Multi-target Mechanism Research Articles

Traditional Chinese medicine (TCM) has been used in the treatment of vascular cognitive impairment and dementia caused by chronic cerebral hypoperfusion (CCH) in patients for hundreds of years. Ethnopharmacological researches have been conducted in recent years to elucidate their therapeutic effects on cognitive deficits and potential mechanisms in animal models. This manuscript critically reviewed recent 5-year experimental researches from PubMed on the topic, including 11 TCM formulae, 8 herb extracts, and 21 pure compounds extracted from TCM, including polyphenols, flavonoids, alkaloids, terpenoids, saponins, iridoid glycosides, glucosides, and others in rodent CCH models, using bilateral common carotid artery occlusion (BCCAO, 2VO), bilateral common carotid artery stenosis (BCAS), and unilateral common carotid artery occlusion (UCCAO). The underlying mechanisms are multiple, including the maintenance of blood brain barrier and endothelium integrity, the increase in cerebral blood flow, the amelioration of white matter lesions, the modulation of microglia M1/M2 phenotype, the scavenge of reactive oxidative oxygen species and reduction of proinflammatory factors, the maintenance of mitochondrial function, the inhibition of apoptosis, ferroptosis and pyroptosis, and the promotion of neuronal regeneration and angiogenesis through the regulation of gene/protein expressions, including the Toll, NF-κB, MAPK, PPARγ, and/or Nrf2 pathways. These mechanisms are not mutually exclusive, rather they play an integrated role to fortify the multi-components, multi-targets feature of TCM in the treatment of CCH and human vascular cognitive impairments.

Matrine, a natural compound from traditional Chinese medicine, demonstrates anti-esophageal cancer (EC) efficacy via multi-target mechanisms. In this study, integrated network pharmacology and WGCNA identified MGLL, EPHX2, and CES2 as key targets for matrine. Clinical nomograms indicated their prognostic value, and bioinformatics analysis confirmed their downregulation in EC, correlating with tumor stage, survival, and immune infiltration. Molecular docking confirmed stable binding between matrine and these targets. In vitro experiments showed matrine upregulates their expressions, concurrently inhibiting EC cell proliferation and inducing apoptosis. In conclusion, matrine treats EC through dual actions-direct antitumor effects and immunomodulation-highlighting its therapeutic potential.

With the acceleration of globalization, traditional medicine, particularly Traditional Chinese Medicine (TCM), is progressively gaining increased global attention and recognition. TCM has not only achieved cross-border circulation in terms of product trade but has also become a significant vehicle for cultural exchange. Against the backdrop of growing global health demands, the unique efficacy and multi-target mechanisms of TCM provide broad potential for its application in areas such as chronic diseases and immune regulation. However, the internationalization of TCM still faces numerous challenges, including cultural differences, incomplete understanding of its pharmacological mechanisms, and the lack of global standardization. This paper examines the international context of TCM, its cross-cultural communication mechanisms, and the driving forces behind its application, analyzing the roles of academic exchanges, digital platforms, and multinational corporations in promoting the global dissemination of TCM. Furthermore, the article proposes key future directions for the international development of TCM, including the application of innovative technologies, cultural adaptation strategies, and interdisciplinary collaboration. Through these measures, TCM is poised to assume a more prominent position within the future global health system.

Protective effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside against cadmium toxicity involve BDNF/TrkB and PI3K/Akt signaling pathways.

Comorbidities, associated with higher rates of disability and mortality, have emerged as a global focus and a significant challenge. Psoriasis (PSO) and inflammatory bowel disease (IBD) are autoimmune disorders with frequent comorbidity and shared inflammatory pathways, yet targeted therapies remain limited. Utilizing an integrated approach combining network pharmacology, bioinformatics, machine learning, molecular docking, and in vitro experiments, we investigated Hansenia weberbaueriana (HW) for its potential to treat the comorbidity of PSO and IBD. Bioinformatics analysis identified 38 intersecting genes between PSO-IBD comorbidity and HW, predominantly enriched in the advanced glycation end-product (AGE)-receptor for AGE (RAGE) signaling pathway. Combining machine learning, weighted gene co-expression network analysis, and molecular docking prioritized PTGS2 (Cyclooxygenase 2 [COX2]) as the top target, with Cnidilin (HW8), a bioactive coumarin derivative from HW, exhibiting high-affinity binding (binding energy: -6.418kcal/mol and molecular mechanics-generalized born surface area: -35.43kcal/mol). Furthermore, 100-ns molecular dynamics (MD) simulations confirmed stable Cnidilin-PTGS2 interactions (root mean square deviation ∼2.3 Å). In vitro analysis demonstrated that Cnidilin (10 µM) significantly suppressed COX2, tumor necrosis factor-alpha, and nitric oxide in RAW 264.7 cells, supporting its multi-target mechanism. This study provided the first evidence supporting the therapeutic role of HW in PSO-IBD comorbidity via COX2 suppression and established a machine learning-driven framework for identifying natural compounds with dual therapeutic potential.

Formononetin (FMN), an isoflavone derived from Radix Astragali and red clover, has promising therapeutic potential for a wide spectrum of respiratory diseases, including acute lung injury (ALI), pulmonary arterial hypertension (PAH), chronic obstructive pulmonary disease (COPD), asthma, and pulmonary fibrosis (PF). Mechanistically, FMN alleviates oxidative stress, inflammation, and fibrotic remodelling by activating Nrf2/HO-1, inhibiting NF-κB, and modulating the activity of the TGF-β/Smad signalling pathway. Evidence from cellular and animal studies has shown that FMN attenuates lung injury, prevents vascular remodelling, and slows the progression of fibrosis. However, its clinical translation is hampered by poor solubility, rapid metabolism, and low oral bioavailability, which limit its therapeutic effectiveness. To overcome these challenges, novel delivery systems—such as albumin-based FMN nanoparticles (FMN@BSA nanoparticles)—have been developed to increase the stability, bioavailability, and pharmacological potency of FMN. Despite encouraging preclinical outcomes, further studies are needed to clarify upstream mechanisms and conduct rigorous clinical evaluations. This review highlights the potential of FMN as a novel therapeutic candidate for respiratory diseases by summarizing its mechanisms of action and underscoring the importance of advanced delivery strategies in facilitating its future clinical application.

Objective: This study aimed to systematically investigate the potential antibacterial mechanisms of ethacridine in the treatment of diabetic foot ulcers (DFUs) by integrating network pharmacology, molecular docking, and molecular dynamics simulation approaches. Methods: The potential targets of ethacridine were predicted using the SwissTargetPrediction and PharmMapper databases and subsequently converted to gene symbols via the UniProt database. DFU-related and antibacterial-related targets were retrieved from the GeneCards and OMIM databases. The overlapping targets among ethacridine, DFU, and antibacterial-related genes were identified as candidate therapeutic targets. A “drug–disease–target” network was constructed using Cytoscape, while protein–protein interaction (PPI) networks were built through the STRING database. GO and KEGG enrichment analyses were performed using R software. Molecular docking was conducted to evaluate the binding affinities between core compounds and hub targets. Furthermore, molecular dynamics (MD) simulation was applied to assess the binding stability of the top-ranked compound–target complex. Finally, RT-qPCR was conducted on wound edge tissue samples from DFU patients treated with ethacridine to experimentally validate the mRNA expression of predicted hub genes. Results: A total of 302 potential ethacridine-related targets, 4264 DFU-related targets, and 1942 antibacterial-related targets were identified. Intersection analysis revealed 105 common targets potentially involved in the antibacterial effects of ethacridine against DFU. PPI network analysis highlighted 10 hub targets, including AKT1, EGFR, SRC, HSP90AA1, and MMP9. GO enrichment indicated significant involvement in responses to reactive oxygen species, regulation of inflammatory responses, responses to lipopolysaccharide, and bacterial molecular patterns. KEGG pathway analysis identified 157 relevant pathways, including the lipid and atherosclerosis, TNF signaling, IL-17 signaling, and the AGE–RAGE signaling pathways in diabetic complications. Molecular docking demonstrated favorable binding affinities (all < −5.0 kcal/mol) between ethacridine and the hub targets, with the strongest binding observed between MMP9 and ethacridine (−9.8 kcal/mol). These docking results suggest possible interaction tendencies that may contribute indirectly to Ethacridine’s network-level regulatory effects, rather than direct binding to all targets in vivo. Molecular dynamics simulation further confirmed the stable interaction between MMP9 and ethacridine. RT-qPCR validation in clinical DFU tissue samples demonstrated expression trends of key genes consistent with in silico predictions. These results reflect transcriptional regulation consistent with pathway modulation predicted by the network analysis, rather than direct protein–ligand binding across all targets. Conclusion: Ethacridine may exert antibacterial effects against bacterial biofilms in DFU through multi-target and multi-pathway mechanisms. These findings highlight ethacridine’s translational potential as a safe, readily available, and mechanistically validated topical agent for the clinical management of biofilm-associated diabetic foot infections.

Parkinson's disease (PD) is a progressive neurodegenerative disorder marked by the loss of dopaminergic neurons, resulting in impaired motor control and cognitive decline. Despite current therapies, there remains a need for more effective and multifaceted treatment strategies. This study explores the neuroprotective potential of exogenous Creatine in a Haloperidol-induced PD rat model, with a focus on reducing elevated homocysteine levels and targeting key proteins involved in PD pathology. We proposed a multitarget therapeutic approach, aiming to inhibit ADORA2A, alpha-synuclein, and monoamine Oxidase B (MAO-B) using the same bioactive compound. RNA-seq data from publicly available datasets (BioProject: PRJNA608432; GEO: GSE145814) were analyzed to identify dysregulated proteins and pathways associated with neuroactive ligand-receptor interaction, olfactory transduction, and phototransduction. Using computational methods, including RNA Seq analysis, molecular docking, MMGBSA, and ADME/T analyses, we screened for potential common ligand. Creatine showed a higher binding affinity for the target proteins compared to Levodopa (primarily used medication against PD), suggesting its therapeutic potential. In vivo experiments assessed the efficacy of Creatine through behavioural testing, redox status evaluation, biochemical markers, and histological analysis. The results potentially suggest that Creatine ameliorates motor deficits, reduces oxidative stress, and protects neuronal integrity in PD rats. Overall, our findings indicate that Creatine acts as a promising multitargeted approach, capable of modulating several PD-associated molecular mechanisms. This approach may further be validated and pave the way for more effective, broad-spectrum treatments for Parkinson's disease, addressing both its motor and non-motor symptoms.

We aimed to integrate network pharmacology, metabonomics, and fecal transcriptomics to elucidate the mechanism of action of Diyu Shengbai Tablets (DST) in leukopenia treatment. Key bioactive components and core targets of DST in leukopenia were identified using network pharmacology. Spleen metabonomics, fecal transcriptomics, and MetOrigin analysis were employed to investigate potential mechanisms. Molecular docking, molecular dynamics simulations, and enzyme-linked immunosorbent assay were performed to validate findings. Network pharmacology predicted Hypericin and Quercetin-3-galactoside-7-glucoside as major active constituents of DST. metabonomic analysis identified alterations in 133 differential metabolites in the spleen, with linoleic acid metabolism as the primary affected pathway. 16S rRNA sequencing indicated normalization of Lactobacillus abundance. Molecular docking and dynamics simulations showed low binding free energies between active compounds (e.g., Hypericin) and targets such as PIK3CA and SRC. Serum levels of Mouse interleukin-6 and Granulocyte colony-stimulating factor were elevated following DST intervention. DST modulates spleen metabolic profiles in leukopenic mice, restores gut microbial composition, and exerts therapeutic effects by regulating linoleic acid metabolism and influencing the PI3K-Akt signaling pathway. These findings provide a foundation for further exploration of multi-target mechanisms in traditional Chinese medicine for leukopenia management.

Berberine (BBR), a benzylisoquinoline alkaloid isolated from Chinese herb Coptis chinensis, has been widely used clinically to treat intestinal infectious diseases. Recently, it has been found to have multiple pharmacological effects, including anti-inflammatory activity and immune effects in inflammatory bowel disease (IBD). However, its exact targets remain to be elucidated. In this study, we used a mouse intestinal organoid–macrophage co-culture model to investigate the anti-inflammatory effects and immune effects of BBR. Our findings demonstrated that lipopolysaccharide (LPS) induced more robust inflammatory responses and epithelium damage in the co-culture system compared to the organoid alone. BBR effectively attenuated inflammation and restored epithelial barrier integrity by suppressing M1 macrophage polarisation and infiltration, alongside upregulating the expression and organisation of tight junction protein zonula occludens-1 (ZO-1). RNA sequencing and proteomic analysis revealed that BBR disrupted organoid–macrophage interaction by inhibiting chemokine (e.g., C-X-C motif chemokine ligand 1 (CXCL1) and macrophage migration inhibitory factor (MIF)) release from epithelial cells, thereby reducing macrophage recruitment. Collectively, our study establishes the organoid–macrophage co-culture system as a more physiologically relevant model for studying epithelial–immune interactions and elucidates the multi-target mechanism of BBR, which concurrently modulates epithelial cells, macrophages, and their crosstalk. These findings lay the foundation for further exploration of the therapeutic potential of BBR in inflammatory bowel disease and the development of targeted therapies that regulate cell interactions.

Alzheimer's disease (AD) is a progressive neurodegenerative disease and a leading cause of dementia, profoundly affecting individuals worldwide. However, there are limited therapeutic approaches to treat AD, with current approaches primarily focused on providing symptomatic relief rather than halting the disease progression, highlighting the need for alternative strategies. Nutraceuticals, which are derived from natural sources, offer a promising alternative because of their multi-targeted mechanisms that modulate inflammation, oxidative stress and neurodegeneration, addressing key symptoms of AD pathology. Studies from clinical trials state the challenges of bioavailability and efficacy of nutraceuticals. Advancements and refinements in food processing and delivery methods, along with personalized nutrition, offer a possible approach to overcome these drawbacks. Future directions rely on combination therapies incorporating nutraceuticals to enhance the bioavailability and efficacy of these compounds. This approach could pave way for innovative solutions of elevating nutraceuticals from farm to pharma that offer sustainable solutions to mitigate AD globally.

Research progress on the role of oxidative stress in the pathogenesis of vascular dementia and its treatment.

BackgroundAlzheimer's disease is a progressive neurodegenerative disorder marked by amyloid-β (Aβ) plaque deposition and neurofibrillary tangles composed of hyperphosphorylated tau. Dysregulation of glycogen synthase kinase-3β (GSK3β) promotes tau hyperphosphorylation and amplifies Aβ-induced neurotoxicity, driving pathogenesis. Despite extensive research, current therapies targeting these core mechanisms remain largely ineffective at halting disease progression.ObjectiveBased on prior clinical and preclinical evidence, we hypothesize that cannabidiol (CBD), a non-psychoactive phytocannabinoid, may exert multitarget therapeutic effects in AD by modulating Aβ aggregation, tau hyperphosphorylation, and GSK3β activity.MethodsWe investigated CBD's interactions with Aβ-42/40, tau, and GSK3β using molecular docking, molecular dynamics simulations and ADMET predictions.ResultsOur results show that CBD binds to Aβ with binding free energies of -7.81 kcal/mol, -7.46 kcal/mol, and -7.25 kcal/mol, disrupting aggregation by interacting with key residues (HIS6, HIS13, HIS14, GLU14, GLU22, ASP15, and ASP23). MD simulations confirm that CBD destabilizes Aβ's β-sheet structure, preventing fibril formation. CBD binds tau with binding free energies of -9.91 kcal/mol, -9.70 kcal/mol, and -9.66 kcal/mol, disrupting tau aggregation and preventing neurofibrillary tangle formation. MD simulations show that CBD induces structural changes in tau, reducing β-sheet packing and inhibiting tau-tau interactions. CBD also binds to GSK3β with binding energies of -8.94 kcal/mol, -8.51 kcal/mol, and -8.41 kcal/mol, competing with ATP to inhibit its kinase activity and reduce tau phosphorylation. ADMET analysis indicates CBD's favorable oral bioavailability and low toxicity.ConclusionsThese findings support CBD as a promising multitarget therapeutic for AD, warranting further preclinical and clinical investigations.

Plant-derived alkaloids exhibit significant anticancer potential, yet their multi-target mechanisms, spanning signaling pathways, programmed cell death, immunity, and metabolism, remain fragmented. This narrative review synthesizes recent preclinical evidence on five representative alkaloids: dendrobine (DDB), aloperine (ALO), levo-tetrahydropalmatine (L-THP), solamargine (SM), and cyclovirobuxine D (CVB-D). Using a dual-framework of compound-specific analysis and key regulatory modules (NF-κB, MAPK, PI3K/AKT/mTOR, JAK/STAT; apoptosis, autophagy, ferroptosis; immune checkpoints; metabolism/microbiota), the study identified convergent anticancer mechanisms with translational relevance. These alkaloids consistently suppress NF-κB, PI3K/AKT/mTOR, and MAPK pathways, and modulate JAK/STAT signaling. They induce apoptosis and ferroptosis, and block autophagic flux. Notably, EVO and SM downregulate PD-L1 via the MUC1-C/NF-κB/c-MYC axis, enhancing CD8⁺ T cell function. L-THP activates AMPK and remodels tumor metabolism. These mechanistic insights support rational co-therapies such as L-THP plus metabolic inhibitors, or ALO combined with bispecific immune checkpoint inhibitors. Overall, these alkaloids demonstrate systemic, multi-pathway anticancer efficacy, and represent promising partners in precision combination therapy. Clinical translation should prioritize formulation and pharmacokinetic optimization, biomarker-guided stratification, and preclinical validation of synergistic regimens.

In the context of clinical treatment for mixed vaginal infections, which pose dual challenges of biofilm resistance and immune microenvironment imbalance, this study synthesized 12 “triazole–Schiff base” hybrid compounds using previously reported synthetic routes. This design was guided by the core mechanism in which CYP24A1 enzyme overexpression in the vitamin D–vitamin D receptor immunoregulatory pathway leads to degradation of active vitamin D. Among them, compound 3c exhibited excellent broad-spectrum antibacterial and antifungal activity (MIC = 16 μg mL−1), low cytotoxicity, and significant inhibition and eradication of Candida albicans biofilms. Mechanistic studies revealed that the compound possesses both membrane-targeting disruptive effects and CYP24A1 inhibitory activity. At the enzymatic level, compound 3c achieved 32% inhibition of CYP24A1 at 100 μM. In cellular models, it did not affect basal CYP24A1 mRNA expression at 10−7 M, but when combined with 1,25(OH)2D3, it up-regulated CYP24A1 mRNA levels approximately twofold. Further LC-MS/MS analysis confirmed that the addition of 10−7 M of compound 3c significantly slowed the metabolic clearance of 1,25(OH)2D3 in HEK293T cells, maintaining an average concentration of 495 pM after 24 h. In summary, compound 3c employs a multi-target synergistic mechanism of “direct antimicrobial activity–immunomodulation–biofilm penetration” to effectively overcome biofilm-mediated resistance and improve the local immune microenvironment, thus offering a promising lead compound with clinical translational potential for addressing the therapeutic challenges in mixed infections.

We aimed to conduct a systematic analysis of the multi-target regulatory mechanisms of Ziyu glycoside I (Ziyu I) in cervical cancer progression using network pharmacology and molecular docking techniques. Additionally, we validated our findings using cellular experiments and animal models. The PhamMapper database was used to identify drug targets for Ziyu I, while cervical cancer-related disease targets were obtained from the DisGeNET, DrugBank, GeneCards, and OMIM databases. In vitro, the effects of Ziyu I on the proliferation, migration, apoptosis, and cell cycle were assessed using cultured HeLa and SiHa cervical cancer cells. Network pharmacology analysis identified 68 potential targets of Ziyu I in cervical cancer. Molecular docking findings revealed that Ziyu I had a stable binding ability to MAPK1, MAPK8, and MAPK14, which are key targets in the MAPK signaling pathway. Cellular experiments revealed that Ziyu I significantly inhibited the proliferation and migration of cervical cancer cells and promoted apoptosis and cell cycle arrest. Finally, a nude mice-loaded tumor assay revealed that Ziyu I had significant anti-cervical cancer activity in vivo. Ziyu I inhibits cervical cancer progression through a multitarget regulatory network involving MAPK signaling pathway-mediated inhibition of cell proliferation and migration, apoptosis activation, and cell cycle arrest.

Psoralen alleviates ulcerative colitis by suppressing inflammation, modulating oxidative stress, and regulating ferroptosis.

Background/Objectives: Glioblastoma (GBM), the most aggressive primary malignant brain tumor, has a dismal prognosis and limited treatment options. The dried rhizome of Ligusticum chuanxiong Hort. (Chuanxiong, CX) is a traditional Chinese medicinal herb frequently prescribed in formulas intended to invigorate blood circulation. CX also exhibits anti-glioma activity, but its molecular mechanisms remain incompletely understood. Methods: In this study, we combined transcriptomics and Raman spectroscopy to investigate the effects of reconstituted CX-dispensing granules (hereafter referred to as CXG solution) on U87MG cells, suggesting their dual role in promoting cell death and modulating collagen deposition and lipid metabolism. Results: Mechanistically, we demonstrated that the CXG solution downregulates hsa-miR-10a-5p, which directly targets BCL2L11, known to induce pro-apoptotic effects, as validated by qPCR and dual-luciferase reporter assays. Furthermore, the CXG solution and hsa-miR-10a-5p suppress lipid metabolism through a coherent feed-forward loop via targeting transcription factors SREBF1 and E2F1. An electrophoretic mobility shift assay (EMSA) confirmed E2F1 binds to the hsa-miR-29a promoter, leading to the synergistic repression of hsa-miR-29a-3p by SREBF1 and E2F1. Network pharmacology analysis combined with molecular docking suggested that the ferulic acid and adenosine in CX potentially modulate EGFR-the E2F1-hsa-miR-10a-5p axis. Conclusions: These findings elucidate CX’s multi-target anti-GBM mechanisms and propose a novel therapeutic strategy combining metabolic intervention with miRNA-targeted therapy, providing novel insights into feed-forward loop regulation in miRNA networks.

Shenmai injection attenuates sepsis-associated acute lung injury by remodeling gut microbiota and restoring steroid hormone biosynthesis.

Bioactive compounds and multitarget action mechanism of Erzhi pills in alleviating metabolic dysfunction-associated steatohepatitis.