Fibrosis is a pathological condition resulting from an excessive tissue response during the repair process, often affecting various tissues such as the skin, organs, and joints, posing a significant threat to global health. Researchers have made substantial efforts to explore the endogenous mechanisms underlying fibrosis in recent years and have developed several therapeutic strategies to block this process. Historically, research on fibrotic diseases has focused on identifying highly relevant therapeutic targets and developing effective antifibrotic drugs. However, due to the complexity of the mechanisms of fibrosis and its effector cells, the effectiveness of antifibrotic therapies remains limited. With the advancement of high-throughput omics technologies and machine learning tools, we now have a clearer understanding of cellular heterogeneity, intercellular interactions, and the specific roles of cells in various biological processes. This enables tracking the trajectory of different cell types during the fibrotic process, facilitating early identification and discovery of new targets for fibrosis treatment, and conducting more precise targeted research. Supported by these novel technologies, numerous studies have revealed that, in addition to normal fibroblasts, a group of bone marrow-derived fibrocytes also contributes to the fibrosis of both parenchymal and non-parenchymal organs and tissues. Circulating fibrocytes are hematopoietic-derived cells that are recruited to injury sites during injury, disease, and aging, acting as participants in inflammation and tissue repair, and directly or indirectly promoting fibrosis in various tissues throughout the body. This review summarizes the general characteristics of circulating fibrocytes, the molecular mechanisms involved in their recruitment to different tissues, the process of their differentiation into fibroblasts, their potential roles in various diseases, and the latest research developments in this field. Given the key role of circulating fibrocytes in fibrosis across multiple tissues, they may serve as promising targets for the development of novel antifibrotic therapies.

Burn injuries constitute a significant global health challenge, with deep partial-thickness burns (deep second-degree) posing particular clinical concerns due to prolonged healing and high scarring risks stemming from reticular dermis damage. Current therapeutic strategies remain largely empirical, reflecting limited understanding of stage-specific regulatory mechanisms. This study systematically investigated the molecular basis of deep partial-thickness burn repair by establishing murine models and performing RNA-seq analysis across healing phases (0, 3, 7, 14 days post-burn, dpb). Integrated bioinformatics revealed pivotal ceRNA and PPI networks, identifying hif1a (hypoxia-responsive immunomodulator) and col1a1 (ECM remodeling hub) as nodal regulators. Mechanistically, mmu-miR-101a-3p and mmu-miR-181a-5p were validated as post-transcriptional repressors of col1a1 and hif1a, respectively. Our work pioneers the discovery of the mmu-miR-181a-5p/hif1a and mmu-miR-101a-3p/col1a1 axes as master regulators of burn repair, offering novel therapeutic targets. The multi-omics dataset and molecular networks established herein provide a foundational resource for wound healing research.

Chronic wounds are a significant global health concern and remain an unresolved challenge. In recent years, with rapid advancements in materials science and the deep integration of biomedicine and materials science, numerous inflammation-modulating dermal substitutes (IMDs) that integrate anti-inflammatory biomaterials and bioactive components have emerged, providing new strategies for the treatment of chronic wounds by precisely regulating the inflammatory microenvironment. This article reviews the roles and mechanisms of actions of IMD scaffolds with varying compositions, physical properties, and drug loadings, focusing on the core pathological mechanism—the imbalanced inflammatory response in chronic wounds. It aims to provide valuable references for researchers and clinicians in related fields and to promote the further development and application of IMD scaffolds.

Burns cause skin and deep tissue damage, with ~180,000 annual deaths worldwide, mostly in developing countries. Extensive burn patients are prone to secondary lung, kidney, liver, and heart dysfunctions, and infection-induced sepsis is a major cause of mortality. With August 20, 2025 as the retrieval cutoff, we systematically searched PubMed to review burn-induced organ injury progress. This review elaborates on each organ injury's pathological mechanisms (inflammatory activation, endothelial disruption, pyroptosis, ferroptosis, etc.), clarifies post-burn cross-organ crosstalk, and summarizes emerging therapies (nano-targeted therapy, mitochondrial protection, stem cell intervention) and their clinical potential. It aims to provide a theoretical basis for post-burn organ dysfunction treatment and promote the transformation from single-organ protection to systemic intervention.

Objective·To investigate the effects of bacteria on fibroblast function in vitro and the impact of bacterial intervention on hypertrophic scars in rabbit ears in vivo.Methods·A total of 16 clinical hypertrophic scar specimens and normal skin tissues were collected from patients admitted to the Department of Burns, Plastic Surgery and Wound Repair, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, from January 2023 to December 2024, and subjected to Gram staining. After amplification and inactivation, standard Staphylococcus aureus was used to treat fibroblasts derived from normal skin at concentrations of 102, 103, and 104 CFU/mL. The effects on fibroblast proliferation were observed by using Edu staining, while α-smooth muscle actin (α-SMA) expression was detected by immunofluorescence. The expression levels of vascular endothelial growth factor (VEGF), transforming growth factor-β1 (TGF-β1), type Ⅰ collagen and type Ⅲ collagen were measured by Western blotting. Additionally, the secretion of inflammatory cytokines, including interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and monocyte chemoattractant protein-1 (MCP-1), was evaluated by using enzyme-linked immunosorbent assay (ELISA). In the in vivo study, 80 wounds were created on the ears of 10 New Zealand white rabbits, followed by inoculation with Staphylococcus aureus. After hypertrophic scar formation, intradermal injections of levofloxacin or saline were administered. Scar growth was monitored, and Vancouver Scar Scale scores were recorded. Scar tissues were harvested for hematoxylin and eosin staining, Masson staining, and Gram staining, along with immunohistochemical detection of IL-6, IL-8, TNF-α, and MCP-1 expression.Results·Clinical scar tissues exhibited significantly greater abundance of Gram-positive bacteria than normal skin tissues (P=0.001). In vitro experiments demonstrated that low-abundance bacteria (102 and 10³ CFU/mL) significantly promoted fibroblast proliferation (P=0.045, P=0.017) and α-SMA expression (P=0.042, P=0.002). These bacteria also enhanced the expression of VEGF (P=0.023, P=0.011), TGF-β1 (P=0.029, P=0.031), type Ⅰ collagen (P=0.032, P=0.025), and type Ⅲ collagen (P=0.019, P=0.027), as well as the secretion of inflammatory cytokines IL-6 (P=0.023, P=0.011), IL-8 (P=0.021, P=0.009), TNF-α (P=0.029, P=0.011), and MCP-1 (P=0.023, P=0.008). In the rabbit ear model, levofloxacin injection significantly reduced scar hyperplasia at 45 and 60 days (P=0.019, P=0.013). At 60 days, treated scars showed decreased inflammatory cell infiltration, reduced bacterial load, less collagen deposition, and lower expressions of IL-6 (P=0.025), IL-8 (P=0.021), MCP-1 (P=0.028), and TNF-α (P=0.019).Conclusions·Low-abundance bacteria promote the profibrotic capacity of scar fibroblasts, while antibiotic application effectively mitigates hypertrophic scar formation.

BACKGROUND:Sepsis-induced acute lung injury (ALI) remains a leading cause of mortality in critically ill patients, characterized by endothelial barrier dysfunction and uncontrolled inflammation. While microRNAs regulate endothelial responses during sepsis, the role of miR-149-5p remains unclear. METHODS:We conducted RNA-seq analysis to screen miR-149-5p expression in lipopolysaccharide (LPS)-stimulated endothelial cells, followed by functional validation via in vitro modulation of miR-149-5p with assessment of junction proteins (ZO-1, occludin, VE-cadherin) and adhesion molecules (ICAM1), as well as in vivo studies using a cecal ligation and puncture (CLP) mouse model treated with agomiR-149-5p. Mechanistic studies included bioinformatics predictions, dual-luciferase reporter assays to verify targets, and ABCA1 knockdown/overexpression rescue experiments. RESULTS:Our findings revealed significant downregulation of miR-149-5p in LPS-stimulated endothelial cells. Restoring miR-149-5p expression upregulated tight/adherens junction proteins (ZO-1, occludin, VE-cadherin) and suppressed ICAM1 expression. In CLP mice, agomiR-149-5p attenuated lung injury by reducing alveolar edema, protein leakage, and immune cell infiltration, and by improving endothelial barrier function. Mechanistically, dual-luciferase assays confirmed direct binding of miR-149-5p to the ABCA1. ABCA1 knockdown mimicked the protective effects of miR-149-5p, while its overexpression exacerbated barrier dysfunction; furthermore, ABCA1 restoration abolished miR-149-5p-mediated protection. CONCLUSIONS:The miR-149-5p/ABCA1 axis represents a novel regulatory mechanism of endothelial homeostasis in sepsis-induced ALI. AgomiR-149-5p delivery demonstrates therapeutic potential by restoring barrier integrity and suppressing inflammation, offering a targeted strategy for critical care management.

Patients with diabetes frequently face challenges associated with impaired wound healing, which is a serious clinical concern. Regulatory T cells (Tregs) are essential immune modulators that are integral to wound healing. While research is increasingly examining alterations in Tregs and their potential impacts on diabetic wound healing, a comprehensive review is lacking. This review employs a narrative approach to summarize the current knowledge regarding the mechanisms by which Tregs influence wound healing in diabetes, focusing on their impact on inflammatory responses, cellular regeneration, and angiogenesis. Furthermore, we investigated the potential therapeutic applications of Tregs in diabetic wound healing, highlighting recent advances in biomaterials, molecular pathways, and regulatory strategies targeting Tregs. Despite challenges such as cell acquisition and quality control, as well as risks like immunosuppression and infection, advancements in clinical studies indicate that Tregs could be a promising new treatment for diabetes. This review aims to provide novel insights and directions by analyzing the most recent literature on future therapeutic strategies for managing diabetic wounds.

BACKGROUND:This multicenter, randomized controlled trial evaluated the safety and efficacy of PermeaDerm, a biosynthetic wound matrix, compared to porcine acellular dermal matrix (ADM) for the treatment of partial-thickness burns in Chinese patients. METHODS:A total of 184 patients were enrolled and randomized 1:1 to receive either PermeaDerm or ADM across the four Chinese centers. The primary endpoint was the proportion of wounds that achieved complete re-epithelialization by day 14 (superficial partial-thickness burns) or 21 (deep partial-thickness burns). The secondary endpoints included complete re-epithelialization over time, wound area reduction rate (cm2/day), time to complete re-epithelialization, pain scores, device performance, and incidence of complications. RESULTS:PermeaDerm was non-inferior to ADM for the primary endpoint. On day 7, significantly more wounds treated with PermeaDerm achieved early complete re-epithelialization for superficial partial-thickness burns (55.6% [20/36] vs. 27.0% [10/37], p = 0.012) and deep partial-thickness burns (37.2% [19/51] vs. 12.7% [7/55], p = 0.005). The median wound area reduction rate at day 7 was also significantly higher with PermeaDerm for both superficial partial-thickness burns (2.41 cm²/day (IQR 0.86-5.07) vs. 1.05 cm²/day (IQR 0.34-3.01), p = 0.034) and deep partial-thickness burns (3.50 cm²/day (IQR 1.40-8.02) vs. 1.78 cm²/day (IQR 0.85-4.40), p = 0.021). After day 7, the healing rates were comparable between the groups. Median time to complete re-epithelialization was 3 days shorter with PermeaDerm for both superficial (7.00 days (IQR 6-10.25) vs. 10.00 days (IQR 7-12.75), p = 0.016) and deep partial-thickness burns (10.00 days (IQR 7-13) vs. 13.00 days (IQR 10-15), p = 0.003). Pain scores and complication rates were similar between the groups, while PermeaDerm was associated with more favorable ease of application, wound visibility, and removal. CONCLUSION:PermeaDerm accelerates early wound healing and shortens the time to complete re-epithelialization compared with ADM, while maintaining a comparable safety profile. These findings suggest that PermeaDerm may represent a clinically useful alternative for the management of partial-thickness burns.

The blood urea nitrogen-to-albumin ratio (BAR) has been proposed as a prognostic marker in various clinical settings, yet its predictive value in patients with extensive burns remains unclear. This study aimed to explore the potential association between the BAR and the prognosis of patients with extensive burns. We conducted a retrospective, single-center cohort study at the Burn Center of Changhai Hospital, including patients admitted between May 1, 2005, and December 31, 2022. Eligible patients were randomly assigned to a training set and a validation set in a 7:3 ratio. Univariate and multivariate Cox regression analyses were performed to identify mortality-associated factors, including age, sex, burn etiology, total body surface area burned (TBSA), full-thickness burn area (FTBA), blood urea nitrogen, serum creatinine, serum albumin, and BAR. The predictive value of BAR was assessed using receiver operating characteristic (ROC) curves and Kaplan-Meier survival analysis. The model was validated using the internal validation set. A total of 372 patients with extensive burns were included (73.92 % male; 81.99 % flame burns). Age, TBSA, FTBA, inhalation injury, tracheostomy, and BAR were identified as independent predictors of 30-day mortality. The same variables, excluding age, also predicted 60-day mortality. BAR outperformed serum albumin in prognostic accuracy. The area under the receiver operating characteristic curve for 30-day mortality was 0.801 (95 % confidence interval [95 % CI]: 0.729-0.872; sensitivity: 0.750; specificity: 0.728) and for 60-day mortality was 0.801 (95 % CI: 0.734-0.867; sensitivity: 0.714; specificity: 0.731). BAR is an accessible, cost-effective prognostic marker with high predictive accuracy for 30-day and 60-day mortality in patients with extensive burns. It demonstrates superior prognostic utility compared to individual markers and may support risk stratification in burn care.

The urgent need for silver-based antibacterial agents in clinical settings has driven the diversification of their delivery systems, evolving from traditional silver salt preparations to new silver nanoparticles (AgNPs) and silver-based composite functional materials. Research and application of various carrier systems have established a solid foundation for the clinical translation of silver. However, it is important to recognize that the clinical use of silver-based materials still faces several key challenges: one is the potential risk of cytotoxicity, another is the growing trend of bacterial resistance to silver, and the third is the heterogeneity of antibacterial properties in different wound microenvironments. Additionally, this study thoroughly examines the significant gap between basic research and clinical application of silver-based materials, highlighting that the lack of standardized clinical endpoint indicators and high-quality clinical research evidence are the main barriers to its standardized use. Future research should focus on four key areas: developing precise targeted delivery systems, creating combined treatments with silver and other antibacterial agents, enhancing biosafety through material engineering, and establishing a unified framework for clinical efficacy evaluation. Through systematic innovation and evidence-based clinical implementation, silver-based technologies hold broad potential and significant clinical value for addressing complex wound infections and alleviating the global antibiotic resistance crisis.

Skin shows distinct temporal dynamics and spatial heterogeneity during development, aging, disease, and regeneration. Although single-cell sequencing has revealed cellular diversity, its lack of spatial context limits the ability to characterize cells within their native tissue microenvironment. Factors such as acute injury and chronic wounds spatiotemporally disrupt skin homeostasis and induce complex remodeling and functional changes. Understanding these dynamic processes with spatiotemporal resolution remains a challenge in skin biology. Recent advances in spatiotemporal omics make it possible to integrate single-cell sequencing, spatial omics, and time series analyses, allowing the preservation of in situ cellular positions and revealing gene expression dynamics and intercellular networks. These technologies have reshaped the understanding of skin development and wound healing and have promoted advances in precision medicine and regenerative therapies. In this review, the applications of, recent advances in, and clinical translation potential of spatiotemporal omics in skin research are summarized. The construction of a high-resolution, spatiotemporal cellular atlas across the human skin life cycle will help identify key biomarkers, optimize regenerative strategies, and support personalized therapies.

Dendritic cells (DCs) are crucial antigen-presenting cells that mediate the interplay between innate and adaptive immunity during lethal infections. Here, we report the key role of reticulophagy regulator 1 (RETREG1), a selective autophagy receptor, in maintaining DC maturation and function in the early stage of sepsis. Mechanistically, activating transcription factor 6 (ATF6) acts as a direct transcription factor regulating RETREG1 expression in response to bacterial lipopolysaccharide-induced endoplasmic reticulum (ER) stress. RETREG1-mediated reticulophagy reduces excessive ER stress via the eukaryotic translation initiation factor 2 alpha kinase 3 (EIF2AK3) signaling pathway and inhibits membrane-associated RING-CH-type finger 8 (MARCH8)-dependent major histocompatibility complex class II (MHC-II) ubiquitination to maintain antigen presentation in DCs. Consequently, Cd11ccreRetreg1fl/fl, Retreg1-/-, and Atf6-/- mice exhibit impaired DC function, leading to immunosuppression and multiple organ failure in experimental sepsis. Exploration of samples from septic patients, combined with single-cell bioinformatics analysis, further suggests that a deficit in reticulophagy in DCs is associated with the development of human sepsis.

Objective: We aim to determine the optimal timing and approaches for first tangential excision of severely burned patients ≥70% total body surface area (TBSA). Background: Early tangential excision is the gold-standard surgical therapy for full-thickness burns. However, there are debates about its optimal timing and approaches for severely burned patients ≥70%TBSA. Materials and Methods: This study included 185 patients in the Kunshan factory explosion. We focused on surgical timing, surgical area, and main wound covering method for first tangential excision. We determined the optimal cut-off values for surgical timing and area using R language “surv_cutpoint” and “surv_categorize” functions, and utilized the Kaplan-Meier survival analysis to investigate their influences on patients’ overall survival. Then, we utilized multi-variate Cox regression analysis to identify independent factors, and performed subgroup analyses to find confounding factors. Finally, we employed a 10-year single-center cohort of 144 patients with burn injury ≥70% TBSA for external validation. Results: Performing first tangential excision in >3 days (hazard ratio=2.37, 95% confidence interval=1.05-5.40, P =0.039) and surgical area > 48% TBSA (hazard ratio=2.41, 95% confidence interval=1.46-4.00, P <0.001) were independent risk factors in the Kunshan cohort for overall survival of patients ≥70% TBSA, and were associated with higher rate of complications. Subgroup analysis revealed that the timing and TBSA of first tangential excision were influenced by several treatment and complications factors (hemodynamic status and types of hospitals). Main wound covering methods was a significant confounding factor. Patients with more autograft showed better overall survival than those with more extensive use of temporary skin substitutes in the Kaplan-Meier survival analysis ( P <0.001). However, main wound covering methods influenced by the severity of injury, availability of autologous skin source, as well as the medical resources. Additionally, external validation demonstrated consistent results and acceptable calibration. Conclusion: This study identified the optimal timing and approaches for first tangential excision, validating them as independent prognostic factors for patients with ≥70% TBSA. However, considering the various confounding factors, treatment for each patient should be tailored.

Keloids are dermal fibroproliferative skin disorders caused by abnormal wound healing, resulting in impaired skin function and aesthetic defects. Abnormal fibroblast proliferation and excessive collagen deposition are involved in keloid formation. This study investigated the role of fibroblast differentiation in keloid development. Single-cell and bulk RNA sequencing data of keloids were comprehensively analyzed, and 25 clinically relevant differentially expressed fibroblast-differentiation-related genes (DEFDRGs) were identified. Based on DEFDRGs, a keloid diagnostic classification system comprising three subtypes was constructed, indicating that DEFDRGs could serve as therapeutic targets. Additionally, multiple microarray datasets, protein sequencing data, and immunohistochemical analyses of key markers in clinical keloid samples were used for further verification. In conclusion, this study established a molecular classification of keloids based on fibroblast differentiation, contributing to the further understanding of keloid pathogenesis and providing new insights for diagnosis and treatment.

Diabetic wounds are characterized by impaired angiogenic response and poor vascular networks. Therefore, exploring the underlying mechanism and thus developing the therapeutic strategy targeting angiogenesis hold great promise in diabetic wound healing. JAM-A, a classical cell adhesion molecule, is implicated to participate multifunctional biological processes in diverse cells; however, the precise involvement of endothelial cells (ECs) JAM-A in diabetic wound healing remains unknown. Here, we found that JAM-A predominance in cutaneous endothelia was significantly reversed in diabetic environment and JAM-A restoration in ECs orchestrated angiogenic capacities and alleviated inflammatory response which were deteriorated by sustained hyperglycemic culture. An ECs-specific gene therapy based on lipid nanoparticles was further designed and resulted in ECs JAM-A restoration with significantly better angiogenesis, improved inflammatory microenvironment and accelerated wound healing in diabetes. This therapeutic effect was mainly mediated by the activation of PI3K/AKT/mTOR signaling pathway. Our data reveal that ECs JAM-A may represent a new therapeutic target in diabetic wound and potentially other diseases characterized by impaired angiogenesis.

Introduction This study focuses on the prognostic impact of ABO blood type in patients with severe burn injuries, particularly following the Kunshan explosion, exploring an under-investigated area of burn injury outcomes.Method Data from patients admitted after the Kunshan explosion were analyzed, using chi-square tests and multivariate Cox regression to investigate the relationship between ABO blood type and overall survival (OS). The study utilized various performance metrics, including the receiver operator characteristic curve (ROC), decision curve analysis (DCA) and Kaplan-Meier survival curve, for model assessment, along with a subgroup analysis for associated factors.Result The research identified ABO blood type as an independent prognostic factor in severe burn injuries. It was found that blood type O is associated with better OS, while blood type B is linked to lower OS (p = 0.001). Clinical variables such as acute kidney injury (AKI), sepsis, and other complications influenced the effect of ABO blood type on OS, according to our subgroup analysis.Conclusion Our findings indicated that blood type B was an independent and significant predictor of OS in patients with severe burn injuries, likely due to differences in immune and coagulation responses. Although the study focused on the relationship between blood type and OS, it also highlighted the importance of other factors, such as patient comorbidities and treatment approaches.

Macrophage-mediated acute inflammation is crucial for pathogen clearance and tissue repair, yet the underlying molecular mechanisms remain inadequately understood. The present study focused on the dynamic profiles of the proteome and phosphoproteome of macrophages exposed to lipopolysaccharide within 1 h. Gene Set Enrichment Analysis (GSEA) identified significantly enriched pathways in fatty acid metabolism and translation during the early inflammatory phase. Further trend analysis of the differentially expressed proteins revealed patterns associated with translation regulation such as translation initiation. Importantly, the nascent chain experiment demonstrated no significant changes in overall gene translation levels during this phase. These data indicate that macrophages maintain intracellular protein homeostasis through translational regulation, with post-translational modifications (PTMs) playing a crucial role in the rapid cellular response to pathogen invasion. Phosphorylation is a key PTM that regulates protein functions in almost all cellular processes. Time-resolved phosphoproteome analysis identified 367 differentially expressed phosphopeptides involved in immune-related pathways that resist infection. Additionally, weighted gene co-expression network analysis (WGCNA) discovered core modules that regulate translation-related processes such as RNA export from nucleus. Moreover, conjoint analysis of the proteome and phosphoproteome identified the hub protein EF1B that exhibited the largest fold change and is also involved in translation. Our data not only provide a more comprehensive understanding of the dynamic molecular networks of acute macrophage inflammation but also provide a systematic proteomic resource for further studies.

Cutaneous fibroblast heterogeneity is mechanistically linked to wound repair outcomes and fibrotic progression, with glycosphingolipid metabolism emerging as a critical determinant of physiological fibroblast diversity. Through integrative analysis of spatiotemporal omics, lipidomics, and single-cell RNA sequencing (scRNA-seq) coupled with histological evaluation of clinical specimens, the functional involvement of globotriaosylceramide (Gb3) in dermal regeneration processes is systematically investigated. Comparative profiling reveals significant upregulation of Gb3 biosynthesis in superficial second-degree burns (SSDB) relative to deep second-degree burn (DSDB) injuries. Hexosaminidase subunit beta (HEXB) is identified as the exclusive differentially expressed Gb3 synthase distinguishing these injury subtypes. Functional validation through in vitro and in vivo models demonstrates that pharmacological suppression of HEXB-mediated Gb3 synthesis exacerbates fibroblast-to-myofibroblast transdifferentiation, attenuated fibroblast growth factor 2 (FGF2) signal transduction, and ultimately potentiated fibrotic scarring. These findings establish a novel HEXB-Gb3-FGF2 regulatory axis governing fibroblast phenotypic plasticity in differential-depth skin injuries, providing mechanistic insights for developing targeted antifibrotic therapies.

Background:Sepsis-associated acute lung injury (ALI) is driven by endothelial barrier dysfunction and endothelial-mesenchymal transition (EndoMT), mediated by TGF-β1/SMAD3 signaling. Despite the therapeutic potential of SMAD3, current inhibitors face limitations. As endogenous small molecules that are closely related to physiological regulatory processes, microRNAs (miRNAs) have more potential research value for regulating SMAD3. Therefore, this study aimed to investigate the protective effect and molecular mechanism of a key miRNA targeting SMAD3 in sepsis-ALI. Methods:Screening multiple databases revealed that miR-23b-3p was the sole miRNA targeting SMAD3. Lipopolysaccharide (LPS)-stimulated human umbilical vein endothelial cells (HUVECs) and cecal ligation/puncture (CLP) mice were used to model sepsis. Lentivirus was used to construct stable strains. The functional performance and mechanism were verified by key techniques, including dual-luciferase assays, rescue experiments, reverse transcription-quantitative polymerase chain reaction (qPCR)/Western blotting, monocyte adhesion/permeability assays, and histopathology. Results:In LPS-stimulated HUVECs, miR-23b-3p downregulation correlated with TGF-β1/SMAD3 activation, EndoMT progression, and barrier disruption. miR-23b-3p overexpression reversed these effects by restoring the expression of junctional proteins and suppressing the expression of mesenchymal markers. Chromatin isolation by RNA purification-qPCR, RNA pull-down, and dual-luciferase assays confirmed the direct miR-23b-3p-SMAD3 3'UTR interaction. Rescue experiments demonstrated that miR-23b-3p counteracts TGF-β1/SMAD3 hyperactivation. In CLP mice, intratracheal agomiR-23b-3p attenuated lung injury, normalized alveolar architecture, and reduced vascular leakage by suppressing endothelial Smad3 upregulation. Conclusion:miR-23b-3p is a SMAD3-targeting regulator that inhibits EndoMT and repairs endothelial barrier integrity. Mechanistically, miR-23b-3p preserves endothelial homeostasis via SMAD3-dependent EndoMT inhibition. This study provides mechanistic insights and a miRNA-based therapeutic strategy for sepsis-induced ALI.