Virtual screening identifies specnuezhenide as a potential therapeutic agent for rosacea via KLK5/TLR4/NF-κB pathway inhibition and metabolic modulation
This study demonstrates for the first time that Spe, a novel KLK5 inhibitor, suppresses the inflammatory response mediated by the KLK5/TLR4/NF-κB pathway, accompanied by metabolic remodeling, particularly involving the biosynthesis of unsaturated fatty acids, thereby ameliorating rosacea-like dermatitis.
Abstract
Background Rosacea is a chronic inflammatory skin condition. Excessive LL-37 is produced when serine protease kallikrein-5 (KLK5) is hyperactivated, contributing to the inflammatory response in rosacea. Thus, inhibition of KLK5 has the potential to attenuate inflammation in this condition. Reports suggest that specnuezhenide (Spe), an isomer of oleanolic acid, is effective as an anti-inflammatory agent for the treatment of inflammatory diseases. However, little is known about the role of Spe and its mechanisms in alleviating rosacea. Methods Spe was identified as a potential KLK5 inhibitor through virtual screening and molecular docking, showing high-affinity binding via hydrophobic and hydrogen-bond interactions. Rosacea was induced in mice by intradermal LL-37 injection, followed by intraperitoneal administration of Spe. The severity of skin inflammation was assessed histologically. Serum cytokines, inflammatory gene expression, and protein levels of CD31, MMP-9, CD4, and KLK5 were analyzed by ELISA, RT-qPCR, and immunofluorescence, respectively. Inhibition of the TLR4/NF-κB pathway was examined by Western blot. Untargeted metabolomics was applied to characterize alterations in serum metabolites. Results Spe markedly reduced LL-37-induced skin inflammation, decreased inflammatory cell and mast cell infiltration, decreased MMP-9 expression, and reduced CD31 fluorescence intensity and CD4+ T cell recruitment. The study demonstrated that Spe inhibited the expression of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in serum and suppressed activation of TLR2, MMP-9, KLK5, IL-1β, IL-6, and TNF-α genes. Moreover, Spe inhibited KLK5 expression and blocked activation of the TLR4/NF-κB pathway. Metabolomics identified 43 dysregulated metabolites in rosacea, 26 of which were normalized by Spe treatment. Pathway enrichment analysis suggested that regulation of unsaturated fatty acid biosynthesis is a key mechanism underlying the effects of Spe. Conclusions This study demonstrates for the first time that Spe, a novel KLK5 inhibitor, suppresses the inflammatory response mediated by the KLK5/TLR4/NF-κB pathway, accompanied by metabolic remodeling, particularly involving the biosynthesis of unsaturated fatty acids, thereby ameliorating rosacea-like dermatitis.
Black goji berry (Lycium ruthenicum Murr.) has been recognized for its immune-regulating and anti-cancer potentials, but its key active components and underlying mechanisms remain unclear. Here, we applied LC–MS/MS to characterize the metabolite profile of L. ruthenicum and performed in vitro assays to evaluate its anti-inflammatory and anti-cancer activities. A total of 149 metabolites were identified, with organic acids, carbohydrates, and amino acid derivatives as the dominant components. Isorhamnetin was identified as a representative flavonoid. In LPS-stimulated Calu3 cells, isorhamnetin reduced the secretion and mRNA expression of pro-inflammatory cytokines (IL-6, TNF-alpha, IL-1beta). In cancer cells, isorhamnetin induces ROS accumulation, DNA damage, mitochondrial dysfunction, and caspase-3-dependent apoptosis, with modulation of the AKT signaling pathway. These findings suggest that isorhamnetin may contribute to the anti-inflammatory and anti-cancer effects of L. ruthenicum observed in vitro, providing preliminary experimental evidence for its potential as a functional food ingredient. However, this study is limited by the use of a single cell line (Calu3) and the absence of in vivo validation and pharmacokinetic evaluation; further studies are needed to confirm these observations.
Lei Jiang, Ji-Deng Ma, Zhi-Yuan Li et al.· Frontiers in Nutrition· 0 citations
Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by persistent synovial inflammation and progressive joint destruction. Fibroblast-like synoviocytes (FLS) play a central role in RA pathogenesis by promoting inflammatory signaling and extracellular matrix (ECM) degradation. Although isoliquiritigenin (ISL) exhibits anti-inflammatory and antioxidant activities, its direct molecular target in RA remains unclear. Therefore, this study aimed to identify the molecular target of ISL and elucidate its underlying mechanism. To address this, integrated bioinformatics analysis, target validation (CETSA, DARTS, and molecular docking), together with mechanistic studies in H2O2-treated synovial cells and CIA rats, were performed. Bioinformatics analysis identified NADPH oxidase 4 (NOX4) as one of the overlapping candidate targets of ISL for further investigation. Target engagement of NOX4 by ISL was supported by CETSA, DARTS, and molecular docking analyses. ISL suppressed NOX4-mediated reactive oxygen species (ROS) production, thereby inhibiting inflammasome activation, pro-inflammatory cytokine production, apoptosis, and ECM degradation.ISL improved cell survival and reduced apoptosis in H2O2-treated synovial c ells while preserving ECM integrity. These protective effects were further confirmed in CIA rats, where ISL alleviated arthritis severity, oxidative stress, and cartilage destruction. Collectively, these findings identify NOX4 as a critical upstream regulator linking oxidative stress to inflammasome activation and support pharmacological targeting of the NOX4-ROS-inflammasome axis by ISL as a promising therapeutic approach for RA.
Yu-Chun Chang, W. Kuo, Yen-Pin Chen et al.· Chemico-Biological Interacti...· 0 citations
Endotoxemia is an inflammatory disorder triggered by endotoxins derived from pathogenic bacteria. To identify effective anti-inflammation agents, we screened a chemical compound library and identified Corylifol A as a potent suppressor of LPS-induced inflammation. Corylifol A is a natural flavonoid compound originally isolated from Psoralea corylifolia. Further investigations revealed that Corylifol A dose-dependently inhibited LPS-induced expression of IL-1β, TNF-α, and IL-6 in macrophages, without exhibiting cytotoxicity. Mechanistically, Corylifol A directly bound to IκBα, disrupted IKKβ-IκBα interaction, suppressed IκBα phosphorylation, and subsequently blocked p65 nuclear translocation. In vivo, both prophylactic and therapeutic administration of Corylifol A significantly improved survival rates, reduced inflammatory cytokine levels, and attenuated lung injury in endotoxic mice. Collectively, our findings demonstrate that Corylifol A protects against endotoxemia by targeting IκBα and inhibiting NF-κB activation, highlighting its therapeutic potential for the treatment of endotoxemia and related inflammatory diseases.
Yanxuan Hu, Xiang-Rong Feng, Xibao Zhao et al.· International Immunopharmaco...· 0 citations
Objective Chronic inflammation is a pathological basis for numerous diseases, and soluble epoxide hydrolase (sEH) has emerged as a promising target for anti-inflammatory intervention. This study aimed to identify potential sEH inhibitors derived from the traditional Chinese medicinal herb Oldenlandia diffusa through integrated computational and experimental methodologies. Methods A pharmacophore model was constructed from 41 known sEH inhibitors and utilized to virtually screen 948 metabolites of Oldenlandia diffusa. The identified hits underwent molecular docking, ADMET prediction, induced-fit docking, and 50 ns molecular dynamics simulation. The anti-inflammatory activity of the selected compounds was assessed in LPS-stimulated RAW264.7 macrophages by measuring levels of NO and TNF-α. Results Two compounds, Syringaresinol and 3,7-Di-O-methylquercetin, exhibited favorable ADMET profiles, low cytotoxicity, and stable binding to sEH during molecular dynamics simulation (equilibrated RMSD ∼0.8 nm with fluctuations <0.1 nm; MM-PBSA free energies: –117.49±14.01 and –71.47±13.87 kJ/mol). Both compounds reduced NO and TNF-α levels in a dose-dependent manner in LPS-induced RAW264.7 macrophages. These findings identify metabolites of Oldenlandia diffusa as computationally predicted sEH inhibitors with confirmed anti-inflammatory activity, warranting further direct enzymatic validation. Conclusion Syringaresinol and 3,7-Di-O-methylquercetin from Oldenlandia diffusa are predicted sEH inhibitors with validated anti-inflammatory effects, supporting their further exploration as natural leads for therapeutic applications in inflammation.
Aim Given the limited availability of safe and effective treatments for inflammatory bowel disease (IBD), we applied an integrated network pharmacology approach to systematically map the targets and pathways of cordycepin, a bioactive compound from Cordyceps militaris, in experimental colitis. Methods Cordycepin was administered intraperitoneally during dextran sulfate sodium (DSS) exposure in mice, with efficacy evaluated by the disease activity index (DAI) and histopathological analysis. Network pharmacology analysis (TCMSP, CTD, SEA, BATMAN-TCM, GeneCards, and PharmMapper), molecular docking, and molecular dynamics (MD) simulations were performed to identify and validate potential core targets. AKT1 and tight junction protein ZO-1 expression in colonic tissues was assessed by immunohistochemistry (IHC). The involvement of AKT signaling in cordycepin’s effects on tight junction integrity and mitochondrial function was further investigated in lipopolysaccharide (LPS)-treated Caco-2 cells using the AKT inhibitor MK2206. Results Cordycepin (50 mg/kg) significantly attenuated body weight loss and DAI elevation in DSS-treated mice. A total of 361 putative cordycepin-related targets were identified from six public databases, while 2, 072 UC-related targets were obtained from GeneCards, OMIM, and DisGeNET. A total of 199 overlapping targets were functionally enriched in processes including “TNF signaling pathway”, “PI3K-AKT signaling pathway” and “cellular response to lipopolysaccharide”. The PPI network identified 8 core targets, among which AKT1, NFKB1, RELA and TP53 demonstrated strong binding affinity (binding free energy<-6.0 kcal/mol) with cordycepin in molecular docking and were enriched within the PI3K/AKT pathway. IHC analysis showed that cordycepin reversed alterations of colonic AKT1 and ZO-1 levels in DSS mice. In Caco-2 cells, AKT inhibition with MK2206 attenuated the protective effects on tight junction integrity and mitochondrial function against LPS-induced injury. Conclusion These findings suggest that prophylactic administration of cordycepin, a promising natural compound, alleviates experimental colitis, potentially through modulation of the PI3K/AKT1 signaling pathway and restoration of epithelial barrier integrity.
Wenting Zhang, Minyan Qian, Wenwei Jiang et al.· Journal of Inflammation Rese...· 0 citations
Silicosis is a progressive and irreversible fibrotic lung disease resulting from chronic inhalation of crystalline silica (CS), for which no specific targeted therapy is clinically available. Thus, novel therapeutic agents and their molecular targets are urgently needed. Using GEO-based differential expression analysis and cMap drug prediction, this study identified capsazepine (CPZ) as a potential anti-silicosis agent. In vivo, CPZ dose-dependently ameliorated pulmonary dysfunction, reduced inflammatory cell infiltration and pro-inflammatory cytokine release, and suppressed collagen deposition and epithelial-mesenchymal transition (EMT) in lung tissues of silicosis model mice. In vitro, CPZ directly countered CS-induced inflammatory activation, apoptosis, and oxidative stress in macrophages, while inhibiting aberrant migration and fibrotic activation in fibroblasts, confirming its dual anti-inflammatory and anti-fibrotic effects at the cellular level. Mechanistically, CPZ bound to TRPV1 and subsequently blocked aberrant phosphorylation of the PI3K/AKT signaling pathway, thereby delaying silicosis progression. Collectively, this study is the first to demonstrate that CPZ is a candidate anti-silicosis drug with clinical translational potential, offering a new theoretical and experimental basis for targeted silicosis therapy.
Xiaofei Hu, N. Xu, Chao Liang et al.· European Journal of Pharmaco...· 0 citations
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