Combinatorial chemistry & high throughput screening最新文献

Introduction: This study aims to assess the therapeutic mechanisms of Zishen Yutai Pills (ZYP) in treating recurrent spontaneous abortion (RSA) using network pharmacology analysis and in vitro validation of the molecular pathways relating to angiogenesis.

Methods: An online database screening-based strategy was applied for chemical constituent and target screening-based strategy was applied for chemical constituent and target collection, as well as target prediction. Network pharmacological analysis was conducted using protein-protein interaction data. Molecular docking was further employed to validate the active components of ZYP and the central target proteins. The tube formation assay was conducted in human umbilical vein endothelial cells (HUVECs) with the intervention of active constituents in ZYP. The expression of vascular endothelial growth factor A (VEGFA) and protein kinase B (AKT1) in HUVECs was also quantified by Western Blot (WB).

Results: Network pharmacological analysis revealed 283 target genes by ZYP in the treatment of RSA. Through protein-protein interaction (PPI) and topological analyses, a total of 57 key targets were identified as exerting central roles. Molecular docking proved that quercetin, β-carotene, resveratrol, and atractylenolide I had strong binding to AKT1 and VEGFA. Elevated levels of AKT1 and VEGFA protein expression were observed in WB analysis following intervention with the active ingredient.

Discussion: The findings suggest that ZYP may ameliorate RSA by promoting angiogenesis at the maternal-fetal interface. Key active compounds were predicted to act via the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway, as demonstrated by molecular docking and cell-based assays.

Conclusion: This study illuminates the comprehensive therapeutic effects of ZYP in RSA treatment, providing a foundation for its further exploration.

Introduction: Chan-Hou-Kang-Gao (CHKG) is a classical traditional Chinese medicine (TCM) formula widely used to promote uterine involution and manage postpartum hemorrhage (PPH). However, the pharmacological basis and molecular mechanisms of CHKG in treating PPH remain largely unclear.

Methods: Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was employed to systematically identify the chemical constituents of CHKG. Network pharmacology was used to predict active compounds, potential targets, and relevant signaling pathways. Protein-protein interaction (PPI) analysis and molecular docking were performed to validate the interactions between key compounds and therapeutic targets.

Results: A total of 173 compounds were identified in CHKG, including flavonoids, triterpenoids, and phenolic acids. Integration with public databases revealed 67 common targets shared between CHKG-related compounds and PPH-related genes. Enrichment analysis suggested that CHKG may modulate pathways involved in inflammation, immune regulation, and vascular function. TNF, IL6, and PTGS2 were identified as potential key targets. Molecular docking and molecular dynamics (MD) simulations indicated stable binding of ursolic acid, asiatic acid, and kaempferol to these proteins.

Conclusions: This study provides a systems-level prediction of the mechanisms by which CHKG may exert therapeutic effects in PPH, particularly via inflammation- and coagulationrelated pathways. These findings generate mechanistic hypotheses; however, further in vitro or in vivo experimental validation is required to confirm bioactivity and clinical relevance.

Introduction: The key toxicological constituents and mechanisms of Epimedii Folium and its formulations, such as Xianling Gubao Capsules (XLGB) and Zhuanggu Guanjie Pills (ZGGJ), remain insufficiently understood, particularly when used in combination. The objective of this study is to investigate the hepatotoxic effects and mechanisms of Epimedii Folium and its formulations, XLGB and ZGGJ, using network toxicology, molecular docking, and in vitro validation.

Materials and methods: Potential hepatotoxic components and targets of Epimedii Folium, XLGB, and ZGGJ were screened from multiple databases. PPI networks were constructed, and GO/KEGG enrichment analyses were performed. Molecular docking was used to assess the binding affinities between key components and core targets. In vitro validation was conducted using HepG2 cells to assess cell viability and ROS levels through CCK-8 and HCS assays, respectively.

Results: This study confirms that Sagittatoside A, Epimedin B, and Icariside I are the primary hepatotoxic constituents of Epimedii Folium, capable of targeting core pathways involving KDR, AR, PTGS2, F7, and DPP4. Furthermore, Sagittatoside A and Icariside I significantly elevated ROS levels. The toxic constituents of XLGB and ZGGJ overlapped with those of Epimedii Folium, and Bavachinin and Neobavaisoflavone from PCL were found to exert synergistic hepatotoxic effects. Neobavaisoflavone enhanced the hepatotoxicity of Epimedin B and Icariside I, while Bavachinin showed synergistic toxicity when combined with Sagittatoside A.

Discussion: Molecular docking confirmed strong binding affinities between these compounds and their targets. In vitro experiments demonstrated that Sagittatoside A and Icariside I significantly increased ROS levels. The compound formulations XLGB and ZGGJ shared similar hepatotoxic components and mechanisms. Additionally, Bavachinin and Neobavaisoflavone from PCL synergistically enhanced the hepatotoxicity of Epimedii Folium monomers, providing a modern scientific basis for evaluating compatibility principles in traditional Chinese medicine.

Conclusion: This study comprehensively elucidates the hepatotoxicity and synergistic toxic effects of Epimedii Folium and its formulations XLGB and ZGGJ, offering a modern scientific rationale to guide the safe formulation and compatibility of traditional Chinese medicine.

Introduction: IgA nephropathy (IgAN) is the most common primary glomerular disease. The traditional Chinese formula Mahuang Lianqiao ChixiaoDou Decoction (MLCD) has been demonstrated to alleviate IgAN. However, the potential mechanism remains to be explored. This study was designed to investigate the mechanism of MLCD in improving IgAN through transcriptomics analysis, network pharmacology, and experimental validation.

Methods: The mouse model of IgAN treated with MLCD and a positive control was established. After 8-week treatment, biochemical indicators of renal function and kidney histology were measured. MLCD regulating the potential genes at the transcript level was analyzed. A network pharmacology based on the main components of MLCD identified by HPLC‒MS/MS was adopted. Then, integrated network pharmacology, together with transcriptomics, was performed to predict the potential pathway of MLCD in improving IgAN, which was also validated by ELISA, qRT-PCR, and Western blotting.

Results: MLCD significantly ameliorated kidney damage in IgAN model mice by decreasing the levels of 24-hour proteinuria, serum creatinine, and blood urea nitrogen. The integration of transcriptomics analysis with network pharmacology suggested that the regulation of the MAPK pathway might be a crucial mechanism of action. In vivo experiments confirmed that MLCD attenuated the expression of renal proteins (TLR9, MyD88, IRAK4, p38MAPK, and NF-κB) and reduced the release of inflammatory cytokines (TGF-βand IL-6) in mice.

Discussion: This study clarifies the mechanism of MLCD in treating IgAN; however, further in vitro experiments are needed.

Conclusion: MLCD can reduce the inflammatory response and improve kidney function in IgAN models by regulating the TLR9/MAPK/NF-κB signaling pathway.

Introduction: Huoxue San (HXS) is a traditional Chinese medicinal formulation widely used to treat bone fractures and blood stasis. Comprising seven herbs-Siphonostegia chinensis Benth, Kochia scoparia (L.) Schrad, Scutellaria barbata D.Don, Polygonum cuspidatum Sieb. et Zucc, Arisaema erubescens (Wall.) Schott, Phellodendron chinense Schneid, and Eupolyphaga sinensis Walker-HXS has been administered at Nanjing Chinese Medicine Hospital for over 50 years. It is effective in promoting fracture healing, supporting soft tissue repair, and rarely causing adverse reactions such as skin allergies. The present study aimed to elucidate the molecular mechanisms underlying HXS's therapeutic effects.

Methods: Ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF MS) was used to identify HXS components absorbed into the bloodstream. Network pharmacology, molecular docking, and molecular dynamics simulations were then conducted to explore the active ingredients and their regulatory mechanisms in fracture healing and blood stasis.

Results: Transdermal absorption tests identified 20 active compounds from HXS. Network pharmacology analyses using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform highlighted vanillic acid, demethyleneberberine, palmatine hydrochloride, luteolin, apigenin, and wogonin as key active ingredients. Molecular dynamics simulations further validated the stability, conformational changes, and interactions of these compounds with their target proteins.

Discussion: Analysis of the transdermal absorption samples revealed 291 potential active targets for HXS in treating fractures and blood stasis, of which 159 were common to both conditions. Protein-protein interaction (PPI) network analysis identified core targets including AKT1, ALB, EGFR, STAT3, and CTNNB1. Molecular docking confirmed strong binding interactions between HXS compounds and these core targets, while molecular dynamics simulations validated the stability and mechanistic plausibility of these interactions.

Conclusion: This study provides a systematic elucidation of HXS's molecular mechanisms in fracture healing and blood stasis. Identification of active compounds, core targets, and their interactions offers a scientific basis for the therapeutic effects of HXS and supports the rational development of herbal-medicine-based interventions for fracture management and blood stasis treatment.

Introduction: 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.

Methods: 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.

Results: Polyphenols were extracted by different extraction methods, and they exhibited antioxidant, 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.

Conclusion: 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.

Background: This study aimed to develop and validate a Cuproptosis-Related Gene (CRG) signature for predicting Biochemical Recurrence-Free Survival (BCRFS) and characterizing the Tumor Immune Microenvironment (TIME) in Prostate Cancer (PCa).

Methods: Transcriptomic and clinical data were collected from TCGA (n=405) and GEO (GSE70770, n=203). Consensus clustering based on 10 CRGs defined molecular subtypes. Differentially expressed genes between clusters were subjected to LASSO Cox regression to construct a prognostic signature in the TCGA cohort, followed by validation in GEO and combined cohorts. Quantitative real-time polymerase chain reaction (qRT-PCR) and Immunohistochemistry (IHC) were conducted for experimental validation.

Results: Two CRG-based subtypes were identified, characterized by distinct clinicopathological features, immune checkpoint expression, and BCRFS. A six-gene signature (CALML5, MMP11, UBE2C, ANPEP, TMEM59L, COMP) stratified patients into high- and low-risk groups with significantly different BCRFS (log-rank P<0.001). The model showed good predictive accuracy (AUCs 0.717-0.837 at 1 year, 0.728-0.771 at 3 years, 0.683-0.695 at 5 years) and remained independent of clinicopathological factors. High-risk patients exhibited elevated immune/stromal scores, altered immune infiltration, and higher immune checkpoint expression. qRT-PCR confirmed upregulation of CALML5, MMP11, UBE2C, and COMP in PCa cell lines, while IHC validated differential protein expression of all six genes between PCa and BPH tissues (all P<0.05).

Discussion: This six-gene CRG signature predicts BCRFS and reflects immune heterogeneity in PCa. Its integration into prognostic models may guide personalized management and inform immunotherapy strategies, warranting further validation in prospective clinical studies.

Conclusion: This study initially identified two cuproptosis-related molecules based on the expression patterns of cuproptosis-related genes. In addition, we developed a new cuproptosisrelated molecular signature with great predictive performance for BCRFS and tumor immune environment using six DERRGs (including CALML5, MMP11, UBE2C, ANPEP, TMEM59L, COMP). These findings would be conducive to a deeper cognition of the potential mechanism of cuproptosis of PCa.

Introduction: Icteric hepatitis remains a significant clinical challenge. The integration of Chinese Herbal Medicine (CHM) with conventional Western Medicine (WM) is becoming increasingly common; however, its overall efficacy and safety profile requires systematic evaluation. This study aims to conduct a meta-analysis to assess the safety and effectiveness of using CHM in conjunction with WM to treat icteric hepatitis.

Methods: We conducted a systematic search of PubMed, EMBASE, Cochrane Library, Scopus, Web of Science, and major Chinese databases (CNKI, Wanfang, VIP, CBM) from inception to December 2023 for Randomized Controlled Trials (RCTs) comparing CHM plus WM with WM alone for icteric hepatitis. Two reviewers independently performed study selection, data extraction, and quality assessment using the Cochrane Risk of Bias 2.0 tool. Meta-analysis was performed using RevMan 5.4 software.

Results: Eight RCTs involving 645 participants were included. The combined therapy group demonstrated a significantly higher clinical efficacy rate (RR = 1.22, 95% CI [1.11, 1.34], P < 0.0001) compared to the WM alone group. The combined therapy also resulted in greater improvements in liver function, with significant reductions in ALT (WMD = -58.33 U/L, 95% CI [- 87.75, -28.91]), AST (WMD = -47.11 U/L, 95% CI [-69.83, -24.39]), TBIL (WMD = -48.27 μmol/L, 95% CI [-67.48, -29.06]), and DBIL (WMD = -31.30 μmol/L, 95% CI [-45.16, -17.44]). Furthermore, the integrated approach led to lower levels of inflammatory markers (IL-6, CRP, TNF-α) and faster symptom resolution. There was no significant difference in the incidence of adverse events between the two groups (RR = 0.83, 95% CI [0.44, 1.57], P = 0.56).

Discussion: The pooled evidence suggests that adding CHM to standard WM treatment can enhance therapeutic outcomes by improving liver function and reducing systemic inflammation. The synergistic effects may be attributed to the multi-target pharmacological properties of the herbs used. However, the findings are limited by the high risk of bias and significant heterogeneity across the included studies.

Conclusion: The adjunctive use of CHM with WM appears to be an effective and safe strategy for treating icteric hepatitis. Nonetheless, due to methodological weaknesses in the primary studies, these results should be interpreted cautiously. High-quality, rigorously designed RCTs are needed to confirm these findings.

Introduction: 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.

Methods: 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.

Results: 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/PDL1 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.

Discussion: 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.

Conclusion: 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.

Introduction: Despite the global burden of chronic obstructive pulmonary disease (COPD), its pathogenesis remains elusive, and current therapies fail to halt disease progression. Calycosin, a bioactive isoflavone from Traditional Chinese Medicine (TCM), exhibits antiinflammatory and antioxidant properties, yet its therapeutic potential in COPD remains unexplored.

Methods: We integrated transcriptomic data of human lung tissue from the Gene Expression Omnibus (GEO) database (GSE8581) and human serum metabolomic data from a published research paper to identify COPD-associated pathways. Network pharmacology, including target screening, analysis, and molecule docking, was employed to elucidate the mechanisms of calycosin for COPD therapy.

Results: Multi-omics analysis revealed significant activation of the pyruvate metabolism pathway and glyoxylate/dicarboxylate metabolism pathway in COPD patients, with 590 differentially expressed genes (DEGs) and 116 differentially expressed metabolites (DEMs) identified. Calycosin targets six key regulators (NME1, ALDH2, PGAM1, LDHA, PCNA, RASD1) with binding affinities (-5.1 to -10.2 kcal/mol) validated via molecular docking, implicated in these pathways.

Discussion: This study bridges TCM-derived natural products with modern omics-driven drug discovery, revealing calycosin as a promising COPD intervention by targeting metabolic hubs to mitigate inflammation and metabolic dysfunction, and pioneering an integrated multi-omics and network pharmacology framework for elucidating TCM mechanisms. However, the lack of direct experimental validation in COPD models and the use of data from different biological sources limit the extrapolation of the results. Further in vitro and in vivo experiments are needed.

Conclusions: This integrated analysis highlights distinct metabolic pathway perturbations, specifically in pyruvate and glyoxylate/dicarboxylate metabolism, as key components of COPD pathophysiology and proposes calycosin as a mechanistically grounded candidate for modulating these pathways. This work shifts focus towards metabolic dysregulation as a central therapeutic target, providing a foundation for developing novel strategies to manage COPD beyond symptom control.