基于网络药理学与实验验证探讨藏药五味甘露药浴组方治疗银屑病的作用机制OA

Objective To investigate the key targets and pathways of the classic Tibetan medicine formula Wuwei Ganlu Medicated Bath in the treatment of psoriasis using network pharmacology and molecular docking techniques,and to validate the relevant findings through cellular experiments.Methods Essential oils from the constituent herbs of the Wuwei Ganlu Medicated Bath formula—Vkhan Pa,Mtshe Ldum,Shug Pa,Da Lis,and Vom Bu—were extracted using supercritical CO2 extraction and identified by gas chromatography-mass spectrometry(GC-MS).Key components and their targets were screened using databases such as SwissTargetPrediction.Psoriasis-related targets were obtained from databases including the Online Mendelian Inheritance in Man(OMIM).After identifying intersecting targets,a protein-protein interaction(PPI)network was constructed,followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.Molecular docking validation between core components and key targets was performed using software such as Autodock.Finally,in vitro cellular experiments were conducted to validate the predictions.A psoriasis inflammation model was established using lipopolysaccharide(LPS)-induced human adult low calcium high temperature(HaCaT)keratinocytes.Safe administration concentrations were determined using the Cell Counting Kit-8(CCK-8)assay.The regulatory effects of totarol,trans-nerolidol,lobetyolin acetate,and cryptomeridiol on the inflammatory factors interleukin-17(IL-17),interleukin-23(IL-23),interleukin-6(IL-6),and interferon-γ(IFN-γ)were detected by enzyme-linked immunosorbent assay(ELISA).Results From the 174 compounds identified by GC-MS,42 key components were selected based on a skin permeability coefficient(log Kp)≥-6 cm·s-1.A total of 109 intersecting drug-disease targets were obtained.PPI network and drug-component-disease-target network analyses identified 7 core targets,including signal transducer and activator of transcription 3(STAT3),Janus kinase 2(JAK2),Janus kinase 1(JAK1),B-cell lymphoma 2(BCL2),and epidermal growth factor receptor(EGFR).These interacted with core components such as totarol,trans-nerolidol,lobetyolin acetate,cryptomeridiol,and(4-phenyl)but-2-yl acetate,primarily involving pathways related to cell proliferation,apoptosis,immune regulation,and inflammation.Molecular docking indicated strong binding affinity of totarol with EGFR,JAK1,and BCL2.Cellular experiments confirmed that all components were non-cytotoxic at 20 µg·mL-1 and significantly inhibited the secretion of IL-17,IL-23,and IFN-γ(P＜0.01).Regarding IL-6 inhibition,lobetyolin acetate and cryptomeridiol showed significant effects(P＜0.01),and totarol also exhibited a notable effect(P＜0.05),confirming that these components can inhibit the expression of psoriasis-related inflammatory factors through multiple targets.Conclusion The Tibetan medicine Wuwei Ganlu Medicated Bath formula is a clinically common external treatment for psoriasis in Tibetan medicine due to its favorable efficacy.Through multi-dimensional"component-target-pathway"analysis and experimental validation,this study found that its core active components exert anti-inflammatory and immunomodulatory effects by targeting key targets such as STAT3 and JAK1,primarily via the JAK-STAT signaling pathway,inhibiting the expression of characteristic inflammatory factors including IL-17,IL-23,and IFN-γ.This elucidates the specific molecular mechanism underlying its therapeutic effect via the"Yellow Water(a pathological body fluid in Tibetan medicine)-skin-psoriasis"pathway from a modern pharmacological perspective,providing a basis for revealing the scientific connotation of the Wuwei Ganlu Medicated Bath formula in treating skin diseases.