Combinatorial Chemistry & High Throughput Screening - Online First

The aim of this study was to investigate the potential mechanisms and therapeutic effects of Total Flavonoids of Drynaria roosii Nakaike (TFRD) on osteoporotic rats following ovariectomy, through modulation of the stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) axis.

Ovariectomized (OVX) osteoporotic rat models were established and treated with TFRD. The effects of TFRD on Bone Mineral Density (BMD), bone microarchitecture, and the expression of genes and proteins related to the SDF-1/CXCR4 axis in rat lumbar vertebrae were assessed using BMD measurement, bone histomorphology analysis, and molecular biology techniques.

In the TFRD treatment group, lumbar spine BMD significantly increased, and trabecular structure improved. Further mechanistic studies revealed that TFRD regulated SDF-1 expression, thereby promoting its binding to the CXCR4 receptor and, in turn, enhancing migration, homing, and osteogenic differentiation of BMSCs. These changes ultimately led to increased bone formation and decreased bone resorption, improving symptoms of osteoporosis.

This study provides novel insights into the molecular mechanism of TFRD gene therapy in OVX osteoporosis rats by elucidating its involvement through the SDF-1/CXCR4 axis and BMSCs-mediated osteogenic differentiation.

These findings serve as a solid experimental and theoretical foundation for developing new anti-osteoporosis drugs. Furthermore, due to its natural plant extract origin, TFRD shows promising clinical application potential and requires further comprehensive investigation.

All-trans retinoic acid (ATRA), a therapeutic mainstay for acute promyelocytic leukemia, is associated with off-target effects on skeletal development, including premature growth plate closure. However, the molecular mechanisms underlying ATRA-induced growth plate senescence remain poorly understood.

Using Sprague-Dawley rats, ADTC5 chondrocyte cell lines, and integrated multi-omics approaches (transcriptome sequencing, weighted gene co-expression network analysis, molecular docking, and functional assays), we investigated how ATRA modulates growth plate development. Animal models were treated with graded ATRA doses, while in vitro studies included cell viability assays, RNA interference, and Western blot analysis to validate interactions in the signaling pathway.

ATRA induced dose-dependent growth plate thinning (high-dose: 59.79 µm vs. control: 511.35 µm) and skeletal growth retardation in rats. Transcriptomic analysis identified ITGB2 as a pivotal gene, with molecular docking revealing a strong binding interaction (-240.25 kcal/mol) between ITGB2 and YAP mediated by hydrogen bonds/salt bridges. Functional experiments revealed that ATRA upregulated ITGB2, which activated YAP, a Hippo pathway effector, thereby suppressing Wnt/β-catenin signaling by inhibiting β-catenin. This led to downregulation of osteogenic markers (Runx2/SOX9) and enhanced growth plate closure. YAP knockdown reversed these effects, restoring β-catenin and downstream target gene expression (c-myc, cyclin D).

Collectively, our findings identify the ITGB2-YAP signaling axis as a novel mechanism underlying ATRA-induced growth plate closure. These findings establish a foundational framework for developing therapeutic strategies, such as targeting ITGB2 or YAP, to potentially delay premature growth plate closure in pediatric patients undergoing ATRA treatment or with related skeletal disorders.

ATRA accelerates growth plate closure through the ITGB2-YAP axis, disrupting Wnt/β-catenin signaling. These findings establish a mechanistic framework for developing therapeutic strategies targeting ITGB2 or YAP to delay premature growth plate senescence in pediatric disorders.

Lingguizhugan Decoction (LGZGD), a traditional Chinese herbal prescription with recognized efficacy in heart failure, has an unclear mechanism against cardiac hypertrophy. This study investigated its protective effects against angiotensin II (Ang II)-induced cardiac hypertrophy and the role of the LITAF signaling pathway.

An in vivo mouse model of cardiac hypertrophy was established via continuous Ang II infusion. LGZGD was administered, and its effects on cardiac function, hypertrophy markers, and pathway proteins were evaluated using echocardiography, histopathology, and molecular techniques. In vitro, H9c2 cardiomyocytes were treated with Ang II to induce hypertrophy; LGZGD-containing serum was applied to assess the impacts on cell size, hypertrophic markers, and signaling pathways. LITAF expression in H9c2 cells was silenced via siRNA to validate its role in LGZGD-mediated anti-hypertrophy.

LGZGD improved cardiac function, reduced cardiomyocyte size, and downregulated hypertrophic markers. It also upregulated LITAF protein expression and suppressed the phosphorylation of ASK1, JNK1/2, and p38 MAPK. LGZGD-containing serum inhibited Ang II-induced H9c2 hypertrophy via activating LITAF and inhibiting the ASK1–JNK/p38 pathway. LITAF silencing reversed these anti-hypertrophic effects, confirming its pivotal role in mediating LGZGD's protective action.

LGZGD alleviates cardiac hypertrophy by activating LITAF and inhibiting the ASK1-JNK/p38 pathway, identifying key therapeutic targets of this formula. These findings advance understanding of LITAF’s non-inflammatory cardiovascular protective roles and provide insights into multi-target strategies for cardiac hypertrophy.

LGZGD attenuates Ang II-induced cardiac hypertrophy by activating the LITAF pathway and inhibiting the ASK1-JNK/p38 signaling cascade.

Polyphenols are important metabolites with polyphenolic structures, which are important bioactive substances distributed in the fruits, roots, bark, leaves, and other tissues and organs of plants. Polyphenols are widely available and have potential research values.

The 157 related references of polyphenols were collected in this review, which were from scientific databases, including PubMed, Web of Science, Elsevier, Willy, SpringerLink, SciFinder, Scopus, ACS publications, Google Scholar, Baidu Scholar, and CNKI.

Polyphenols were extracted by different extraction methods, and they exhibited anti-oxidant, anti-bacterial, anti-cancer, anti-inflammatory, hypoglycemic, and anti-obesity activities. Moreover, polyphenols were widely applied in industry, food, and medicine fields. The biotransformations of anthocyanidins, flavonols, flavanols, flavanones, phenolic acids, tannins, and stilbenes from polyphenols were reviewed in this paper.

The distribution, extraction, bioactivity, biotransformation, and application of polyphenols were comprehensively summarized and analyzed in this review. It will promote the efficient utilization of polyphenols and provide new ideas for future research.

Allergic airway inflammation (AAI), an asthma-like condition, is characterized by Th17/Treg imbalance and PD-1/PD-L1 pathway dysregulation. Yanghe Pingchuan Granules (YP) formulation is clinically used to treat asthma, but its immunomodulatory mechanisms remain unclear.

Using an AAI rat model, the effects of YP were assessed. Flow cytometry was carried out to analyze Th17/Treg proportions. Additionally, the expression levels of Foxp3, ROR?t, IL-10, IL-17, and TGF-1 were measured. PD-L1 siRNA knockdown and overexpression studies were performed to elucidate the role of the pathway.

YP treatment restored the Th17/Treg balance by reducing Th17 and increasing Treg cells. It upregulated IL-10 and TGF-1 while downregulating IL-17. YP inhibited the PD-1/PD-L1 pathway, correlating with improved immune balance and reduced inflammation. PD-L1 modulation confirmed its role in mediating the effects of YP on cellular and cytokine profiles.

The findings indicated that the therapeutic action of YP involves modulation of the Th17/Treg imbalance, likely through inhibition of the PD-1/PD-L1 pathway, thereby shifting thecytokine milieu from a pro- to an anti-inflammatory state.

YP alleviates AAI by modulating the PD-1/PD-L1 pathway to restore Th17/Treg balance and suppress inflammation, thereby revealing its potential immunomodulatory mechanism.

This study aimed to investigate the therapeutic effect and mechanism of Yangweishu granules (YWS) for stress gastric ulcer (SGU).

The rat SGU model was established using the water immersion restraint stress method (WIRS). The therapeutic effect of YWS was evaluated by observing the histological changes of the stomach tissue, the levels of inflammatory factors, and oxidative stress. Meanwhile, the potential core targets and signaling pathways of YWS in anti-SGU action were analyzed using network pharmacology methods, and the related pathways were experimentally verified.

YWS decreased the expressions of TNF-α, IL-1β, IL-6, and MDA in serum, and increased the levels of IL-4, IL-10, SOD, and GSH-PX. Network pharmacology analysis suggested that YWS may act on the targets of TLR4, AKT1, IL-10, TNF-α, IL-1β, and TP53 through the toll-like receptor pathway to treat SGU. RT-PCR, immunohistochemical, and Western blot results showed that YWS significantly inhibited the TLR4/MyD88/IKB-α pathway. Molecular docking results showed that the main active component of YWS could bind tightly to the TLR4 protein.

This study established an animal model of SGU and preliminarily investigated the therapeutic effects and mechanism of YWS. To more comprehensively evaluate its application value in the treatment of peptic ulcers, subsequent studies should construct various types of ulcer models, further systematically assess the efficacy of YWS, and deeply explore its potential mechanism.

YWS could alleviate WIRS-induced SGU in rats, and its potential mechanism was found to involve the inhibition of the TLR4/MyD88/ IKB-α signaling pathway.

Gumibao decoction, a traditional Chinese herbal prescription, has demonstrated promising effects in treating osteoporosis; however, the underlying mechanism remains unclear. We investigated the regulatory mechanisms of Gumibao decoction in osteoporosis through network pharmacology analysis and validated its therapeutic effects using animal experiments.

The TCMSP database was used to screen the bioactive constituents of Gumibao decoction and identify their associated targets. Disease targets for osteoporosis were acquired through the GeneCards, PharmGKB, DrugBank, OMIM, and TTD databases. A Protein–Protein Interaction (PPI) network was generated. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were subsequently performed for the core targets identified in the PPI network. Moreover, a glucocorticoid-induced osteoporosis model was established in Sprague–Dawley (SD) rats by administering methylprednisolone and dexamethasone, and the regulatory effects of Gumibao decoction on osteoporosis were evaluated using bone mass detection and western blotting.

The network pharmacology analysis identified quercetin, (−)-Epigallocatechin-3-Gallate (EGCG), kaempferol, luteolin, and fisetin as the primary bioactive components of Gumibao decoction. Core target proteins included AKT1, TP53, JUN, CTNNB1, and IL1B. The KEGG pathway enrichment analysis revealed the significant involvement of the TGF-β signalling pathway, osteoclast differentiation, and the MAPK signalling pathway in mediating its anti-osteoporotic effects. In vivo validation demonstrated that Gumibao decoction significantly ameliorated glucocorticoid-induced reductions in Bone Mineral Density (BMD) and deterioration of bone microstructure. Furthermore, protein expression analysis revealed significantly reduced levels of Smad4, phospho-Smad2/3, and TGF-β1 in the model group compared with the blank control group. Notably, all Gumibao decoction treatment groups exhibited significant upregulation of Smad4, P-Smad2/3, and TGF-β1 expression compared with the model group, validating the network pharmacology predictions implicating the TGF-β pathway.

Research on the components of Gumibao decoction has shown that it can regulate homeostasis between osteoblasts and osteoclasts through multiple targeted pathways, thereby positively modulating bone density, bone microstructure, and bone markers. This ultimately inhibits osteoclast differentiation and stimulates osteoblast proliferation, effectively alleviating and preventing osteoporosis.

Gumibao decoction effectively improves glucocorticoid-induced osteoporosis, potentially by upregulating the expression of TGF-β1, P-Smad2/3, and Smad4 through the TGF-Β/Smad pathway, promoting bone formation, and restoring bone metabolic balance in glucocorticoid-induced osteoporosis rats.

Allergic rhinitis (AR) has long been considered to be associated with chronic adenotonsillar disease (CATD). However, their causal relationship remains unclear. This study aims to investigate the causal relationship between AR and CATD and to examine the mediating role of chronic rhinosinusitis (CRS) in this association.

This study employed a two-sample Mendelian randomization (MR) design using genetic instrumental variable analysis. Data for allergic rhinitis (AR) were obtained from the MRC IEU OpenGWAS data infrastructure, data for chronic adenotonsillar disease (CATD) from the FinnGen biobank, and data for chronic rhinosinusitis (CRS) from the GWAS Catalog. Several MR methods were applied. In addition, a two-step MR approach was used to investigate the mediating role of CRS in the relationship between AR and CATD.

MR analysis identified a positive correlation between AR and CATD. IVW and weighted median analyses showed significant causal effects (beta = 0.55, 95% CI: 0.26 to 0.84); p <0.001). No causal association was found between CATD and AR. AR and CRS showed a positive correlation (beta = 1.38, 95% CI: 0.78 to 1.98; p = 6.5 × 10^-6). CRS had a beta value of 0.15 (95% CI: 0.06 to 0.24; p = 0.001) for CATD. CRS mediates 37.6% of the AR to CATD pathway (mediation effect = 0.20, 95% CI: 0.04 to 0.37; p = 0.013).

These findings indicate that AR may contribute to CATD risk through CRS, highlighting the need for further research to explore underlying biological mechanisms and validate these findings.

This study suggests a positive causal relationship between AR and CATD, with CRS acting as a mediator.

Polygonatum sibiricum (P. sibiricum) possesses antioxidant and anti-inflammatory activities. We explored the multi-target mechanisms of P. sibiricum against hepatocellular carcinoma (HCC), aiming to improve its poor prognosis.

Active compounds and disease targets of P. sibiricum were retrieved from the TCMSP and CTD databases. A PROTEIN-PROTEIN INTERACTION (PPI) network was constructed using the STRING database, and functional enrichment was performed with the clusterProfiler package. A compound-target-pathway network was developed in Cytoscape. Immune infiltration was assessed via CIBERSORT and ESTIMATE algorithms, while ligand-target binding was evaluated by molecular docking and 100-ns molecular dynamics (MD) simulations. In vitro experiments were performed to explore the expression and functions of the key genes.

We screened 9 active components, 87 putative targets, and 240 HCC-related genes.

20 overlapping targets were used to construct a PPI network. Network analysis identified baicalein and 4 core targets (MMP9, AKT1, TP53, and PTGS2). Molecular docking and 100-ns MD simulations confirmed stable ligand-protein binding. Immune profiling showed that higher expression of the core targets was related to higher StromalScore, ImmuneScore, and lower tumor purity. Enrichment analysis revealed that these genes were involved in critical pathways, including angiogenesis, EMT, and inflammation response. Functionally, MMP9 knockdown suppressed HCC cell proliferation, migration, and invasion.

P. sibiricum, particularly through baicalein targeting FOS/MMP9/AKT1/ TP53/PTGS2, inhibited HCC development by modulating EMT/angiogenesis pathways and immune milieu. However, these findings required further verification.

Baicalein was identified as an active compound targeting 5 crucial genes to suppress HCC progression, uncovering a new anti-HCC mechanism of P. sibiricum.

Cartilage defects (CDs) are orthopedic conditions with limited regenerative potential. This study aimed to identify endoplasmic reticulum (ER) stress-related biomarkers and construct a diagnostic model to enhance the early detection of CD.

This study analyzed the transcriptomic dataset GSE129147 to identify ER stress-related differentially expressed genes (ERSRDEGs) between CD and control tissues using the limma package (version 3.58.1). Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses were employed for functional enrichment. Immune infiltration was assessed using cell-type identification, which involved estimating the relative subsets of RNA transcripts and single-sample gene set enrichment analysis. Diagnostic models were constructed using logistic regression, support vector machine, and least absolute shrinkage and selection operator regression.

Twenty ERSRDEGs were identified, with CYBB, ATP6V1A, and TNFRSF12A significantly upregulated in CD samples. GO and KEGG analyses highlighted oxidative stress response and extracellular matrix remodeling as key mechanisms in CD pathogenesis. Immune analysis revealed an increase in regulatory T cells and a reduction in CD8⁺ T cells. TNFRSF12A showed strong immune associations and, together with TWIST1 and ATP6V1A, formed the final preliminary diagnostic model. The preliminary LASSO model achieved satisfactory predictive accuracy (AUC: 0.7–0.9).

These findings suggest that ER stress and immune imbalance jointly contribute to cartilage degeneration. The identified genes, particularly TNFRSF12A, TWIST1, and ATP6V1A, not only serve as potential biomarkers but also provide preliminary evidence for new mechanistic insights into stress–immune crosstalk in CD.

This study reveals the key roles of ER stress and immune dysregulation in CDs. Moreover, the ERSRDEG-based diagnostic model provides preliminary bioinformatics evidence and potential molecular indicators for targeted diagnostics and therapies.