Mediators of Inflammation最新文献

Purpose: To investigate the therapeutic potential of recombinant thrombomodulin domain 1 (rTMD1) in diabetic corneal wound healing and to elucidate its underlying mechanisms using in vitro and in vivo models.

Methods: rTMD1 was produced using the Pichia pastoris expression system and purified. Human corneal epithelial cells (HCECs) were cultured under normal glucose (NG) and high glucose (HG) conditions, with or without rTMD1 treatment. Wound healing rates were evaluated using a scratch assay. Diabetes was induced in C57BL/6 mice via streptozotocin (STZ) injections. Corneal wounds were created and treated with rTMD1 or PBS, and wound healing was assessed via fluorescein staining. Inflammatory markers, including HMGB1, TLR4, NLRP3, and IL-1β, were analyzed via quantitative PCR (qPCR), Western blot, and immunofluorescence staining.

Results: In vitro, HCECs treated with rTMD1 under HG conditions demonstrated a higher wound healing rate compared to untreated cells (p = 0.0049). In vivo, rTMD1 significantly enhanced corneal wound healing in diabetic mice, with faster wound closure compared to PBS-treated controls at 24 h (p = 0.005) and 48 h (p < 0.0001). rTMD1 treatment reduced the expression of HMGB1, TLR4, NLRP3, and IL-1β at both mRNA and protein levels, indicating suppression of inflammation.

Conclusions: Topical application of rTMD1 promotes corneal epithelial wound healing in diabetic conditions by inhibiting HMGB1/TLR4/NLRP3-mediated inflammation. rTMD1 holds promise as a potential therapeutic agent for diabetic keratopathy, although further studies are needed to validate its clinical efficacy and safety.

Objective: Inflammation is a fundamental biological response that serves to protect the body from physical damage and harmful stimuli. However, chronic inflammation is implicated in the pathogenesis of various inflammatory diseases. Stachys pilifera Benth. has been traditionally used for its anti-inflammatory properties, yet its precise mechanisms of action remain to be fully elucidated. This study aims to investigate the effects of S. pilifera Benth. extract and its fractions on lipopolysaccharide (LPS)-induced inflammatory responses in RAW264.7 macrophage cells.

Methodology: The anti-inflammatory potential of the methanolic extract and its ethyl acetate, butanol, and water fractions were evaluated by assessing their inhibitory effects on nitric oxide (NO) and prostaglandin E2 (PGE₂) production. Additionally, nuclear factor-κB (NF-κB) concentration and cyclooxygenase-2 (COX-2) gene expression were analyzed in LPS-stimulated macrophage cells.

Results: The methanolic extract, ethyl acetate fraction, and butanol fraction significantly reduced the production of NO and PGE₂, decreased NF-κB concentration, and suppressed COX-2 mRNA expression in a dose-dependent manner. The butanol fraction exhibited the most potent inhibition of NO production (IC₅₀ = 37.38 ± 10.27 µg/mL), whereas the ethyl acetate fraction was the strongest suppression of PGE₂ synthesis (IC₅₀ = 86.32 ± 5.51 µg/mL). In contrast, the water fraction did not have a significant effect on these inflammatory markers.

Conclusion: By downregulating NF-κB activity, S. pilifera Benth. effectively modulates the expression of key proinflammatory mediators, including inducible NO synthase (iNOS) and COX-2, ultimately leading to reduced NO and PGE₂ production. These findings suggest that S. pilifera Benth. may exert its anti-inflammatory effects through inhibition of the NF-κB signaling pathway, offering potential therapeutic applications in the management of chronic inflammatory diseases.

Atopic dermatitis (AD) is a chronic, relapsing inflammatory skin disorder with increasing global prevalence. ATOJIN is a natural product composed of Phellodendron amurense Ruprecht (PAR), Schizonepeta tenuifolia (ST), Sophora flavescens (SF), Glycyrrhiza uralensis (GU), and Liriope platyphylla (LP), all known for their anti-inflammatory properties. This study aims to evaluate the therapeutic potential of ATOJIN in a 2,4-dinitrochlorobenzene (DNCB)-induced AD mouse model by assessing its effects on inflammation and immune regulation. The therapeutic efficacy of ATOJIN was evaluated through the analysis of white blood cell subtypes, serum immunoglobulin E (IgE) levels, pro-inflammatory cytokines, and histological assessments of inflammatory and mast cell infiltration, focusing on systemic immune modulation. ATOJIN effectively alleviated AD lesions and symptoms in DNCB-induced mice, demonstrating significant improvements in dermatitis scores, ear thickness, and spleen weight. It also reduced epidermal thickness and the infiltration of inflammatory and mast cells. Furthermore, ATOJIN modulated serum levels of IgE and pro-inflammatory cytokines, including interleukin (IL)-2, IL-4, IL-6, and tumor necrosis factor (TNF)-α, indicating systemic anti-inflammatory effects. These results suggest that ATOJIN mitigates AD symptoms by modulating inflammatory responses, and its efficacy, comparable to or even superior to that of the topical standard-of-care tacrolimus, highlights its potential as a promising standalone oral therapeutic agent for the systemic management of AD.

Micro-fragmented adipose tissue (MFAT) is regarded as one of the simplest and most practical biological preparations for clinical applications in tissue regenerative medicine. The clinical effectiveness of MFAT is attributed to its content of cells and growth factors that facilitate tissue regeneration. In this study, we investigated the biological activity of the secretome derived from cultured MFAT. Our primary focus was on its ability to influence the production of two inflammatory cytokines, RANTES (Regulated and Normal T Cell Expressed and Secreted) and MCP-1 (Monocyte Chemoattractant Protein-1), using ELISA assays, as well as its impact on the expression of Cell Adhesion Molecules (CAMs) on U-937 macrophages via flow cytometry. We also explored the potential of the MFAT secretome to affect the proliferation of both normal and cancer cells. Our results showed that the MFAT secretome inhibited the production of MCP-1 and RANTES, significantly reduced the expression of ICAM-1 (Intercellular Adhesion Molecule 1) on U-937 macrophages, and had no impact on the proliferation of normal or cancer cells. These findings suggest that the MFAT secretome is relatively safe and exhibits anti-inflammatory properties, supporting the idea that its clinical effectiveness in treating joint inflammation may, in part, be due to its paracrine effects.

Background: Mitochondrial-related pathways (MRPs) play a crucial role in cancer metabolism and progression; however, their prognostic value in breast cancer (BC) is still poorly understood.

Methods: We integrated multiomics data to investigate the landscape of MRPs in BC. A mitochondria pathways-associated signature (MPAS) was established using multimachine learning framework and interpreted by SHAP analysis across independent BC cohorts. Additionally, a series of functional experiments were employed to explore the role of RNA exonuclease 2 (REXO2) in BC cells.

Results: MRPs are extensively activated in BC at multiomics level. MPAS demonstrates outstanding predictive performance across multiple BC cohorts, with high scores indicating poor clinical outcomes. Moreover, it was observed that high MPAS scores are closely associated with immunosuppressive states and inflammatory microenvironments. SHAP analysis identified REXO2 as a hub factor of MPAS. Cell-based work confirmed that silencing REXO2 greatly inhibited cell proliferation and induced apoptosis in BC.

Conclusions: Our proposed MPAS could effectively evaluate the prognosis and treatment response of BC patients, providing new reference for clinical decision-making. Furthermore, REXO2 regulates cell proliferation and apoptosis, making it a promising potential therapeutic target for inhibiting BC progression.

Psoriasis, a chronic inflammatory skin disease, arises from a dysregulated interaction between keratinocytes (KCs) and dendritic cells (DCs). We previously identified Rh family C glycoprotein (RHCG) as a key mediator of KC inflammation and DC activation. Here, we demonstrate that luteolin, a bioactive compound derived from the traditional Chinese formula cooling blood and detoxicating formula (CBDF), directly binds to RHCG, as confirmed by multiple computational methods and in vitro experiments. In vitro, luteolin suppressed RHCG expression in KCs, reducing CXCL14 secretion and subsequent DC activation. Spatial transcriptomics (STs) revealed that luteolin preferentially targets DC-enriched spatial domains and restores desmosomal protein expression (e.g., DSC2), which is dysregulated in psoriasis. In vivo, luteolin ameliorated psoriasis-like inflammation in imiquimod-induced mice, lowering Psoriasis Area and Severity Index (PASI) scores and normalizing pathological markers. Our findings indicate that luteolin disrupts KC-DC communication through multiple modes of action, thereby reversing tissue-level pathology and demonstrating its potential as a targeted therapy for psoriasis.

Background: Epithelial-mesenchymal transition (EMT) represents a critical process that facilitates metastatic dissemination and immune evasion in colorectal cancer (CRC); however, the molecular factors that connect EMT to modifications in the immune microenvironment remain poorly elucidated. In this investigation, we identify AJUBA as an essential regulator that mediates the association between EMT and immune modulation in CRC.

Methods: By integrating multi-cohort transcriptomic datasets (The Cancer Genome Atlas (TCGA)-CRC, GSE18105, GSE22598, GSE89076, and GSE110224) with single-cell RNA-seq data (GSE132465), we applied machine learning and deep learning methodologies to comprehensively identify EMT-associated genes demonstrating prognostic significance. AJUBA validation was performed at mRNA and protein levels in a cohort of 90 CRC patient samples using quantitative PCR, Western blotting, and immunohistochemical (IHC) approaches. Functional analyses involved siRNA-mediated knockdown experiments, coupled with evaluations of cell proliferation (CCK-8 assay), migration and invasion (transwell assay), clonogenic capacity (colony formation assay), and in vivo tumor growth in xenograft models. Immune infiltration was assessed via ssGSEA and CIBERSORT algorithms, and spatial transcriptomics data (GSE225857) were used to delineate AJUBA expression within tumor microdomains.

Results: Across multiple CRC cohorts, AJUBA exhibited marked upregulation and showed distinct enrichment in epithelial cells with activated EMT characteristics. Spatial transcriptomic profiling demonstrated AJUBA colocalization with cancer-associated fibroblasts (CAFs) within immune-excluded niches. Enhanced AJUBA expression exhibited a positive correlation with heightened infiltration of M2 macrophages and activation of VEGF/NOTCH signaling cascades. In vivo, AJUBA knockdown led to suppressed tumor growth, reduced Ki-67 proliferation indices, and diminished M2 macrophage abundance. Clinically, elevated AJUBA expression correlated with advanced nodal metastasis and served as an independent predictor of poor overall survival (OS; HR = 4.809, 95% CI: 2.385-9.695, p < 0.001).

Conclusions: AJUBA functions as a key regulator that links EMT to immune modulation, promoting macrophage polarization and facilitating immune evasion in CRC. Through its coupling of EMT activation with proangiogenic signaling, AJUBA represents both a prognostic biomarker and a promising therapeutic target for alleviating immune exclusion in metastatic CRC.

Background: Myasthenia gravis (MG) is an autoimmune disorder in which circular RNAs (circRNAs) are increasingly implicated, with growing evidence supporting their critical role in autoimmune pathogenesis. The role of hsa_circ_0000313 in MG, including its biological functions and mechanisms, remains unknown.

Methods: Agarose gel electrophoresis, RNase R digestion, and Sanger sequencing were employed to verify the circular structure of hsa_circ_0000313, while nucleoplasmic separation experiment was used to determine its subcellular localization. Analysis of hsa_circ_0000313, miR-1224-3p, and MKNK2 expression was conducted via quantitative real-time PCR (qRT-PCR). CCK-8 assay and flow cytometry were employed to evaluate the proliferative capacity and apoptotic rate of Jurkat cells. ELISA detected inflammatory cytokine secretion. Potential interactions involving miR-1224-3p with either hsa_circ_0000313 or MKNK2 were predicted using bioinformatics tools and subsequently validated through dual-luciferase reporter assays. FISH assay was used to detect subcellular colocalization of hsa_circ_0000313 and miR-1224-3p.

Results: We observed that compared to healthy controls, MG patients exhibited increased expression of hsa_circ_0000313 and MKNK2, along with decreased expression of miR-1224-3p. Knockdown of hsa_circ_0000313 and MKNK2, along with overexpression of miR-1224-3p, inhibited the proliferation and secretion of inflammatory factors in Jurkat cells and promoted their apoptosis. Additionally, miR-144-3p was identified as the target miRNA of hsa_circ_0000313, and MKNK2 was identified as the target gene of miR-1244-3p. Hsa_circ_AA0000313 was confirmed to be circular and resistant to RNase R digestion. There was significant colocalization of hsa_circ_0000313 with miR-1224-3p within the cytoplasm. Inhibition of miR-1244-3p reversed the effects of hsa_circ_0000313 knockdown and MKNK2 knockdown on Jurkat cells proliferation, apoptosis, and inflammatory cytokine secretion. In LPS-stimulated Jurkat cells, hsa_circ_0000313 knockdown suppressed the p38 MAPK pathway via the miR-1224-3p/MKNK2 axis, which reduced inflammatory cytokine secretion and cell proliferation as well as promoted apoptosis.

Conclusion: Hsa_circ_0000313 is involved in MG progression by regulating the miR-1224-3p/MKNK2 axis and can act on the p38MAPK pathway to participate in the progression of MG concomitant inflammatory infections, which may provide a promising therapeutic target in MG.

Objectives: Asthma remains a prevalent diagnosis among intensive care unit (ICU) admissions, frequently linked to worsened patient outcomes. Therefore, identifying simple and effective indicators to predict the mortality risk of asthma patients in the ICU is particularly important. Given the unmet need for mortality biomarkers in critical asthma, this study specifically examined neutrophil percentage-to-albumin ratio's (NPAR's) correlation with ICU and in-hospital death.

Materials and methods: We selected 1191 eligible asthma patients from the Medical Information Mart for Intensive Care-IV (MIMIC-IV) 3.0 database for analysis. This study applied a multivariate Cox regression model to explore the relationship between NPAR levels and the risk of mortality in the ICU and during hospitalization. Additionally, we used restricted cubic splines (RCSs) models to investigate the potential nonlinear dose-response relationship between NPAR levels and the risk of mortality in the ICU and during hospitalization.

Results: Among the 1192 enrolled asthma cases, the cohort demonstrated a mean age of 58.1 ± 18.1 years, of which 38.7% were male. Asthma patients with higher NPAR faced substantially greater mortality risks during intensive care and hospitalization (hazard ratio [HR] range 1.75-2.14, p  < 0.01). The relationship between logarithmic transformation (ln) NPAR and ICU and in-hospital mortality rates is nonlinear. Threshold effect analysis showed that when the ln NPAR level exceeded 0.5, the risk of patient mortality significantly increased (HR range 1.574-1.831, p  < 0.05). Additionally, subgroup analysis revealed no significant interactions.

Conclusion: In ICU asthma patients, higher NPAR levels are associated with higher risks of ICU and in-hospital mortality, emphasizing the importance of these findings for early identification and timely intervention in reducing the mortality risk of ICU asthma patients.