Elucidating the Mechanism of Xiaoqinglong Decoction in Chronic Urticaria Treatment: An Integrated Approach of Network Pharmacology, Bioinformatics Analysis, Molecular Docking, and Molecular Dynamics Simulations.

Alzheimer's disease (AD) is a degenerative brain disease without a cure. Lonicerae Japonicae Flos (LJF), a traditional Chinese herbal medicine, possesses a neuroprotective effect, but its mechanisms for AD are not well understood. This study aimed to investigate potential targets and constituents of LJF against AD. Network pharmacology and bioinformatics analyses were performed to screen potential compounds and targets. Gene Expression Omnibus (GEO) datasets related to AD patients were used to screen core targets of differential expression. Gene expression profiling interactive analysis (GEPIA) was used to validate the correlation between core target genes and major causative genes of AD. The receiver operating characteristic (ROC) analysis was used to evaluate the predictive efficacy of core targets based on GEO datasets. Molecular docking and dynamics simulation were conducted to analyze the binding affinities of effective compounds with core targets. Network pharmacology analysis showed that 112 intersection targets were identified. Bioinformatics analysis displayed that 32 putative core targets were identified from 112 intersection targets. Only eight core targets were differentially expressed based on GEO datasets. Finally, six core targets of MAPK8, CTNNB1, NFKB1, EGFR, BCL2, and NFE2L2 were related to AD progression and had good predictive ability based on correlation and ROC analyses. Molecular docking and dynamics simulation analyses elucidated that the component of lignan interacted with EGFR, the component of β-carotene interacted with CTNNB1 and BCL2, the component of β-sitosterol interacted with BCL2, the component of hederagenin interacted with NFKB1, the component of berberine interacted with EGFR and BCL2, and the component of baicalein interacted with NFKB1, EGFR and BCL2. Through a comprehensive analysis, this study revealed that six core targets (MAPK8, CTNNB1, NFKB1, EGFR, BCL2, and NFE2L2) and six practical components (lignan, β-carotene, β-sitosterol, hederagenin, berberine, and baicalein) were involved in the mechanism of action of LJF against AD. Our work demonstrated that LJF effectively treats AD through its multi-component and multi-target properties. The findings of this study will establish a theoretical basis for the expanded application of LJF in AD treatment and, hopefully, can guide more advanced experimental research in the future.

Fufang Honghua Buji (FHB) granules, have proven efficacy against vitiligo in long-term clinical practice. However, its major active chemical components and molecular mechanisms of action remain unknown. The purpose of this study was to confirm the molecular mechanism of FHB's therapeutic effect on vitiligo utilizing network pharmacology, molecular docking, and molecular dynamics simulation prediction, as well as experimental verification. Traditional Chinese Medicine Systems Pharmacology (TCMSP) and HERB databases were used to obtain the chemical composition and action targets of FHB. Online Mendelian Inheritance in Man (OMIM), DrugBank, DisGeNET, GeneCards, and Therapeutic Target Database (TTD) databases were applied to screen for vitiligo-related targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed through the Matascape database. Molecular docking and dynamics simulation methods were for the analysis of the binding sites and binding energies between the FHB's active components and the targets. Finally, a vitiligo mouse model was created, and the therapeutic effect and molecular mechanism of action of FHB were validated using enzyme linked immunosorbent assay (ELISA), western blot (WB), and quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Additionally, hematoxylin-eosin staining (HE) and blood biochemical assays were conducted to assess the biosafety of FHB. The screening of chemical composition and targets suggested that 94 genetic targets of FHB were associated with vitiligo. The bioinformatics analysis suggested that luteolin, quercetin, and wogonin may be major active components, and nuclear factor-kappa B p65 subunit (RELA), signal transducer, and activator of transcription (STAT) 3 and RAC-alpha serine/threonine-protein kinase (AKT) 1 may be potential targets of FHB-vitiligo therapy. Molecular docking and dynamics simulation further demonstrated that luteolin, quercetin, and wogonin all bound best to STAT3. Through experimental verification, FHB has been demonstrated to alleviate the pathogenic characteristics of vitiligo mice, suppress the JAK-STAT signaling pathway, reduce inflammation, and increase melanogenesis. The in vivo safety evaluation experiments also demonstrated the non-toxicity of FHB. FHB exerts anti-inflammatory and melanogenesis-promoting effects via the effect of multi-component on multi-target, among which the JAK-STAT pathway is a validated FHB-vitiligo target, providing new ideas and clues for the development of vitiligo therapy.

Multi-omics and experimental validation reveal the mechanism of DanxiaTiaoban decoction in treating atherosclerosis.

Background: Osteoporosis is a prevalent bone metabolism disease characterized by a reduction in bone density, leading to several complications that significantly affect patients’ quality of life. The Achyranthes bidentata–Dipsacus asper (AB–DA) herb pair is commonly used in Traditional Chinese Medicine (TCM) to treat osteoporosis. This study aimed to investigate the therapeutic compounds and potential mechanisms of AB–DA using network pharmacology, molecular docking, molecular dynamics simulation, and experimental verification.Methods: Identified compounds of AB–DA were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Traditional Chinese Medicine Information Database (TCM-ID), TCM@Taiwan Database, BATMAN-TCM, and relevant literature. The main bioactive ingredients were screened based on the criteria of “OB (oral bioavailability) ≥ 30, DL (drug-likeness) ≥ 0.18.” Potential targets were predicted using the PharmMapper and SwissTargetPrediction websites, while disease (osteoporosis)-related targets were obtained from the GeneCards, DisGeNET, and OMIM databases. The PPI network and KEGG/GO enrichment analysis were utilized for core targets and pathway screening in the STRING and Metascape databases, respectively. A drug–compound–target–pathway–disease network was constructed using Cytoscape software to display core regulatory mechanisms. Molecular docking and dynamics simulation techniques explored the binding reliability and stability between core compounds and targets. In vitro and in vivo validation experiments were utilized to explore the anti-osteoporosis efficiency and mechanism of sitogluside.Results: A total of 31 compounds with 83 potential targets for AB–DA against osteoporosis were obtained. The PPI analysis revealed several hub targets, including AKT1, CASP3, EGFR, IGF1, MAPK1, MAPK8, and MAPK14. GO/KEGG analysis indicated that the MAPK cascade (ERK/JNK/p38) is the main pathway involved in treating osteoporosis. The D–C–T–P–T network demonstrated therapeutic compounds that mainly consisted of iridoids, steroids, and flavonoids, such as sitogluside, loganic acid, and β-ecdysterone. Molecular docking and dynamics simulation analyses confirmed strong binding affinity and stability between core compounds and targets. Additionally, the validation experiments showed preliminary evidence of antiosteoporosis effects.Conclusion: This study identified iridoids, steroids, and flavonoids as the main therapeutic compounds of AB–DA in treating osteoporosis. The underlying mechanisms may involve targeting core MAPK cascade (ERK/JNK/p38) targets, such as MAPK1, MAPK8, and MAPK14. In vivo experiments preliminarily validated the anti-osteoporosis effect of sitogluside. Further in-depth experimental studies are required to validate the therapeutic value of AB–DA for treating osteoporosis in clinical practice.

The bioactive components of Astragalus membranaceus and Salvia miltiorrhiza improved cardiac and renal function in chronic heart failure (CHF) and chronic kidney disease (CKD), respectively. However, the common regulating molecular mechanisms remain unclear. The aim of this study was to investigate these mechanisms using bioinformatics, network topology, and molecular dynamics simulation techniques. The active components and target sites of A. membranaceus and S. miltiorrhiza were obtained from the Traditional Chinese Medicine Systems Pharmacology database. The targets of CKD and CHF were obtained from various databases for a protein-protein interaction analysis. The Gene Ontology (GO) function and Kyotoencyclopedia of genes and genomes (KEGG) pathway enrichment of intersection targets were analyzed by using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) database. Molecular docking and dynamic simulations were conducted on the core ingredients and targets. The diagnostic efficiency of the key targets was evaluated by using receiver-operating characteristic (ROC) curves. A total of 70 active ingredients and 158 common targets were found. The top five core targets were AKT1, STAT3, TP53, MAPK1, and RELA. The GO enrichment analysis included apoptosis and oxidative stress. The KEGG pathway enrichment results indicated that the drug pair regulated the AGE-receptor for AGE signaling pathway, fluid shear stress and atherosclerosis, and the IL-17 signaling pathway. Molecular docking and dynamic simulations confirmed that the core ingredients had good affinity and stability with the key targets. The ROC curves confirmed the accuracy of every key target for identifying CKD and CHF and demonstrated that combining them improves diagnosis. The combination of A. membranaceus and S. miltiorrhiza proved effective for the treatment of CKD and CHF through various components, targets, and mechanisms. Moreover, it may predict the diagnostic value of key targets, providing a reference for clinical diagnostic applications.

Epimedii herba (EH) showed numerous activities and has the potential to treat periodontitis. However, the pharmacological mechanism has not been exhaustively elucidated. This study predicted the specific targets and mechanisms of EH to prevent and treat periodontitis. A traditional Chinese medicine system pharmacology database and analysis platform was used to screen key compounds of EH and their corresponding targets. Therapeutic Target Database and Comparative Toxicogenomics Database were used to identify targets related to periodontitis. Intersection targets were observed using a Venn diagram. The key components and corresponding protein targets of EH were searched. The intersection targets were obtained and then they were imported into the STRING database to construct a PPI network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. Molecular docking between the screened chemical components of EH and key targets was performed using Discovery Studio 2019. The binding stability between components and target proteins was confirmed using molecular dynamics simulations. The binding stability between components and target proteins was confirmed using molecular dynamics simulations. Through network pharmacological analysis, 23 active compounds of EH were identified, including kaempferol and icariin. Based on GeneCards, GEO, and other databases, 3291 periodontitis-related genes were obtained. Venn diagram analysis revealed 137 intersection targets of EH and periodontitis, and Protein kinase B (AKT1) and Tumor necrosis factor (TNF) were identified as the key targets of EH for periodontitis treatment. GO and KEGG analyses revealed that the primary pathways mediating the therapeutic effects of EH were related to cancer, lipid, and atherosclerosis. Molecular docking showed that 8-isopentenyl-kaempferol had the best binding ability to ESR1, which was confirmed by dynamics simulations. This study demonstrated that EH can be used for periodontitis treatment, and the corresponding targets and potential mechanisms were investigated based on network pharmacology, molecular docking, and dynamics simulation analysis. Notably, 8-isopentenyl-kaempferol exhibited good binding affinity and stability to ESR1, which may partially explain the molecular mechanisms of EH for treating periodontitis. Hence, EH can be a novel choice for the clinical treatment of periodontitis in the future.

Given the increasing incidence and disability rate of rheumatoid arthritis (RA) year by year, RA has become a common cause of disability. Danggui Niantong Decoction (DGNTD) has been shown to have therapeutic effects on RA. However, to date, its bioactive components and potential targets remain unclear. To systematically explore the potential mechanisms of DGNTD in the treatment of RA, we utilized a combination of network pharmacology, Mendelian randomization, molecular docking, and molecular dynamics simulation. We used the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database to identify DGNTD’s active ingredients and potential targets, and GeneCards to screen RA-related targets. Common targets were identified via Venn analysis. We built a protein–protein interaction network and a drug-ingredient-target map with Cytoscape. Based on shared targets, we conducted gene ontology and Kyoto encyclopedia of genes and genomes enrichment analyses to reveal key pathways. To validate results, we used R for Mendelian randomization to assess core targets’ correlation with RA. We used Autodock for molecular docking and GROMACS for dynamics simulations to verify complex stability. DGNTD contains 316 active ingredients and 276 potential targets. Through protein–protein interaction network analysis and drug-ingredient-target network screening, we identified 215 active ingredients and 200 common targets, with quercetin and naringenin as core active ingredients. The core targets included RAC-alpha serine/threonine-protein kinase, cellular tumor antigen p53, signal transducer and activator of transcription 3, mitogen-activated protein kinase 1, mitogen-activated protein kinase 3, and Myc proto-oncogene protein. The Kyoto encyclopedia of genes and genomes and gene ontology enrichment analyses revealed 186 signaling pathways, with core targets mainly involving the PI3K/AKT pathway, lipid metabolism, and atherosclerosis-related biological processes. After validation through Mendelian randomization, RAC-alpha serine/threonine-protein kinase and mitogen-activated protein kinase 3 may be key targets for the treatment of RA by DGNTD. The molecular docking and dynamics simulations showed that the complexes formed by the core active ingredients and key targets exhibited strong stability. DGNTD exerts its therapeutic effects on RA through the synergistic action of multiple components, targets, and pathways, with its anti-inflammatory effects and potential molecular mechanisms being particularly noteworthy and warranting further investigation.

Deciphering the pharmacological mechanisms of Shenlingbaizhu formula in antibiotic-associated diarrhea treatment: Network pharmacological analysis and experimental validation

This study investigated the mechanism of action of Fibraurea recisa Pierre (FRP) in the treatment of chronic urticaria (CU) using a rat model and combinatorial analysis of network pharmacology, metabolomics, and molecular dynamics and dynamics simulation data, providing a rationale for its clinical use. Twenty-four Sprague-Dawley rats were categorized into control, model, high-dose FRP (40mg/kg body weight), and low-dose FRP (20mg/kg body weight) groups. The CU model was induced by ovalbumin. Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC/MS) was used to estimate the levels of various components in FRP. The rats in different groups were evaluated for scratching behavior, histopathological changes in the skin tissues based on hematoxylin/eosin staining, and the levels of inflammatory factors and indicators of mast cell degranulation. Metabolomics, network pharmacology, molecular docking and dynamics simulation, and Western blotting were used to analyze the mechanism of action of FRP. We identified 2,206 compounds in FRP based on UPLC/MS data analysis. Our data showed that the main active components in FRP were palmatine, jatrorrhizine, and coclaurine. FRP administration significantly reduced the scratching frequency, pathological characteristics of skin tissues, levels of inflammatory factors, and the degree of mast cell degranulation. Based on the combined analysis of metabolomics and network pharmacology data, phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) signaling pathway was identified as the key target of FRP. Molecular docking and molecular dynamics simulation demonstrated strong and stable binding of Akt with palmatine, jatrorrhizine, and coclaurine. Western blotting confirmed that FRP increased the levels of p-Akt and p-PI3K in skin tissue within the CU model. FRP significantly alleviated the symptoms and pathological changes of CU by modulating inflammation through upregulation of the PI3K-Akt signaling pathway.

The incidence of Polycystic Ovary Syndrome (PCOS) is increasing annually. This study aims to investigate the therapeutic mechanisms of Yikang Decoction (YKD) in the treatment of PCOS through the integration of GEO datasets, network pharmacology, and dynamic simulation. Active ingredients of YKD and their targets were collected from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform. Disease-relevant targets for PCOS were retrieved from several databases, including GeneCards, OMIM, PharmGKB, DrugBank, and GEO. The underlying pathways associated with the overlapping targets between YKD and PCOS were identified using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The mechanisms of interaction between the core targets and components were further explored through molecular docking and molecular dynamics simulations (MD). 139 potential active components and 315 targets of YKD were identified. A topological analysis of the PPI network revealed 10 core targets. These targets primarily participated in the regulation of biological processes, including cell metabolism, apoptosis, and cell proliferation. The pathways associated with treating PCOS encompassed PI3K-Akt signaling pathway, Lipid and atherosclerosis, MAPK signaling pathways, and Endocrine resistance signaling pathways. Moreover, molecular docking and MD have been shown to reveal a good binding capacity between active compounds and screening targets. This study systematically investigates the multi-target mechanisms of YKD in the treatment of PCOS, with preliminary verification provided through molecular docking and MD. The findings offer compelling evidence supporting the efficacy of YKD in treating PCOS.

Objective: Primary dysmenorrhea (PD) is a prevalent gynecological disorder. Yuanhu Zhitong oral liquid (YHZT) presents a promising alternative treatment for PD. However, the mechanisms underlying its efficacy remain unclear. This study aims to investigate the potential targets and mechanisms of action of YHZT in treating PD using network pharmacology, molecular docking, and molecular dynamics simulations. Methods: Potential compounds from YHZT were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the Traditional Chinese Medicine Information Database (TCMID). The relevant targets of these compounds were identified using the similarity ensemble approach (SEA) and the Swiss Target Prediction database. PD-related targets were retrieved from the Genecards, DrugBank, and Disgenet databases. ClusterProfiler was utilized for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. The compound-target-pathway (CTP) network was constructed to facilitate the identification of key compounds, core targets, and signaling pathways. Finally, molecular docking and molecular dynamics simulations were performed to evaluate the interactions between the targets and compounds. Results: A total of 153 putative compounds and 129 targets of YHZT were identified. Network topology analysis revealed eight core targets and six key compounds. The effects of YHZT were mediated by genes associated with hormone and steroid metabolism, as well as pathways involved in steroid hormone biosynthesis and cytochrome P450 enzymes. Docking results showed free binding energies ranging from −6.06 to −10.85 kcal/mol, indicating strong binding affinity between the compounds and targets. Molecular dynamics simulation results further confirmed the stability of these interactions. Conclusion: This study demonstrates that YHZT treats PD by suppressing inflammatory reactions and modulating hormone and cytochrome P450 concentrations. Key compounds such as ferulic acid, (R)-canadine, (S)-canadine, canadine, and asristolone are implicated in this process. These findings offer insights into the mechanisms underlying the action of YHZT and provide a foundation for further research in this area.

Modern medical practice has confirmed the efficacy of Mahuang Fuzi Xixin Decoction (MHFZXXD) in treating elderly bronchial asthma, but its specific mechanisms of action remain to be clarified. Therefore, this study utilizes network pharmacology, molecular docking techniques, and molecular dynamics simulations to explore the key active components, core target genes, and potential mechanisms of MHFZXXD in the treatment of elderly bronchial asthma. Active components and related targets of MHFZXXD were identified through the retrieval and screening of the TCMSP, Swiss Targets Prediction, and Uniprot databases. Relevant targets for elderly bronchial asthma were searched using the GeneCards, OMIM, and Pharm GKB databases, followed by the selection of intersecting targets between the drug’s active components and the disease. A PPI network diagram was created using String and Cytoscape software, and the intersecting targets of the disease and the active components of traditional Chinese medicine were imported into the DAVID database for GO and KEGG enrichment analysis to further explore their potential mechanisms of action. Subsequently, molecular docking and molecular dynamics simulations were performed using AutoDock Vina and Gromacs to verify the binding capacity and stability of the core genes with the key active components. The study results indicate that the active components of MHFZXXD, such as quercetin, luteolin, and kaempferol, target multiple genes including AKT1, EGF, MYC, TGFB1, PTEN, and CCND1. They exert effects through signaling pathways such as TNF, PI3K-Akt, and HIF-1. Molecular docking and dynamics simulations show that the core targets bind stably with the key active components. Overall, MHFZXXD may reduce inflammatory responses and improve hypoxic conditions and apoptosis during the progression of elderly bronchial asthma through multiple active components, targets, and signaling pathways, thereby delaying the malignant progression of the disease. This provides relevant evidence and experimental data for clinical treatment and further research.

Non-small cell lung cancer (NSCLC) remains a leading cause of cancer-related deaths globally. Current treatments often do not fully meet efficacy and quality of life expectations. Traditional Chinese medicine (TCM), particularly the Yiqi Sanjie formula, shows promise but lacks clear mechanistic understanding. This study addresses this gap by investigating the therapeutic effects and underlying mechanisms of Yiqi Sanjie formula in NSCLC. We utilized network pharmacology to identify potential NSCLC drug targets of the Yiqi Sanjie formula via the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Compounds with favorable oral bioavailability and drug-likeness scores were selected. Molecular docking was conducted using AutoDock Vina with structural data from the Protein Data Bank and PubChem. Molecular dynamics (MD) simulations were performed with Desmond Molecular Dynamics System, analyzing interactions up to 500 nanoseconds using the OPLS4 force field. ADMET predictions were executed using SwissADME and ADMETlab 2.0, assessing pharmacokinetic properties. Using network pharmacology tools, we performed Search Tool for the Retrieval of Interaction Genes/Proteins (STRING) analysis for protein-protein interaction, Kyoto Encyclopedia of Genes and Genomes (KEGG) for pathway enrichment, and gene ontology (GO) for functional enrichment, identifying crucial signaling pathways and biological processes influenced by the hit compounds bifendate, xambioona, and hederagenin. STRING analysis indicated substantial connectivity among the targets, suggesting significant interactions within the cell cycle regulation and growth factor signaling pathways as outlined in our KEGG results. The GO analysis highlighted their involvement in critical biological processes such as cell cycle control, apoptosis, and drug response. Molecular docking simulations quantified the binding efficiencies of the identified compounds with their targets-CCND1, CDK4, and EGFR-selected based on high docking scores that suggest strong potential interactions crucial for NSCLC inhibition. Subsequent MD simulations validated the stability of these complexes, supporting their potential as therapeutic interventions. Additionally, the novel identification of ADH1B as a target underscores its prospective significance in NSCLC therapy, further expanded by our comprehensive bioinformatics approach. Our research demonstrates the potential of integrating network pharmacology and computational biology to elucidate the mechanisms of the Yiqi Sanjie formula in NSCLC treatment. The identified compounds could lead to novel targeted therapies, especially for patients with overexpressed targets. The discovery of ADH1B as a therapeutic target adds a new dimension to NSCLC treatment strategies. Further studies, both in vitro and in vivo, are needed to confirm these computational findings and advance these compounds towards clinical trials.

Molecular and metabolic mechanisms of bufalin against lung adenocarcinoma: New and comprehensive evidences from network pharmacology, metabolomics and molecular biology experiment

Background and objectiveEpimedii has long been used as a traditional medicine in Asia for the treatment of various common diseases, including Alzheimer's disease, cancer, erectile dysfunction, and stroke. Studies have reported the ameliorative effects of Icariside II (ICS II), a major metabolite of Epimedii, on acute ischemic stroke (AIS) in animal models. Based on network pharmacology, molecular docking, and molecular dynamics (MD) simulations, we conducted a systematic review to evaluate the effects and neuroprotective mechanisms of ICS II on AIS.MethodsFirst, we have searched 6 databases using studies with ICS II treatment on AIS animal models to explore the efficacy of ICS II on AIS in preclinical studies. The literature retrieval time ended on March 8, 2022 (Systematic Review Registration ID: CRD42022306291). There were no restrictions on the language of the search strategy. Systematic review follows the Patient, Intervention, Comparison and Outcome (PICO) methodology and framework. SYCLE's RoB tool was used to evaluate the the risk of bias. In network pharmacology, AIS-related genes were identified and the target-pathway network was constructed. Then, these targets were used in the enrichments of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and gene ontology (GO). Molecular docking and MD simulation were finally employed between ICS II and the potential target genes.ResultsTwelve publications were included describing outcomes of 1993 animals. The literature details, animal strains, induction models, doses administered, duration of administration, and outcome measures were extracted from the 12 included studies. ICS II has a good protective effect against AIS. Most of the studies in this systematic review had the appropriate methodological quality, but some did not clearly state the controlling for bias of potential study. Network pharmacology identified 246 targets with SRC, CTNNB1, HSP90AA1, MAPK1, and RELA as the core target proteins. Besides, 215 potential pathways of ICS II were identified, such as PI3K-Akt, MAPK, and cGMP-PKG signaling pathway. GO enrichment analysis showed that ICS II was significantly enriched in subsequent regulation such as MAPK cascade. Molecular docking and MD simulations showed that ICS II can closely bind with important targets.ConclusionsICS II is a promising drug in the treatment of AIS. However, this systematic review reveals key knowledge gaps (i.e., the protective role of ICS II in women) that ICS II must address before it can be used for the treatment of human AIS. Our study shows that ICS II plays a protective role in AIS through multi-target and multi-pathway characteristics, providing ideas for the development of drugs for the treatment of AIS.