This study aimed to combine network pharmacology with in vitro experiments to identify the key targets and potential mechanisms of salidroside (Sal) in the treatment of acute lung injury (ALI). Potential targets related to Sal and ALI were retrieved from the ChEMBL, SuperPRED, SwissTargetPrediction, GeneCards, OMIM, and CTD databases. Overlapping targets were imported into the STRING database and Cytoscape software to construct a protein-protein interaction (PPI) network and identify core targets. Functional enrichment analysis of these core genes, including GO and KEGG pathways, was performed using the DAVID database. Two genes, MAPK14 and GPX4, directly relevant to subsequent validation, were selected for molecular docking analysis. Furthermore, an in vitro model of ALI was established using LPS-induced alveolar type II epithelial cells to verify the protective mechanism of Sal. A total of 355 potential targets associated with Sal in ALI treatment were identified. In vitro experiments showed that, compared to the LPS group, the Sal group exhibited significantly reduced secretion of IL-6, ROS, p-MAPK, MDA, and Fe²⁺, along with increased GPX4 expression and attenuated lung injury. Integrated network pharmacology and experimental validation suggest that Sal pretreatment alleviates inflammatory response and oxidative stress, likely through regulation of the MAPK/GPX4 signaling pathway, thereby providing protection against lung tissue injury.

Inflammatory bowel disease (IBD), particularly ulcerative colitis, involves disruption of the intestinal mucosal barrier due to ecological and metabolic imbalances in the gut as its underlying pathology. Current therapies for Ulcerative colitis (UC) exhibit limited efficacy and adverse effects, necessitating the development of novel treatment strategies. Naringin and osthole are natural herbal compounds that show therapeutic potential in various inflammatory models due to their excellent anti-inflammatory activity. However, their combined therapeutic effects and precise mechanisms in UC remain unreported. This study aimed to explore the therapeutic effectiveness and mechanism of naringin combined with osthole in addressing dextran sodium sulfate (DSS)-induced colitis. The investigation centered on their impact on the disruption of the intestinal epithelial cell barrier, modulation of intestinal flora composition, alteration of metabolites, and inflammation model in vitro. Modal assessment encompassed body weight, disease activity index (DAI) score, colon length, and histopathological examination. Intestinal barrier integrity was evaluated through Quantitative Real-Time PCR, western blotting, and immunofluorescence staining. Microbiota abundance and metabolic levels wereassessed using 16S ribosomal RNA gene sequencing and metabolomics analysis. Protein expression levels ofpertinent pathways and associated receptors were testedthrough network pharmacology prediction and western blot analysis. Naringin and osthole synergistically relieved colitis symptoms in mice compared with either drug alone or 5-aminosalicylic acid, as evidenced by weight loss recovery, DAI scores, and colon length preservation. Mechanistically, naringin combined with osthole down-regulated the expression level of JNK/NF-κB signaling pathway related proteins and repaired intestinal barrier. Furthermore, the combination regulates the composition of the microflora and promotes the restoration of a steady state of the microflora.Metabolomicrevealed amino acid-tryptophan metabolism as a key metabolic pathway. It also reveals the microbiota-tryptophan pathway as a potential therapeutic strategy. Naringin combined with osthole can alleviate DSS-induced colitis more effectively by JNK/NF-κB signaling pathway, repairing barrier function and regulating intestinal microbiota and metabolites. These findings provide a theoretical basis for the combination therapy strategy to enhance the efficacy of potential functional food in treating ulcerative colitis.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by deficits in social interaction and comorbid symptoms including anxiety and cognitive problems. The main pathological mechanisms underlying ASD are synaptic abnormalities and neuroinflammation. Ginger, commonly used as a spice, has been reported to enhance neurogenesis and attenuate inflammation in neurological disease; however, its effects on ASD remain unknown. This study aimed to investigate the therapeutic effects and molecular mechanisms of ginger extract (GE) in ASD. Prenatally valproic acid (VPA)-exposed mice were orally administered GE for 4 weeks from 6 weeks of age. Behavioral tests were performed to assess social interaction, anxiety, and cognitive functions. Network pharmacology and molecular docking analyses were used to predict targets and mechanisms of GE in ASD, which were verified using western blotting. Histological changes, including neurogenesis, neuroinflammation, and synaptic formation, were analyzed using immunostaining, western blotting, and qRT-PCR. GE ameliorated VPA-induced social deficits, anxiety-like behavior, and memory impairments. Network pharmacology identified AKT as a core molecular target of GE, and its active compounds exhibited high binding affinity for AKT. Consistent with these predictions, GE increased AKT and GSK3β phosphorylation in the hippocampus of mice, thereby restoring neuronal development, as evidenced by the increased Ki67- and DCX-positive cells. GE also mitigated gliosis and reduced STAT3 phosphorylation and TNF-α upregulation, thereby suppressing neuroinflammation and synaptic loss. GE alleviates ASD-like behaviors by promoting neuronal and synaptic development while suppressing neuroinflammation through AKT/GSK3β signaling, highlighting its potential as a natural supplement for ASD prevention.

Epidemiological studies indicate that Metabolic Associated Fatty Liver Disease (MAFLD) has become the most prevalent chronic liver disease globally. Identifying bioactive molecules from natural plants that can prevent and ameliorate MAFLD is an effective approach to discovering new MAFLD therapeutics. Liupao tea possesses antioxidant and metabolic regulatory properties. This study aimed to identify the active components of Liupao tea polyphenol extract (PLE), and its potential targets and mechanisms of action in the prevention of MAFLD. We performed experiments on MAFLD mice to establish the prevention efficacy of PLE against MAFLD and determine its optimal dosage. Furthermore, network pharmacology, molecular docking, and SPR were performed to predict that PLE's targets for preventing MAFLD concentrate on the hepatic EGFR target. Knockout of EGFR in hepatocytes attenuated the effects of PLE. These findings indicate that PLE inhibits lipid accumulation by downregulating the EGFR/pEGFR-AKT/pAKT-SREBP-1-ACC1 pathway through binding to the EGFR receptor, thereby preventing MAFLD.

Thrombotic disorders remain a global health burden, necessitating novel antiplatelet agents with improved safety and efficacy. This study investigates the molecular mechanisms of two lignans, 6'-Hydroxyjusticidin B (6'-HJB) and Neojusticin A (Neo-A), isolated from Justicia procumbens L., through an innovative target-driven strategy integrating LC/MS, proteomics, network pharmacology, and biophysical validation. For the first time, integrin β3 (ITGB3) was identified as their direct molecular target, with microscale thermophoresis (MST) confirming high-affinity binding, the dissociation constant (Kd) = 0.0642 ± 0.005 μM for 6'-HJB; 0.0097 ± 0.001 μM for Neo-A. This study not only elucidates the structural basis of their activity-C-6 hydroxylation in 6'-HJB enhances ITGB3 specificity, whereas Neo-A’s fused furan ring optimizes COX-1 interaction, but also establishes a paradigm shift from phenotypic screening to target-validated natural product research. The findings position 6'-HJB and Neo-A as promising candidates for the development of safer, ITGB3-mediated antithrombotic therapies, with future efforts directed toward structural optimization and preclinical validation.Graphical

Exploring the Mechanism of Oral Cancer With Shikonin Based on the Network Pharmacology and Molecular Docking Technology.

As a traditional hair-growth-promoting herb, Platycladi Cacumen (PC) has a long history of folk application in the field of hair loss improvement. Preliminary modern pharmacological studies have suggested that its active components may exert potential effects by regulating hair follicle-related signaling pathways; however, for androgenetic alopecia (AGA), the exact targets and specific regulatory mechanisms of PC remain unelucidated, which provides a direction for research on natural drug-based intervention in AGA. In this study, network pharmacology was employed to predict the active components and core targets of PC. Targets associated with AGA were collected, and the intersection targets between PC and AGA were identified. Subsequently, protein-protein interaction (PPI) analysis, Gene Ontology (GO) enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed on the intersection targets to screen out the core targets. Thereafter, molecular docking and molecular dynamics(MD) simulation were conducted to validate the interactions between key active components and core targets. The component-target network diagram included 1044 interaction relationships between 32 components and 439 targets, among which quercetin(160), apigenin(150), myricetin(129), and hinokinin(105) were identified as key components. The disease-target network diagram summarized 410 targets associated with AGA. Through PPI network analysis, key targets such as ESR1(46), BCL2(44), INS(44), AR(42), and STAT3(40) were screened out. The results of GO enrichment analysis and KEGG pathway analysis revealed that PC may exert its effects by regulating the EGFR receptor molecule and pathways including the HIF-1 signaling pathway. Molecular docking results showed that the binding energies of all complexes were less than -5.0 kcal/mol, indicating favorable binding effects. MD simulation results showed that the root-mean-square deviation (RMSD), root-mean-square fluctuation (RMSF), radius of gyration (Rg), solvent-accessible surface area (SASA), two-dimensional free energy landscape (FEL-2D), and FEL-3D of the simulation system all remained in an equilibrium state with small fluctuation amplitudes. This result indicated that the molecular system had a stable overall conformation, restricted local residue movement, a compact spatial structure, and stable internal chemical bonds-collectively predicting that the quercetin-STAT3, apigenin-AR, myricetin-STAT3, and hinokinin-AR complexes may exhibit satisfactory binding stability. Collectively, Overall, this study systematically investigated the mechanism of action and potential value of PC leaves in intervening in AGA, providing a solid theoretical basis for the intervention of AGA with PC.

BackgroundDiabetic peripheral neuropathy (DPN) is a common complication of type 2 diabetes mellitus (T2DM) with limited treatment options. The traditional Chinese medicine Jinmaitong (JMT) has demonstrated efficacy in treating DPN in both clinical and animal studies. It is worth noting that macrophage polarization appears to play a significant role in the onset and progression of DPN. However, whether the specific mechanism by which JMT exerts its neuroprotective effects is related to macrophage polarization still requires further in-depth investigation.MethodsT2DM model was established using Sprague–Dawley (SD) rats induced by a high-fat diet for six weeks combined with streptozotocin (STZ) injection. After modeling and drug administration, the DPN status was assessed using the von Frey test to test mechanical threshold, the hot plate test and tail flick test to evaluate thermal response latency, and the bioelectric amplifier to measure motor nerve conduction velocity.In the first batch of in-vivo experiments (Batch 1), after establishing the type 2 diabetes model, we conducted herbal formula JMT administered daily via oral gavage for another four weeks, eight weeks or twelve weeks, with each study comprising four groups: control group (CON), DPN group (DPN), low-dose JMT (7.6 mg/kg) treated group (DPN + JMT), and high-dose JMT (15.2 mg/kg) treated group (DPN + JMTH). The pharmacological effects of JMT on neurological function, neuropathology, and the levels of M1 and M2 macrophage cytokine markers were evaluated in serum and sciatic nerve, respectively. After chemical profiling of JMT by liquid chromatography coupled with high-resolution mass spectrometry, network pharmacology analysis was subsequently employed to predict the potential signaling pathways that JMT targeted in treating DPN. We further explored JMT’s neuroprotective effect in a second batch of in-vivo experiments. To do this, we co-administered the JAK2/STAT3 inhibitor AG490 along with macrophage polarizing agents: LPS and interleukin-4 (IL-4). The changes of M1 and M2 macrophages in bone marrow was investigated by cytometry, while the macrophages in sciatic nerves were observed by immunofluorescence. Myelin morphology was observed with Luxel fast blue staining and transmission electron microscopy. Immunofluorescence was performed to evaluate nerve injury and regeneration, with S100 and neurofilament 160 (NF160) used to label Schwann cells and axons respectively in the sciatic nerve. The protein expressions of JAK2/STAT3 signaling in sciatic nerves were examined by Western blot.ResultsJMT significantly improved neurological function and pathological damage in type 2 DPN rats. Eight weeks after diabetes induction, DPN rats showed a significant increase in pro-inflammatory cytokines and a concurrent decrease in anti-inflammatory cytokines. JMT administration effectively restored the imbalance. Furthermore, JMT reduced the proportion of M1 macrophages while increasing that of M2 macrophages. JMT promoted the polarization of macrophages from the M1 to the M2 phenotype in both bone marrow-derived macrophages and those infiltrating the sciatic nerve, which was mediated through the suppression of abnormal activation of the JAK2/STAT3 signaling pathway.ConclusionsJMT promotes the polarization of macrophages from the M1 to M2 phenotype and alleviates neuroinflammation in T2DM rats with DPN, which is associated with inhibition of the JAK2/STAT3 signaling pathway. These findings highlight the neuroprotective potential of JMT through immunomodulatory mechanisms.Graphical Supplementary InformationThe online version contains supplementary material available at 10.1186/s13020-025-01300-0.

Background: Spinal cord injury (SCI) represents a form of traumatic damage to the central nervous system, and oligodendrocytes play a central role in SCI recovery. Ferroptosis is a major factor in the pathophysiological development of SCI symptoms. Pterostilbene (Pte) has antioxidant, anti-inflammatory, and neuroprotective effects. This study aims to investigate the potential role of Pte in SCI. Methods: A SCI model of rats was constructed. The BBB score assessment, the footprint test, EC staining, immunofluorescence (IF), and Western blot (WB) were conducted to observe the neuroprotective effects of Pte. The factors of ferroptosis, such as Glutathione (GSH), Malondialdehyde (MDA), Fe2+, solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4), were assessed. Then, transcriptomic data, network pharmacology, molecular docking analysis, and the erastin-induced ferroptosis model of OLN-93 cell lines were used to investigate the mechanism of inhibiting ferroptosis by Pte. Results: Pte treatment restored motor function and spinal cord tissue in SCI rats. Furthermore, Pte dramatically decreased oligodendrocyte ferroptosis. Finally, we discovered that Pte can repair SCI by blocking ferroptosis via the Keap1/Nrf2/SLC7A11/GPX4 axis. Conclusions: Pte reduces lipid peroxidation via the Keap1/Nrf2/SLC7A11/GPX4 axis, which reduces the development of ferroptosis in oligodendrocytes and improves locomotor function in rats with SCI.

Integrating UPLC-Q-TOF-MS/MS, feature-based molecular networking, network pharmacology, and molecular docking to investigate the mechanism of Persicae Ramulus against myocardial ischemia.

Network pharmacology and experimental validation identify the targets of Marsdenia tenacissima in neutrophilic asthma treatment.

Integrating transcriptomics and network pharmacology to investigate YangxinDingji Capsule alleviates myocardial injury in tachyarrhythmia rat.

Exploring the mechanism of BuNao AnShen Capsules in the treatment of insomnia based on network pharmacology and experimental validation.

The search for effective treatments for neurodegenerative diseases, particularly Alzheimer's disease, has been fraught with challenges. Alzheimer's disease accounts for 60-80% of dementia cases globally, affecting approximately about 50 million people. Currently, drug repurposing has emerged as a promising strategy in new drug development, attracting significant attention from regulatory agencies, such as the US FDA. This study aimed to investigate the potential therapeutic role of dolutegravir in Alzheimer's disease (AD) treatment using a novel network pharmacology approach. Specifically, it explored the interaction of dolutegravir with key molecular targets involved in AD pathology, predicted its effects on relevant biological pathways, and evaluated its viability as a new therapeutic candidate. This study employed a network pharmacology framework to evaluate dolutegravir, an antiretroviral drug, as a potential treatment for Alzheimer's disease, shedding light on its possible therapeutic mechanisms. A network pharmacology approach was used to predict the drug targets of dolutegravir. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to identify interacting pathways. Additionally, protein- protein interaction (PPI) network analysis was conducted to assess key interactions and molecular docking studies were performed to evaluate the binding affinity of dolutegravir to the predicted targets. PPI network analysis revealed that dolutegravir interacted with several key targets, including BRAF, mTOR, MAPK1, MAPK3, NOS1, BACE1, CAPN1, CASP3, CASP7, CASP8, CHUK, IKBKB, PIK3CA, and PIK3CD. KEGG pathway analysis suggested that dolutegravir could influence amyloid-beta formation, amyloid precursor protein metabolism, and the cellular response to amyloid-beta. Molecular docking results showed the highest binding affinity of dolutegravir for PI3KCD (-8.5 kcal/mol) and MTOR (-8.7 kcal/mol). The findings indicated that dolutegravir holds significant potential in modulating key pathways involved in Alzheimer's disease pathogenesis. These results provide a strong foundation for further investigations into the therapeutic efficacy and safety of dolutegravir in the treatment of Alzheimer's disease. The use of drug repurposing strategies, leveraging Dolutegravir's established pharmacological profile, offers a promising route for accelerated therapeutic development in AD.

Integrated strategy of serum metabolomics, network pharmacology and experimental validation on revealing the bioactive metabolites and mechanism of Lycium ruthenicum Murr. against colorectal carcinoma.

Tongmaijiangzhi formula and its hypolipidemic compounds attenuate hyperlipidemia and hepatic steatosis via AMPK/NF-κB pathway.

The total triterpenoid extract of Prunella vulgaris exerts anti-inflammatory effects by suppressing the NF-κB/TLR/TNF-α signaling axis in Lipopolysaccharide-stimulated RAW264.7cells.

Taikong Blue Lavender Essential Oil (TLEO) is derived from a proprietary space-bred cultivar of Lavandula angustifolia and cultivated under pristine conditions in Xinjiang, China. TLEO has long been used by regional people to treat various skin disorders such as hyperpigmentation, trans-epidermal water loss, collagen degradation, and poor wound healing. Despite the ethnopharmacological applications of TLEO, the molecular basis of its dermatological efficacy remains poorly defined. This study integrated network pharmacology, molecular docking, and invitro assays to systematically investigate how TLEO works against inflammatory skin conditions, focusing on its key compounds and biological targets. A total of 66 skin disorder-related genes were identified through network pharmacology, with gene enrichment analyses highlighting the TNF signaling pathway as a critical mediator. Protein-protein interaction analysis revealed MMP9, EGFR, and PTGS2 as core targets. Molecular docking confirmed that linalool and linalyl acetate, the primary constituents of TLEO, exhibited moderate binding affinities with these targets. Invitro experiments using TNF-α-stimulated HaCaT cells demonstrated that treatment with 0.01% TLEO significantly (p < 0.05) reduced oxidative stress markers (NO, ROS, MDA), restored antioxidant enzymes (SOD, CAT), and downregulated inflammatory cytokines (IL-6, IL-1β, IL-8). TLEO also inhibited the phosphorylation of p38 MAPK and NF-κB p65, suppressed PTGS2 and MMP9 expression, and restored EGFR levels, indicating anti-inflammatory and barrier-restorative functions. The study establishes a scientific foundation for the use of TLEO as a multifunctional ingredient in dermatological applications and highlights its value as a sustainable crop for regional economic development in Xinjiang.

Serum pharmacochemistry was applied to identify the blood-absorbed constituents of dry ginger (DG) and processed ginger (PG). Network pharmacology was employed to construct compound-target-pathway interaction networks, which were further validated through molecular docking to assess the binding affinities between ligands and key targets. Furthermore, a dextran sulfate sodium (DSS)-induced murine model of ulcerative colitis (UC) was established to systematically investigate the multi-target regulatory mechanisms of DG and PG. The processing of DG transforms gingerols into shogaols, thereby modifying its bioactive composition and physiological functions. DG, which is rich in 6-gingerol, mitigates intestinal inflammation through the inhibition of the PI3K-Akt signaling pathway, whereas PG increases the levels of shogaols and zingerone, promoting gut barrier restoration and reducing occult bleeding. Our findings illustrate how processing alters the functional properties of ginger, offering valuable insights for the development of ginger-derived products tailored to specific gastrointestinal health benefits.

Integrated multi-omics, network pharmacology, and experimental validation reveal that Yinmeikuijie decoction alleviates ulcerative colitis by inhibiting arachidonic acid metabolism.