Search Results (235)

The gut–immune–metabolic axis has emerged as a central regulator of human health, with growing evidence indicating that microbiota-derived metabolites improve gut microbial ecology, enhance intestinal barrier integrity, reduce systemic inflammation, and maintain metabolic homeostasis. This review synthesizes current mechanistic and clinical evidence on the role of postbiotics in regulating intestinal barrier integrity, immune responses, oxidative stress, and metabolic–endocrine homeostasis. The literature was identified through the PubMed/MEDLINE, Scopus, and Web of Science, integrating evidence from experimental, mechanistic, animal and clinical studies on the therapeutic potential of postbiotics to modulate the gut–immune–metabolic axis. Preclinical studies suggest that postbiotics may enhance epithelial barrier function by improving tight junction integrity through multiple pathways such as PI3K/Akt signaling, stimulating mucin-2 (MUC2) production, and reducing intestinal permeability. They modulate immune responses through interactions with Toll-like receptors, nucleotide-binding oligomerization domain receptors, and G-protein-coupled receptors (GPR41/43), influencing key signaling pathways, including NF-κB and Nrf2, and altering cytokine profiles, such as IL-10, TNF-α, and IFN-γ. Similarly, preclinical investigations have demonstrated that short-chain fatty acids (SCFAs) and other microbial metabolites may improve insulin sensitivity, regulate hepatic gluconeogenesis, stimulate glucagon-like peptide 1 (GLP-1) secretion, and modulate lipid metabolism through the FXR and TGR5 signaling pathways. Emerging human studies suggest potential benefits of postbiotics in regulating gut, immune, and metabolic health; nevertheless, clinical evidence remains limited and is influenced by variability in postbiotic composition, dosage, formulation, and metabolite profiles. Therefore, standardized production approaches and well-designed large-scale randomized clinical trials are required to confirm therapeutic efficacy and establish evidence-based applications of postbiotics. Full article

Inflammatory bowel disease (IBD) is characterized by chronic inflammation of the gastrointestinal tract. Current treatments, including anti-inflammatory drugs and biologics, often have limited efficacy and significant side effects, highlighting the need for novel therapeutic approaches. Phellodendrine (PHE) is a characteristic ingredient of Phellodendri chinensis, yet its effects and mechanisms on IBD remain elusive. The present study evaluated the potential of PHE for preventing dextran sulfate sodium-induced IBD in zebrafish. PHE effectively reduced inflammatory cell infiltration and modulated polarized macrophages. The qPCR results further confirmed the down-regulation of pro-inflammatory genes and up-regulation of anti-inflammatory factors. Consequently, PHE promoted the resolution of IBD inflammation. PHE also restored intestinal barrier integrity by enhancing MUC2 expression, increasing goblet cell counts, and reducing intestinal permeability of both chemical and physical barriers. In addition, PHE was associated with alterations in the gut microbiome, including a reduction in potentially pathogenic microbes and an increase in beneficial microbial populations. PHE also alleviated oxidative stress. Network pharmacology suggested the potential involvement of the IL-17 signaling pathway, the lipid and atherosclerosis pathway, and the TNF signaling pathway in the preventive effects of PHE against intestinal inflammation in the zebrafish model. In vivo gene expression analysis suggested that JUN, PTGS2, IL1B, DRD2, CALM1, and HSP90AA1 may serve as putative targets of PHE. Collectively, our results indicate that PHE demonstrates potential anti-inflammatory and barrier-protective activities in a zebrafish model of intestinal inflammation. The pharmacological mechanisms by which PHE restores intestinal barriers (microbial, chemical, physical, and immune barriers) include resolving inflammation, decreasing ROS production, and enhancing lipid accumulation in the lumen overlying the intestinal mucus barrier. This study provides novel insights into the preventive effects of PHE against intestinal inflammation in a zebrafish model, suggesting its potential as a candidate for further investigation. Full article

Pulmonary fibrosis (PF), particularly idiopathic pulmonary fibrosis (IPF), is a progressive and often fatal interstitial lung disease characterised by complex genetic and molecular heterogeneity. Traditional diagnostic approaches, which rely on clinical, radiological and histopathological assessment, are frequently insufficient to capture the underlying biological diversity of the disease. The advent of next-generation sequencing (NGS) has substantially advanced the understanding of PF by enabling comprehensive genomic and transcriptomic profiling. NGS technologies, including whole-exome sequencing (WES), whole-genome sequencing (WGS), RNA sequencing (RNA-seq), and targeted gene panels, have uncovered key genetic determinants. These include mutations in telomere-related genes (TERT, TERC, RTEL1) and surfactant-related genes (SFTPC, SFTPA2), as well as common variants like the MUC5B promoter polymorphism. These discoveries have clarified disease pathogenesis, revealed polygenic risk models, and may improve diagnostic accuracy, particularly in distinguishing overlapping interstitial lung disease (ILD) phenotypes. Beyond genetics, transcriptomic analyses have identified dysregulated pathways, including TGF-β, Wnt/β-catenin, and PI3K/Akt signalling, and have enabled the discovery of novel biomarkers for prognosis and therapeutic response. In selected clinical settings, NGS is beginning to support patient stratification and inform management decisions. Emerging applications, including liquid biopsy and integration with artificial intelligence, further expand the potential clinical utility of NGS. Despite challenges related to cost, data interpretation and standardisation, NGS represents a powerful research tool in PF. Full article

The MUCIN 1 (MUC1) gene evolved in eutherian mammals in association with the marked expansion of endogenous retroviruses (ERVs). MUC1 encodes the MUC1-C/M1C protein that protects barrier epithelia from exogenous viruses. Activation of M1C in response to loss of homeostasis induces STAT1 and the type I interferon (IFN-I) pathway. Studies in cancer cells have found that M1C regulates human ERV (HERV) expression by a STAT1-mediated mechanism. These discoveries have uncovered new insights into M1C-induced regulation of HERVs and other retrotransposons, such as LINE-1 (L1) and Alu. M1C signaling integrates retrotransposon transcription with induction of the counteracting apolipoprotein B mRNA-editing catalytic 3 (APOBEC3) genes that, like MUC1, first appeared in placental mammals. Activation of retrotransposons induces viral mimicry characterized as an IFN-I response that promotes innate anti-tumor immunity. Conversely, M1C protects cancer cells by sustained induction of the IFN-I pathway and immune evasion. This review posits that M1C-dependent regulation of retrotransposon and APOBEC3 expression represents an adaptive response exploited by cancer cells that promotes malignant progression. Full article

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a major heat-induced contaminant in protein-rich foods, yet its effects on intestinal barrier homeostasis and luminal microecology remain insufficiently defined. In this study, adult zebrafish were exposed to dietary PhIP for 90 days at estimated intake doses of 0.006, 0.4, and 7.2 mg/kg bw/day to evaluate intestinal injury, microbial dysbiosis, and metabolic remodeling. PhIP exposure impaired growth-related indices and induced progressive intestinal lesions, accompanied by mucus barrier depletion, reduced goblet cell abundance, and downregulation of muc2. Tight junction integrity was disrupted, as indicated by decreased zo-1, occludin, and claudin1 expression, weakened ZO-1 and Claudin-1 immunofluorescence signals, and reduced tight junction-related protein levels. Serum LPS and intestinal pro-inflammatory cytokines were markedly elevated, whereas il-10 expression was suppressed, indicating increased endotoxin burden and inflammatory activation. 16S rRNA gene sequencing revealed Proteobacteria-enriched dysbiosis and exposure-associated shifts in candidate genera, including Chitinilyticum, Shewanella, Aeromonas, Acinetobacter, Microbacterium, and Reyranella. Untargeted metabolomics further identified luminal metabolic remodeling involving lipid-related compounds, organic acids, amino acid metabolism, arachidonic acid metabolism, the citrate cycle, and pathways related to choline and glycerophospholipid metabolism. Association analysis linked genus-level microbial variation and core pathway-related metabolites with LPS, inflammatory cytokines, and tight junction markers. These findings indicate that dietary PhIP exposure disrupts intestinal barrier homeostasis in parallel with Proteobacteria-related dysbiosis and luminal metabolic remodeling, providing an integrated microbiota-metabolite-barrier association framework for evaluating intestinal risks of heat-induced food contaminants. Full article

Background: Chronic Obstructive Pulmonary Disease (COPD) is a progressive, incurable condition marked by irreversible airflow limitation and systemic inflammation. Cardiovascular comorbidities, particularly pulmonary hypertension (PH), exacerbate disease severity. While cigarette smoke is a well-known trigger, non-smoking-related inflammatory pathways remain underexplored. This study investigates vascular remodeling in a murine model of inflammation induced by chronic exposure to house dust mite Farinae (HDM). Methods: Female C57BL/6 mice were sensitized with HDM in Freund’s Complete Adjuvant and challenged intranasally with HDM for six weeks. Lung inflammation, mucus hypersecretion, and vascular remodeling were evaluated via BAL, histology, immunofluorescence, echocardiography, gene expression, proteomics, and FlexiVent pulmonary function tests (FlexiVent system). Results: HDM exposure induced a mixed inflammatory response, with elevated neutrophils, monocytes, and lymphocytes in BALF. Mucus hyperproduction (increase in MUC5AC/MUC5B) and impaired lung function (reduced FEV0.1/FVC) were observed. Vascular remodeling was evidenced by increased wall thickness, α-SMA expression, and collagen deposition. Proteomic analysis revealed dysregulation of endothelial markers and protease/antiprotease imbalance. HIF1-α was significantly upregulated in lung tissue and correlated with vascular and epithelial remodeling. Conclusions: Chronic HDM exposure in mice recapitulates key features observed in subsets of COPD and PH, including inflammation-driven airway and vascular remodeling. HIF1-α emerges as a central regulator, linking hypoxia to structural changes. This model offers insights into the effect of non-smoking-related inflammatory pathways on bronchial and vascular remodeling that are potentially relevant for subgroups of COPD patients and highlights HIF1-α as a potential therapeutic target. Full article

This study aimed to evaluate the protective effects of sodium humate (HNa) alone and in combination with low-dose cefixime (CFM) in mice infected with enterotoxigenic Escherichia coli (ETEC). An ETEC infection mouse model was established to compare the effects of individual or combined interventions on physiological parameters, intestinal morphology, barrier function, levels of specific intestinal bacterial groups, cell proliferation/apoptosis, and inflammatory pathways. The results showed that the HNa + CFM combination significantly promoted body weight recovery, ameliorated damage to jejunal villus structure and ultrastructure, and increased the mRNA expression of mucins (MUC1/2/3) and tight junction proteins (ZO-1, Occludin, Claudin-1) compared to the ETEC group. Concurrently, the combined treatment significantly reduced fecal E. coli counts and increased the abundance of Lactobacillus and Bifidobacterium, promoted epithelial repair by upregulating proliferation-related genes (EGFR, PCNA, TGF-β1), and decreased the Bax/Bcl-2 ratio. Furthermore, the combined intervention significantly reduced serum LPS levels and consequently suppressed ETEC-induced activation of the TLR4/MyD88/NF-κB pathway, as evidenced by reduced protein expression of TLR4 and MyD88, decreased phosphorylation of IκBα and p65, and diminished nuclear accumulation of NF-κB p65, leading to downregulation of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and elevation of IL-10. In conclusion, the combined application of HNa and low-dose CFM showed additional protective benefits against ETEC infection. These effects were associated with multi-targeted repair of the intestinal barrier, modulation of measured bacterial levels, and suppression of excessive inflammatory responses. This strategy offers a potential approach for the clinical management of bacterial enteritis and reducing antibiotic dependence. Full article

Introduction: This study sought to explore the impact of dietary Cu deficiency on colonic health, including assessments of histopathology, barrier function, inflammatory response, and gut microbiota composition. Methods: Weaned mice were fed a copper-deficient diet for four weeks, followed by one week of intraperitoneal copper sulfate administration as a proof-of-concept rescue intervention. Colonic pathology was assessed by H&E staining, goblet cell changes by AB-PAS staining, and intestinal barrier integrity by immunofluorescence. Inflammatory cytokine levels were measured by ELISA, while protein and mRNA expression of inflammatory markers were detected by Western blot and qRT-PCR. Gut microbiota composition, diversity, and signature genus abundance were analyzed by 16S sequencing. Results: Compared to the control group, CuD mice exhibited histopathological damage in the colon, including mucosal thinning and inflammatory cell infiltration. The number of goblet cells and the expression of mucin MUC2 were significantly reduced, and the expression of tight junction proteins (ZO-1, Occludin) was downregulated, indicating impairment of both the physical and chemical intestinal barriers. Concurrently, Cu deficiency markedly elevated systemic and colonic levels of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6), and enhanced NF-κB phosphorylation. To explore potential microbial contributions to these colonic alterations, we subsequently analyzed the gut microbiota composition by 16S rRNA sequencing. This analysis revealed that Cu deficiency significantly reduced the α-diversity and species richness of the gut microbiota. This dysbiosis was characterized by a decreased abundance of beneficial bacteria (e.g., Bacteroidota, Muribaculaceae) and an increased abundance of Desulfobacterota, a pro-inflammatory taxon, as well as Akkermansia, a mucin-degrading bacterium with context-dependent effects on gut health. Intraperitoneal administration of copper sulfate (CuD + CuSO₄) partially reversed the histopathological and inflammatory changes; its effect on the gut microbiota was not assessed. Conclusions: Dietary Cu deficiency is associated with colonic injury, and these alterations were accompanied by intestinal barrier disruption, an activated inflammatory response, and gut microbiota dysbiosis. These findings provide experimental evidence highlighting the importance of copper nutrition in maintaining colonic homeostasis, though further mechanistic studies are needed to establish causal relationships. Full article

Background: For almost two decades now, chimeric antigen receptor-natural killer cells (CAR-NK) have been investigated in pre-clinical breast cancer models, yet clinical evidence on efficacy remains scarce. This meta-analysis provides pooled evidence of pre-clinical CAR-NK effectiveness and safety in breast cancer and an overview of current clinical trials to support clinical translation. Methods: Following PRISMA guidelines and a registered protocol (PROSPERO, CRD420251131530), PubMed, Web of Science, and Scopus were searched up to 30 June 2025 for pre-clinical CAR-NK studies in breast cancer. Clinical studies were retrieved from clinicaltrials.gov and the International Clinical Trial Registry Platform (ICTRP) up to 1 March 2026. Pre-clinical studies without in vivo data or non-human CAR-NK cells were excluded. Primary outcomes were tumor burden (ratio of means, ROMs) and survival (median survival ratio, MSR). Data were analyzed in JASP™ and risk of bias (RoB) was assessed using SYRCLE’s tool. Results: Fourteen pre-clinical studies (38 CAR-NK treatment groups targeting EGFR, HER2, tissue factor, CD70, mesothelin, or folate receptor, with peripheral blood as the primary NK source, and a 5–10 million cell dose) and 11 early-phase clinical studies (targeting HER2, TROP2, PD-L1, MUC1, or NKG2D ligands under ongoing investigation) were included. In pooled pre-clinical analysis, CAR-NK significantly reduced tumor burden against untreated and unmodified/mock controls (ROM 0.311 [0.22–0.44] and 0.42 [0.33–0.53], p < 0.001, respectively). Survival was also prolonged significantly (MSR 1.47 [1.15–1.87], p = 0.010 vs. untreated; 1.30 [1.09–1.60], p = 0.007 vs. unmodified/mock NK cells). Subgroup analyses indicated improved efficacy with peripheral blood source and 5–10 M dosing. No treatment-related toxicities were reported. CAR-NK persistence was generally higher than unmodified/mock NK cells. Discussion and Conclusions: Significant heterogeneity was observed in ROM analysis which the multi-level meta-analysis configured as intra-study interventional variability. There was moderate RoB in pre-clinical studies. Published results from clinical trials remain limited, highlighting early stages of investigation. Overall, CAR-NK therapy demonstrated consistent pre-clinical efficacy and safety, supporting further translational and clinical evaluation in breast cancer. Full article

This study investigated whether the therapeutic efficacy of Chinese yam polysaccharide (CYP) against ulcerative colitis (UC) depends on an intact gut microbiota. A dextran sulfate sodium (DSS)-induced colitis mouse model was established, and one treatment group received broad-spectrum antibiotics (ABXs) before CYP administration to deplete the intestinal microbiota. CYP markedly attenuated colonic injury, reduced disease activity, and suppressed inflammatory mediators under both microbiota-intact and microbiota-depleted conditions. CYP also enhanced intestinal barrier integrity, as evidenced by reduced serum endotoxin levels and increased expression of MUC-2, Claudin-1, Occludin, and ZO-1. In addition, CYP improved hepatic antioxidant status by increasing GSH-Px and catalase activities and decreasing malondialdehyde levels. Moreover, CYP reduced the activation of the NF-κB and MAPK signaling pathways, with similar trends observed under microbiota-depleted conditions. Microbiota profiling showed that CYP partially corrected DSS-induced dysbiosis, whereas the ABX + CYP group exhibited distinct microbial patterns with enrichment of carbohydrate-related metabolic pathways predicted by PICRUSt2. Collectively, these findings suggest that CYP retains protective efficacy after antibiotic pretreatment, indicating that its effects may not be exclusively dependent on gut microbiota modulation, possibly involving direct actions on immune and intestinal epithelial cells. Full article

Background: Gastric signet-ring cell carcinoma (GSRCC) is an aggressive gastric cancer subtype with abundant mucin production and high metastatic propensity. However, scarcity of specific biomarkers has impeded clinical diagnosis and mechanistic research. This study systematically compares GSRCC and gastric adenocarcinoma (AC) to identify biomarkers and elucidate molecular basis of GSRCC’s aggressive behavior. Methods: We performed single-cell RNA sequencing (scRNA-seq) on surgically resected primary GC tissues, validating our findings using public datasets and functional experiments. Results: We identified expansion of a mucin-secreting epithelial subcluster (Mucous_muc5ac) in GSRCC, characterized by high MUC5AC, TFF1, and other prognosis-associated genes. Within this population, a MUCL3⁺ subpopulation (Cluster 1) spatially corresponded with classic signet-ring morphology, validating MUCL3 as a specific marker for these cells. Multi-omics analysis revealed that MUCL3⁺ signet-ring cells exhibit genomic instability, dedifferentiation, and enrichment of TNF-α/NF-κB, TGF-β/EMT, and hypoxia pathways, with elevated metastasis/angiogenesis gene scores and high TFF1 expression. Functional validation confirmed that TFF1 was associated with increased gastric cancer cell migration. Conclusions: Our study characterizes the MUCL3⁺ signet-ring cell subpopulation, highlighting the diagnostic utility of MUCL3 and suggesting TFF1 as a candidate for further investigation. These findings establish a foundation for advancing precision diagnosis and mechanistic understanding of GSRCC. Full article

Background: Shigellosis is an illness that affects young children all over the world and Shigella flexneri is one of the most common pathogens. The aim of this study was to investigate a potential beneficial effect of Weizmannia coagulans Long45 supplementation on feline-derived Shigella flexneri-induced colitis in mice, as well as potential mechanisms. Results: The results revealed that mice receiving fecal microbiota from diarrheic cats experienced significant weight loss, decreased survival rate, increased mRNA levels of inflammatory cytokines (TNF-α, IL-4, IFN-γ, IL-1β, IL-6, and IL-18), and increased cell apoptosis compared to the single DSS treatment. In contrast, mice that received fecal microbiota from healthy cats exhibited an increased body weight, increased mRNA level of ZO-1, claudin-3, and Muc2 and decreased apoptosis, indicating a protective effect. The 16S rDNA analysis revealed that the abundance of Shigella in the feces of diarrheic cats was significantly higher than that in healthy cats, while the abundance of Bacillus was lower. Using bacteria culture technology, 19 strains of Shigella flexneri were isolated from 27 fecal samples of diarrheic cats and a strain of Weizmannia coagulans Long45 was isolated from the feces of healthy cats. Further study showed that Weizmannia coagulans Long45 significantly alleviated pathological alterations and colonic barrier dysfunction by modulating the NF-κB and Nrf2 signaling pathways. Conclusions: Our data indicate that feline-derived Shigella flexneri may be a potential pathogen associated with diarrhea and intestinal barrier dysfunction. Weizmannia coagulans Long45, as a potential probiotic, can effectively alleviate Shigella-induced colitis by interfering with the Nrf2 and NF-κB signaling pathways. Full article

(1) Airborne fine particulate matter (PM_2.5) poses a growing threat to poultry production by impairing intestinal health, disturbing microbial balance, and reducing growth performance. Rosmarinic acid (RA), a natural polyphenol with antioxidant, anti-inflammatory, and gut microbiota-regulating properties, can effectively maintain intestinal homeostasis. To date, its protective effects against PM_2.5-induced intestinal injury in broilers remain largely unclear. This study investigated whether dietary RA supplementation mitigates intestinal damage and microbiota dysbiosis caused by PM_2.5 in broilers and explored the related mechanisms. (2) A total of 144 21-day-old broilers were randomly allocated to three groups, control (CON), PM_2.5 exposure (PM), and PM_2.5 exposure plus rosmarinic acid (RA), with six replicates of eight broilers each. (3) Results indicated that PM_2.5 exposure severely impaired growth performance, whereas dietary RA significantly increased average daily feed intake and average daily gain, decreased the feed-to-gain ratio, and elevated final body weight in broilers. RA significantly attenuated PM_2.5-induced intestinal inflammation, as evidenced by reduced expression of inflammatory cytokines (IL-6 and IFN-γ) and downregulation of key components in the TLR4 signaling pathway (TLR4, MyD88, and NF-κB). Inhaled PM_2.5 exposure impaired the intestinal epithelial barrier, marked by decreased mRNA levels of MUC2 and CLDN1 and increased caspase3 expression. Dietary RA treatment effectively restored these indicators, suggesting its role in maintaining epithelial integrity. Furthermore, RA reshaped the gut microbiota structure, altering both α- and β-diversity. Notably, RA led to a higher proportion of potentially health-promoting bacterial taxa, including Lactobacillus, V9D2013_group, and Oscillospirales, while reducing opportunistic pathogens like Shuttleworthia. (4) In conclusion, RA alleviates PM_2.5-induced intestinal inflammation, reinforces the epithelial barrier, and modulates the intestinal microbiota in broilers, likely through inhibition of the TLR4/NF-κB signaling. These findings reveal a novel mechanism by which RA mitigates pollutant-induced intestinal injury via gut microbiota modulation and TLR4/NF-κB suppression, offering new insights into the gut–lung axis in avian species. Full article

Parkinson’s disease (PD) and acute kidney injury (AKI) are two conditions with increasing prevalence and severe systemic complications and consequences. This research examines the combined neuroprotective and nephroprotective properties of three medicinal plants, Mucuna pruriens (Muc), Moringa oleifera (Mor), and Silybum marianum (SM), in a murine model of PD, AKI, and their comorbid state (PD–AKI), highlighting the role of the PI3K/AKT/mTOR and Nrf2/NF-κB signaling pathways. The mice were grouped as PD, AKI, or PD-AKI, and with or without the herbal pre-treatment, along with their respective controls. Motor impairments were assessed using the rotarod and pole climb assays. Biochemical indicators of renal function, oxidative stress markers, and inflammatory cytokines were quantified in kidney and brain tissues. Assessment of Nrf2, NF-κB, PI3K, AKT, and mTOR expression levels was performed using qRT-PCR. The AKI groups had significant renal impairment (4-fold increase in creatinine and 7.5-fold increase in BUN), oxidative stress (~5.5-fold increase), and increased cytokine levels (~1.5-fold increase), with downregulation of the PI3K/AKT/mTOR (~2-fold decrease) and Nrf2 signaling pathways (~1.8-fold decrease), alongside upregulation of NF-κB (~2.5-fold increase). The PD and PD-AKI groups exhibited significant neuroinflammation (~1.5-fold increase) and redox imbalance (~6-fold increase) in brain tissue, accompanied by motor impairments (1.6 to 4.6-fold decrease). Pre-treatment with Muc, Mor, and SM significantly ameliorated renal impairments (3.5-fold decrease in creatinine and ~5-fold decrease in BUN) and neurological deficits. These findings establish Muc, Mor, and SM extracts as potent, multi-target interventions capable of disrupting the feed–forward cycle of neuro-renal damage. Full article

Corneal alkali burn induces severe inflammation and tissue damage, leading to loss of corneal transparency and vision impairment. In this study, we evaluated the therapeutic potential of rapamycin (RAPA) compared with cyclosporine A (CsA) in a mouse model of corneal alkali burn, focusing on nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)–mediated inflammatory signaling and its impact on corneal wound healing and repair. Notably, RAPA robustly suppressed NF-κB activation, reduced infiltration of F4/80 macrophages and MPO neutrophils, and downregulated pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6. RAPA also markedly inhibited corneal neovascularization, as evidenced by decreased VEGF expression, reduced CD31 vessel formation, and suppression of Ang-2. RAPA substantially inhibited pathological fibrotic remodeling by reducing TGF-β1 expression, attenuating myofibroblast activation (α-SMA), decreasing collagen III deposition, and modulating matrix remodeling through suppression of MMP-9. Crucially, RAPA preserved epithelial barrier integrity by maintaining occludin expression, supported proper epithelial differentiation through sustained expression of CK12, and enhanced mucin layer stability by increasing MUC1 expression. It also restored tear production, reduced apoptotic cell death (TUNEL), and decreased dysregulated epithelial proliferation (Ki67). In conclusion, RAPA showed superior efficacy compared with CsA, primarily by enhancing corneal wound healing and facilitating structural and functional outcomes in the burned cornea. These findings underscore RAPA as a promising therapeutic candidate for ocular surface repair and vision restoration in extensive corneal injury. Full article