. VEGF expression in the RPE–choroid complex was significantly upregulated and accompanied by increased levels of inflammatory mediators, including MCP-1. Intravitreal administration of anti-VEGF agents effectively suppressed stress-induced VEGF expression. These findings indicate that mechanical stress is sufficient to induce structural disruption and angiogenic signaling in the RPE in vivo, providing a useful experimental platform for investigating stress-related retinal responses and therapeutic modulation of VEGF signaling. Full article

The efficacy of postbiotics varies significantly between different strains and preparation processes. We aimed at evaluating the effect of an innovative postbiotic product (iPB) generated through the sequential fermentation of Lacticaseibacillus rhamnosus GG and Lacticaseibacillus paracasei NPB-01, compared to single-strain postbiotics, on epithelial barrier integrity and innate immunity in human enterocytes using a Caco-2-cell-based experimental model by measuring human enterocyte proliferation and differentiation (lactase expression), tight junction proteins (occludin and zonula occludens 1, ZO-1), and mucus protein Mucin-2 (Muc-2) expression. The modulatory action on the major innate immunity peptide, Human Beta-Defensin 2 (HBD-2), production was also assessed. The iPB exposure resulted in a higher up-regulation of human enterocyte proliferation and differentiation, as suggested by higher lactase expression, and of occludin, ZO-1, and MUC2 expression compared with the single-strain postbiotics, suggesting a beneficial synergistic action in modulating the epithelial gut barrier. Furthermore, iPB induced a higher production of HBD-2, suggesting a synergistic enhancement of innate immune response. Our findings suggested that the sequential fermentation process could act as a biotechnological catalyst, optimizing the gut-barrier-protective properties and the immunomodulatory action of Lacticaseibacillus strains. This study introduces iPB as a high-performance postbiotic candidate for the prevention and management of conditions characterized by alterations in epithelial gut barrier and innate immunity. Full article

Antibiotic-associated diarrhea (AAD) is primarily driven by disruption of the gut microbiota accompanied by intestinal mucosal injury. Although multiherb formulations are widely used in East Asian medicine, their collective ecological effects and integrated microbiome–host mechanisms have not been systematically synthesized. This systematic review included 17 preclinical studies that investigated multiherbal formulations in AAD models. Given the substantial heterogeneity in the formulation composition, experimental design, and analytical platforms, a descriptive synthesis was performed. The included formulations were categorized into four clusters based on their shared herbal composition: Qiwei Baizhu San (QWBZP), Lizhong Tang (LZT), Gegen Qinlian Tang (GQT), and other supportive multiherbal formulations. The cluster-based synthesis revealed distinct convergent therapeutic strategies. The QWBZP and LZT clusters primarily supported the restoration of host metabolic and digestive functions, whereas the GQT cluster exhibited potent pathogen control effects with the suppression of opportunistic taxa. Across all clusters, a convergent microbiome–host response emerged, characterized by enrichment of commensal bacteria (e.g., Lactobacillus), upregulation of tight junction proteins (e.g., ZO-1, occludin), and attenuation of pro-inflammatory mediators (e.g., TNF-α, myeloperoxidase). Multiherb formulations in AAD models not only act as microbial modulators but also function as host-directed modulators that stabilize the intestinal homeostatic niche. Botanical interventions may facilitate endogenous microbiome recovery by reinforcing mucosal integrity and reducing environmental resistance. This ecological framework provides a rationale for future translational studies evaluating integrated herbal–probiotic strategies and precise microbiome management for patients with AAD, while further clinical validation is warranted. Full article

This study investigated the effects of chronic heat stress (HS) on brain injury in broilers and the associated molecular changes. A chronic HS model was established by exposing broilers to 35 °C from 08:00 to 20:00 daily from 21 to 42 days of age, and samples were collected at 28, 35, and 42 days of age. Chronic HS significantly impaired growth performance and was associated with histopathological and ultrastructural alterations in brain tissue. Serum antioxidant enzyme activities and the total antioxidant capacity were significantly reduced, whereas malondialdehyde levels were significantly increased, indicating sustained oxidative stress (OS). Blood–brain barrier (BBB) permeability, assessed by Evans blue extravasation, was significantly higher in HS birds and was accompanied by reduced mRNA expression of the tight junction-related genes ZO-1 and Claudin-5. In addition, chronic HS was associated with increased mRNA expression in genes related to cellular stress, oxidative stress, and inflammation, including key components of the TLR4/MyD88/NF-κB/NLRP3 pathway, as well as decreased expression of IL-4. These findings suggest that chronic HS is associated with enhanced OS, altered neuroinflammatory gene expression, and BBB impairment in the broiler brain. Overall, this study provides evidence that chronic HS is associated with brain injury in broilers and highlights a potential link among OS, inflammation-related transcriptional changes, and BBB dysfunction, thereby offering a basis for further mechanistic and interventional studies. Full article