Multi-target Mechanism Research Articles

Gastric cancer (GC), a life-threatening malignancy with profound global health impacts, remains a cardinal focus of biomedical research. Recently, astragaloside IV (AS-IV), a bioactive triterpenoid saponin derived from Astragalus mongholicus Bunge, has garnered substantial attention for its multifaceted anticancer properties in preclinical investigations. This review systematically synthesizes current evidence on the molecular mechanisms underlying AS-IV's inhibitory effects against GC, encompassing programmed cell death pathways (apoptosis, autophagy, pyroptosis, ferroptosis), tumor angiogenesis, tumor microenvironment modulation, Helicobacter pylori and inflammatory signaling networks. Many studies demonstrate that AS-IV can inhibit the development of GC through multi-target and multi-pathway mechanisms, making it a well-deserved nemesis of GC. Notably, although AS-IV has emerged as a potential candidate for GC therapy, it suffers from problems such as single research model, unclear toxic and side effects, and poor bioavailability. These seriously hinder the efficiency of AS-IV in the treatment of GC. In the future, we can design and implement a series of in vivo and in vitro experiments to further explore and clarify the mechanism of action of AS-IV in the treatment of GC. It is encouraged to carry out a number of high-quality clinical controlled studies to further prove the effectiveness and safety of AS-IV. In addition, we can also use emerging technologies (such as nanotechnology) to improve the bioavailability of AS-IV, bringing more hope to GC patients.

She Medicine, a traditional therapeutic system from China's She ethnic group, shows promise in cancer treatment. This paper provides a comprehensive review of She medicinal herbs, focusing on their anticancer activities and underlying mechanisms. Compared to widely studied traditional medicines (e.g., Traditional Chinese Medicine), She Medicine exhibits unique ethnopharmacological traits, such as localized plant usage and multi-target mechanisms involving apoptosis induction, immune modulation, and tumor microenvironment regulation. Key herbs like Pimpinella diversifolia and Melastoma dodecandrum showing significant anticancer potential due to their bioactive compounds such as flavonoids, quercetin, and gallic acid. For example, homoharringtonine (HT), a She-derived alkaloid, targets Smad3/TGF-β pathways in non-small cell lung cancer and synergizes with chemotherapy in leukemia treatment, as evidenced by preliminary clinical trials. However, challenges persist, including resource shortages, insufficient mechanistic studies, and a lack of quality control standards. Future research should integrate multi-omics and bioengineering approaches to standardize She Medicine and bridge its traditional use with modern therapies such as immune checkpoint inhibitors. Overall, She medicinal herbs hold great promise for cancer treatment and warrant further exploration to unlock their full potential in modern medicine.

Cancer remains one of the most formidable challenges to human health; hence, developing effective treatments is critical for saving lives. An important strategy involves reactivating tumor suppressor genes, particularly p53, by targeting their negative regulator MDM2, which is essential in promoting cell cycle arrest and apoptosis. Leveraging a drug repurposing approach, we screened over 24,000 clinically tested molecules to identify new MDM2 inhibitors. A key innovation of this work is the development and application of a selective cleaning algorithm that systematically filters assay data to mitigate noise and inconsistencies inherent in large-scale bioactivity datasets. This approach significantly improved the predictive accuracy of our machine learning model for pIC50 values, reducing RMSE by 21.6% and achieving state-of-the-art performance (R2 = 0.87)—a substantial improvement over standard data preprocessing pipelines. The optimized model was integrated with structure-based virtual screening via molecular docking to prioritize repurposing candidate compounds. We identified two clinical CB1 antagonists, MePPEP and otenabant, and the statin drug atorvastatin as promising repurposing candidates based on their high predicted potency and binding affinity toward MDM2. Interactions with the related proteins MDM4 and BCL2 suggest these compounds may enhance p53 restoration through multi-target mechanisms. Quantum mechanical (ONIOM) optimizations and molecular dynamics simulations confirmed the stability and favorable interaction profiles of the selected protein–ligand complexes, resembling that of navtemadlin, a known MDM2 inhibitor. This multiscale, accuracy-boosted workflow introduces a novel data-curation strategy that substantially enhances AI model performance and enables efficient drug repurposing against challenging cancer targets.

Preeclampsia is a life-threatening pregnancy disorder characterized by hypertension and multiorgan dysfunction, posing significant risks to both maternal and fetal health. Although low-dose aspirin is widely recommended for preventing preeclampsia, the underlying mechanisms of action are still poorly understood, which hinders the optimization of therapeutic strategies. We developed an in vitro hypoxia-induced preeclampsia model using human trophoblast organoids to replicate key pathological features. RNA sequencing identified dysregulated pathways and molecular targets. Functional assays assessed the effects of aspirin on trophoblast proliferation, mitochondrial activity, and hormonal regulation, focusing on the PI3K (phosphatidylinositol 3-kinase)-AKT (protein kinase B) pathway and CYP (cytochrome P450) enzymes. We also analyzed the effects of aspirin in the N'-nitro-L-arginine-methyl ester hydrochloride rat model. The hypoxia-induced preeclampsia model successfully mimicked clinical hallmarks, including elevated sFLT-1 (soluble fms-like tyrosine kinase 1)/PlGF (placental growth factor) ratios and oxidative damage. RNA sequencing revealed significant suppression of the PI3K-AKT-mTOR (mammalian target of rapamycin) pathway and dysregulation of CYP enzymes. Aspirin treatment restored the sFLT-1/PlGF balance, reactivated the PI3K-AKT-mTOR pathway, and improved mitochondrial function, enhancing trophoblast proliferation. Furthermore, aspirin regulated CYP by increasing CYP19A1 (cytochrome P450 1A1) and inhibiting CYP1A1 (cytochrome P450 1A1), thereby improving placental hormonal homeostasis. This study clarifies aspirin's multitarget mechanisms in alleviating preeclampsia, which include restoring the sFLT-1/PlGF balance, activating the PI3K-AKT-mTOR signaling pathway, optimizing mitochondrial function, and regulating CYP-mediated hormonal metabolism. These findings provide a mechanistic basis for aspirin's clinical effectiveness in preventing preeclampsia.

Accumulating clinical evidence demonstrates the therapeutic potential of Traditional Chinese Medicine (TCM) in mitigating depressive disorders. This research focuses on quercetin, a principal bioactive constituent shared among five classical TCM antidepressant formulations, to systematically decode its multi-target mechanisms via an integrative framework combining neuroinflammatory modulation and synaptic plasticity regulation. A tripartite experimental design was implemented. Firstly, bibliometric analysis systematically screened antidepressant TCM prescriptions and their bioactive components. Secondly, network pharmacology delineated the therapeutic mechanisms of quercetin - a key phytochemical identified through prior analysis. Finally, we established a chronic unpredictable mild stress (CUMS)-induced depression-like behavior model in mice for validation. Bibliometric analysis showed that the clinical efficacy of 5 TCM antidepressant prescriptions were identified by evidence-based medicine. In these prescriptions, Radix Bupleuri, Rhizoma Cyperi, and Radix Glycyrrhizae were the most commonly used herbs, while Quercetin was identified as the shared bioactive nexus across these prescriptions. Network pharmacology analysis revealed that quercetin may be closely related to PI3K/AKT pathway in depression. And results of animal experimentation showed that quercetin could improve depression-like behaviors and restore neurotransmitters levels. Concurrently, quercetin may inhibit neuroinflammation and ameliorate synaptic ultrastructural by PI3K/AKT pathway. The present study elucidated the mechanism of quercetin, an active ingredient in TCM prescriptions, in the treatment of depression through data mining, network pharmacology prediction, and experimental validation. This integrated research method will provide a new perspective for the development of TCM.

IntroductionPsoriasis is a chronic immune-mediated inflammatory skin disease. Xiaoyin Jiedu Decoction (XYJDY) is a traditional Chinese medicinal formula that has demonstrated significant clinical efficacy in alleviating psoriatic symptoms; however, its underlying pharmacological mechanisms remain unclear.MethodsWe employed network pharmacology, machine learning–based target screening, and functional enrichment to identify key targets and pathways. Single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics (ST) were used to validate gene expression. An IL-17A–induced HaCaT cell model was established for in vitro validation.ResultsAKR1B10 was identified as the core therapeutic target of XYJDY in psoriasis. It was markedly upregulated in psoriatic skin lesions, primarily enriched in keratinocytes, and its expression demonstrated positive correlations with multiple pro-inflammatory immune cell subsets. In vitro experiments showed that XYJDY-medicated serum significantly downregulated AKR1B10 expression in IL–17A–stimulated HaCaT cells.ConclusionThis study reveals that the multi-component formula XYJDY exerts anti-psoriatic effects through a multi-target synergistic mechanism, in which AKR1B10 is a potential core target. These findings provide a theoretical foundation for further exploration of the molecular mechanisms underlying the efficacy of XYJDY in psoriasis treatment.

Autophagy induced by itraconazole and ritonavir was found involved in the pathogenesis of C. albicans. This study was designed to explore the possible molecular mechanism of itraconazole and ritonavir in the treatment of Candida albicans infection through autophagy pathway. The overlapping targets of itraconazole and ritonavir, and those-related to C. albicans and autophagy were screened. Then the core targets were identified by protein-protein interaction (PPI) network analysis. Gene enrichment analysis of targets and the drug-target-pathway-disease network was constructed. The interactions between itraconazole, ritonavir and core targets were analyzed by molecular docking and molecular dynamics simulation. Finally, the core target-miRNA interaction network was constructed to predict candidate miRNAs. PPI network showed that PIK3R1, RELA, STAT3, HSP90AA1, TP53, JUN, GRB2, EGFR, ESR1 and TNF were potential core targets of autophagy therapy for C. albicans infection with itraconazole and ritonavir. Enrichment analysis showed that the two drugs may regulate the autophagy process through pathways including PI3K-AKT, IL-17, MAPK, Toll-like receptor, JAK-STAT and NF-κB. Molecular docking analysis indicated that itraconazole and ritonavir possess strong binding affinities with the cote target proteins, with binding free energies ranging from -5.6 to -9.5kcal/mol. Key interactions were identified at the active sites of the targets, suggesting stable ligand-receptor complex formation. Itraconazole docked to PIK3R1 through SER-78 and GLU-82 (-9.3kcal/mol), and ritonavir docked to PIK3R1 through ASN-85, GLU-1011 and arginine (ARG)-1088 (-7.7kcal/mol). Molecular dynamics simulation of itraconazole and ritonavir with representative target genes lasted for 100ns showed the structures of the formed complexes remained stable throughout. Finally, the candidate miRNAs including miR-486-5p, miR-411-5p.1 and miR-296-5p were identified. Network pharmacological analysis showed a multi-target and multi-pathway molecular mechanism of itraconazole and ritonavir in the treatment of C. albicans infection, and provided a theoretical basis for subsequent studies.

BackgroundClostridium perfringens is a pathogen that secretes multiple toxins, impacting humans and animals. It can cause intestinal diseases such as necrotic enteritis. Although tannins inhibit C. perfringens proliferation, the precise underlying mechanisms are unclear.ObjectiveThis study integrated transcriptomics and metabolomics to systematically investigate the mechanism by which tannins, specifically pentagalloylglucose (PGG) and tannic acid (TA), inhibit C. perfringens and potential pathways to alleviate infection in vivo.ResultsIon concentration measurements, flow cytometric analysis, and transmission electron microscopy revealed that PGG and TA damaged the cell membrane structure of C. perfringens, triggering cytoplasmic content leakage. Additionally, PGG and TA significantly affected C. perfringens at the transcriptional and metabolic levels. Bioinformatics analysis revealed that PGG and TA induced amino acid restriction, disrupted energy metabolism, and impeded the ability of C. perfringens to sense and respond to the external environment. In an in vitro C. perfringens-infected intestinal cell model, PGG and TA bound α toxin, significantly reduced the mRNA expression of inflammatory factors, and improved intestinal barrier function and cell viability. Compared to PGG, TA exhibited stronger inhibitory activity against C. perfringens and binding to α toxin. In vivo, PGG and TA alleviated C. perfringens-induced weight loss in mice, improved intestinal villi morphology, and reduced intestinal inflammation and tight junction gene dysregulation.ConclusionThese findings indicate that tannins inhibit C. perfringens, improve gut tissue integrity and reduce inflammation, demonstrating their multi-target effects of resisting intestinal diseases caused by harmful bacteria. This offers new insights for plant polyphenol-based strategies against necrotic enteritis.Graphical

BackgroundPercutaneous coronary intervention (PCI) is a cornerstone treatment for coronary artery disease (CAD), yet opportunities remain to improve clinical outcomes, symptom management, and long-term prognosis. Traditional Chinese Medicine (TCM), with its multi-target and multi-pathway mechanisms, offers a promising complementary approach to enhance PCI efficacy.MethodsA systematic search was conducted in PubMed and Web of Science using the terms: (“Percutaneous Coronary Intervention” AND “Traditional Chinese Medicine”) and (“Percutaneous Coronary Intervention” AND “Chinese Herbal Drugs”). Randomized controlled trials (RCTs) with ≥100 participants were included to evaluate TCM’s clinical efficacy in PCI. Pharmacological studies were also reviewed to explore underlying mechanisms.ResultsA review of 20 RCTs showed that TCM plays multiple roles in CAD treatment during PCI. Specific interventions such as Danhong Injection, Tongxinluo Capsule, and Shenzhu Guanxin Granule were found to alleviate angina symptoms, restore cardiac function, reduce cardiac biomarkers, prevent no-reflow/slow-flow phenomena, inhibit in-stent restenosis, and improve prognosis while reducing complications. Mechanistically, TCM exerts its effects through antiplatelet action, anti-inflammation, inhibition of smooth muscle proliferation, vasodilation, microcirculation improvement, and endothelial protection.ConclusionThis systematic review highlights the complementary benefits of TCM in PCI for CAD patients. Effective interventions such as Danhong Injection and Tongxinluo Capsule contribute to symptom relief, cardiac function restoration, restenosis inhibition, and prognosis improvement. These benefits are linked to TCM’s multi-target mechanisms, including anti-inflammatory and antiplatelet effects. Future high-quality studies are needed to further validate these findings and refine clinical applications.

Atopic dermatitis (AD) is a chronic inflammatory skin disorder with limited effective treatment. Sophora flavescens Aiton (Kushen) exhibits anti-inflammatory properties, but its key active components and mechanisms against AD remain unclear. This study aimed to identify its therapeutic compounds and the underlying molecular mechanisms. Network pharmacology and ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry were applied to screen for the potential bioactive compounds from Kushen, which were validated in AD mouse models. Formononetin (FMN) was topically administered to evaluate its curative efficacy through histopathological and immunofluorescent staining of skin lesions. Transcriptomic profiling explored the molecular mechanisms, with subsequent validation of neutrophil extracellular traps (NETs)-associated markers by immunofluorescent staining and western blotting. Co-treatment with PAD4 inhibitor GSK484 and molecular docking were used to study the underlying mechanisms. The study identified FMN as an important active component in Kushen, which showed therapeutic effects comparable to dexamethasone in AD models. FMN normalized cutaneous hyperplasia, reduced infiltration of CD3+ T cells, mast cells, and neutrophils, and suppressed inflammatory cytokine expression. Mechanistically, FMN inhibited NETs formation evidenced by decreased levels of citrullinated histone H3 (citH3), myeloperoxidase (MPO), and peptidylarginine deiminase 4 (PAD4). GSK484 co-treatment showed non-synergistic effects, suggesting that FMN suppresses NETs formation primarily through PAD4/MPO inhibition, which was confirmed by molecular docking. These findings highlight FMN as a primary anti-AD constituent of Kushen, therapeutically suppressing NETs-driven crosstalk of innate and adaptive immunity. FMN's multitarget mechanisms provide mechanistic insights and position it as a candidate for AD treatment.

Exploring chemical constituents and anti-inflammatory mechanisms of Semiaquilegiae Radix via an integrated strategy combining UHPLC-Q-TOF–MS analysis, network pharmacology and molecular docking

Gliotoxin (GT) is a sulfur-containing secondary metabolite that belongs to a class of naturally occurring 2,5-diketopiperazines produced by fungi. Although GT production has been observed only in a few species, GT is the most studied fungal secondary metabolite, and the GT biosynthetic gene cluster (BGC) is broadly present in filamentous fungi. GT has a multitarget mechanism of action: It is fungicidal and bacteriostatic, it induces apoptosis in mammalian cells, and it modulates phagocytosis and neutrophil attraction. GT is important for Aspergillus fumigatus virulence and pathogenesis in humans and in animals and for Trichoderma spp. symbiotic and antagonistic behavior. GT is also toxic for producer and nonproducer organisms. Consequently, very sophisticated mechanisms of GT self-protection have evolved in producers; some of these protective mechanisms are also found in nonproducer organisms. This review discusses the distribution of the GT BGC among filamentous fungi and discusses GT biosynthesis, mechanisms of action and self-defense, and ecological properties.

Objective: Cancer remains a global health challenge due to the limitations of conventional therapies, including drug toxicity and resistance. This study aims to explore the anticancer potential of Rubiaceous plant species by investigating their bioactive phytochemicals, molecular targets, and pharmacological pathways, with a particular focus on breast cancer. Materials and Methods: A network pharmacology approach was employed to identify therapeutic compounds and their molecular targets. Disease-related targets were sourced from GeneCards and the Therapeutic Target Database (TTD). Cytoscape and STRING were used to construct interaction networks. Gene Ontology (GO) and KEGG pathway enrichment analyses were performed to elucidate biological functions and pathways. Molecular docking studies were conducted to assess the binding affinities of key phytoconstituents. Results: A total of 1,435 biological processes and 173 pathways were associated with breast cancer. Molecular docking revealed Quercetin as the most potent compound with a binding affinity of -34.92 kcal/mol. Other compounds such as Acacetin, Resveratrol, and Apigenin exhibited lower, but significant, binding affinities. Rubiaceous plants, including Alibertia myrciifolia, Anthocephalus cadamba, and Camptotheca acuminata, were identified to contain flavonoids, alkaloids, and anthraquinones with demonstrated anticancer effects, including apoptosis induction and DNA damage. Conclusion: Rubiaceous plants exhibit promising anticancer potential through multi-target mechanisms. Regulatory oversight is crucial to ensure the safety and efficacy of these herbal therapies. Further research is warranted to isolate active compounds, understand their molecular mechanisms, and validate their clinical relevance for integration into modern oncology.

ABSTRACTProstate cancer (PCa) is the second leading cause of cancer‐related death among American men, and its long latency offers a window for chemopreventive strategies. Phytochemicals, with their diverse impacts on cancer cell growth and metabolism, represent promising candidates for such strategies. Combining compounds like curcumin (Curc) and ursolic acid (UA), which target multiple pathways, can be advantageous in slowing tumor progression. Previous studies revealed the synergistic effects of Curc + UA in reducing tumor growth in a PCa allograft model. In this study, diet‐based interventions were evaluated using two transgenic mouse models of PCa. Mice fed a Curc + UA‐enriched diet exhibited significant inhibition of prostate tumor progression compared to single‐agent diets in both HiMyc and PTEN knockout mouse models. Protein analyses of ventral prostate tissues from HiMyc mice indicated that the combination suppressed oncogenic signaling pathways, including STAT3, AKT, and mTORC1, while modulating cell regulatory proteins to inhibit tumor cell proliferation. Furthert mechanistic studies in mouse and human PCa cell lines confirmed that Curc + UA exerted pleiotropic effects by influencing oncogenic signaling, cell cycle regulation, mitochondrial function, unfolded protein response (UPR), and apoptosis, collectively contributing to its synergistic efficacy. These findings highlight the potential of Curc + UA to inhibit PCa progression through multitargeted mechanisms. The combination's superior efficacy over single agents underscores its promise as a chemopreventive or therapeutic strategy. This study provides a strong rationale for further mechanistic investigations and clinical development of Curc + UA for PCa prevention and treatment.

The Oropouche virus (OROV), an emerging arbovirus transmitted by arthropods, has caused significant outbreaks in South and Central America, with over half a million reported cases. Despite its public health threat, no approved vaccines or antiviral treatments exist for Oropouche fever (OF). This study explores the potential of epigallocatechin-3-gallate (EGCG), a bioactive polyphenol from green tea, as an antiviral agent against OROV using computational approaches. Due to the lack of experimentally resolved OROV protein structures, we employed AlphaFold2 to predict 3D models of key viral proteins, including RNA-dependent RNA polymerase (RdRp), envelopment polyprotein, nucleoprotein, and glycoprotein Gc. Molecular docking revealed strong binding affinities between EGCG and these targets, with particularly high interactions for RNA polymerase (-7.1kcal/mol) and envelopment polyprotein (-8.7kcal/mol), suggesting the inhibition of viral replication and entry. Protein-protein interaction (PPI) network analysis identified critical human host genes (e.g., FCGR3A, IRF7, and IFNAR1) involved in immune responses, while Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses highlighted enriched antiviral and inflammatory pathways. ADMET profiling indicated challenges in EGCG's bioavailability, including poor gastrointestinal absorption and blood-brain barrier permeability, but its low toxicity and natural origin support its potential as a lead compound. These findings suggest that EGCG may disrupt OROV infection through multi-target mechanisms, warranting further experimental validation. This study provides a foundation for developing EGCG-based therapeutics against OROV and underscores the utility of computational methods in antiviral drug discovery.

Diabetes mellitus, one of the major causes of death globally, is a serious health concern since it is characterised by impaired insulin function or production. Concerns about the negative consequences of traditional drugs have drawn attention to herbal remedies. The purpose of this article is to review the efficacy of plant-based remedies (herbal medications) in the management of diabetes, as documented in the various literature. This review highlights that herbal medicines offer a promising complementary approach to diabetes management, due to their affordability and multi-target mechanisms. Most evaluated plants demonstrated significant anti-diabetic effects, including improved glycaemic control, insulin sensitivity, and metabolic regulation. Notably, apple cider vinegar was most effective in reducing fasting blood glucose and HbA1C. Herbal formulations, rich in flavonoids, tannins, phenolics, and alkaloids, support pancreatic function and glucose regulation. While tea extracts showed no benefit, Aloe vera, Fenugreek, Psyllium fibre, and Nigella sativa emerged as effective adjuncts. Rigorous clinical validation remains essential for broader therapeutic acceptance.

Natural compounds remain a valuable source of anticancer agents due to their structural diversity and multi-targeted mechanisms of action. Roburic acid (RA), a tetracyclic triterpenoid, has been identified as a substance capable of inhibiting key NF-κB and MAPK signaling pathways through direct interaction with TNF-α, as well as preventing the production of inflammatory mediators and cancer progression. In this study, we evaluated the biological activity of RA against a panel of human cancer cell lines—DLD-1, HT-29, and HCT-116 (colorectal), PC-3 (prostate), and BxPC-3 (pancreatic)—as well as two non-malignant lines: WI-38 (fibroblasts) and CCD-841 CoN (colon epithelium). RA exhibited a concentration-dependent inhibitory effect on cancer cell metabolic activity, with colorectal cancer cells showing relatively higher sensitivity, particularly at shorter incubation times. To distinguish between cytotoxic and cytostatic effects, we performed trypan blue exclusion combined with a cell density assessment, clonogenic assay, and BrdU incorporation assay. The results from these complementary assays confirmed that RA acts primarily through an antiproliferative mechanism rather than by inducing cytotoxicity. In addition, NF-κB reporter assays demonstrated that RA attenuates TNF-α-induced transcriptional activation at higher concentrations, supporting its proposed anti-inflammatory properties and potential to modulate pro-tumorigenic signaling. Finally, our in silico studies predicted that RA may interact with proteins such as CAII, CES1, EGFR, and PLA2G2A, implicating it in the modulation of pathways related to proliferation and cell survival. Collectively, these findings suggest that RA may serve as a promising scaffold for the development of future anticancer agents, particularly in the context of colorectal cancer.

ZXGD Decoction, a traditional Chinese formulation historically used for cardiovascular ailments, was evaluated for its efficacy in coronary heart disease (CHD) through an integrated network pharmacology and randomized controlled trial (RCT) approach. Its selection was rooted in documented therapeutic benefits for blood stasis and endothelial dysfunction, with modern pharmacology identifying active compounds (e.g., luteolin, quercetin) targeting inflammation and oxidative stress pathways. Network analysis revealed ZXGD’s multi-target mechanism, prominently modulating the PI3K-AKT and NF-κB pathways, supported by robust molecular docking scores (binding affinity < -7.0 kcal/mol). These findings align with CHD pathophysiology, suggesting ZXGD disrupts critical inflammatory cascades. In a double-blind RCT (n = 180), ZXGD adjunct therapy significantly improved angina frequency (35% reduction vs. control, p < 0.01) and endothelial function (FMD increase: 2.8% ± 0.5 vs. 1.2% ± 0.4, p < 0.05) over 12 weeks, with no severe adverse events. This underscores ZXGD’s clinical potential as a safe complementary treatment. Notably, lipid profile enhancements (LDL-C reduction: 18.3% vs. 11.7%) correlated with predicted network targets, including LDLR and HMGCR. Our results bridge traditional use with mechanistic evidence, reinforcing ZXGD’s role in CHD management. While prior studies emphasize ZXGD’s anti-thrombotic effects, this work uniquely validates its anti-inflammatory and lipid-modulating properties, addressing gaps in understanding its systemic impact. Clinically, these findings advocate for ZXGD’s integration into CHD therapeutic protocols, particularly for patients with residual inflammatory risk.

The colonization and persistence of Pseudomonas aeruginosa in chronically diseased lungs are driven by various virulence factors. However, pulmonary infections in cystic fibrosis (CF) patients are predominantly polymicrobial. While Achromobacter xylosoxidans is an opportunistic pathogen in these patients, its impact on P. aeruginosa virulence during co-infection remains largely unknown. This study investigated P. aeruginosa interaction with two clonally related A. xylosoxidans strains, Ax 198 and Ax 200, co-isolated from CF sputum. We found that the interaction was strain-dependent, with Ax 200 significantly reducing P. aeruginosa virulence in a zebrafish model, providing the first in vivo evidence of this interaction. Proteomic analysis revealed that P. aeruginosa proteome was differently impacted by the two A. xylosoxidans strains, with Ax 200 altering proteins involved in biofilm formation, swimming motility, iron acquisition, and secretion systems. These findings were validated by phenotypic assays, confirming that A. xylosoxidans affected major P. aeruginosa virulence phenotypes, including biofilm formation, swimming motility, and siderophore production. Genetic analysis confirmed that distinct regulatory mechanisms, including iron cycle pathways, may account for the strain-dependent effects. These findings reveal a novel multi-target competitive mechanism through which A. xylosoxidans significantly disrupts P. aeruginosa virulence.

Traditional Chinese medicine (TCM), with its roots in centuries of clinical practice, has established itself as an effective therapeutic system that involves a diverse range of herbal plants. Despite its proven efficacy, the intricate relationships between herbal multi-component preparations and multi-target therapies present challenges for systematic study, thereby limiting its broader application in managing chronic diseases. In this work, we aim to identify novel therapeutic targets of natural compounds found in TCM herbs by leveraging advanced hypergraph representation learning techniques. Following the multi-component, multi-target pharmacological mechanisms, we first construct two hypergraphs to represent herb–compound and disease–target interactions, respectively. The connection between these hypergraphs is established through compound–target associations. A convolutional operator is then employed to capture the high-order correlations between compound (or target) nodes and herb (or disease) hyperedges within each hypergraph. Furthermore, we incorporate the PageRank algorithm and a multi-head attention mechanism to enhance the representation capabilities of node embeddings. By integrating these methods, our model is able to accurately identify novel therapeutic targets of natural compounds in TCM herbs in an end-to-end manner. Extensive experiments conducted on three benchmark datasets demonstrate the superior performance of our model when compared with several state-of-the-art approaches. Furthermore, case studies on two natural compounds, coumarin and progesterone, reveal that 7 and 8 out of the Top-10 identified targets, respectively, have been validated through literature review. These results highlight the effectiveness of our model in discovering new therapeutic targets for natural compounds in TCM.