目的 運(yùn)用網(wǎng)絡(luò)藥理學(xué)的方法考察共軛亞油酸改善特應(yīng)性皮炎的作用機(jī)制，并通過動物進(jìn)行實驗驗證。方法 借助Swiss Target Prediction、STITCH數(shù)據(jù)庫獲取共軛亞油酸的作用靶點(diǎn)，DisGeNET、GeneCards、TTD數(shù)據(jù)庫檢索與特應(yīng)性皮炎相關(guān)的靶標(biāo)，利用Venny 2.1.0工具獲取共軛亞油酸與特應(yīng)性皮炎的交集靶點(diǎn)。采用STRING 11.0數(shù)據(jù)庫聯(lián)用Cytoscape 3.9.0軟件構(gòu)建交集靶點(diǎn)的蛋白相互作用（PPI）網(wǎng)絡(luò)，并篩選共軛亞油酸改善特應(yīng)性皮炎的核心靶點(diǎn)，并進(jìn)行基因本體（GO）功能富集分析和京都基因與基因組百科全書（KEGG）通路分析。將32只雌性昆明小鼠隨機(jī)分為對照組、模型組、共軛亞油酸（100 mg/kg）組和地塞米松（0.1 mg/kg）組，每組8只。采用局部涂抹2,4-二硝基氟苯（DNFB）建立特應(yīng)性皮炎小鼠模型，并比較各組小鼠皮損評分、皮膚組織病理學(xué)形態(tài)以及Th1/Th2型細(xì)胞因子水平。進(jìn)一步通過免疫組化檢測各組小鼠皮損中核心靶點(diǎn)PPARG的表達(dá)情況。結(jié)果 網(wǎng)絡(luò)數(shù)據(jù)庫共篩選出共軛亞油酸作用靶點(diǎn)108個，特應(yīng)性皮炎相關(guān)靶點(diǎn)基因1 708個，取交集后得到48個共軛亞油酸可能作用的特應(yīng)性皮炎靶點(diǎn)。核心靶點(diǎn)與KEGG通路分析結(jié)果顯示，共軛亞油酸主要作用于過氧化物酶體增殖物激活受體γ（PPARG）、脂聯(lián)素基因（ADIPOQ）等核心靶點(diǎn)及PPAR信號通路改善特應(yīng)性皮炎皮損癥狀。動物實驗結(jié)果發(fā)現(xiàn)，與模型組比較，共軛亞油酸組小鼠皮損評分顯著降低，炎癥細(xì)胞或肥大細(xì)胞的浸潤明顯減輕，Th1/Th2型細(xì)胞因子[免疫球蛋白E（IgE）、白細(xì)胞介素-4（IL-4）、γ干擾素（IFN-γ）]水平顯著降低（P＜0.05、0.01）。免疫組化檢測結(jié)果則發(fā)現(xiàn)，共軛亞油酸能顯著性上調(diào)特應(yīng)性皮炎小鼠皮損中PPARG的表達(dá)。結(jié)論 通過網(wǎng)絡(luò)藥理學(xué)和動物實驗初步驗證了共軛亞油酸對特應(yīng)性皮炎的改善作用及其可能的作用機(jī)制，為共軛亞油酸后續(xù)深入基礎(chǔ)實驗研究和臨床合理應(yīng)用提供科學(xué)依據(jù)。

Objective The mechanism of conjugated linoleic acid in improving atopic dermatitis was investigated by network pharmacology method and verified by animal experiments.Methods The targets of conjugated linoleic acid were obtained by Swiss Target Prediction and STITCH database.DisGeNET,GeneCards,and TTD databases were used to search for targets related to atopic dermatitis.The intersection targets of conjugated linoleic acid and atopic dermatitis were obtained using Venny 2.1.0 tool.The STRING 11.0 database and Cytoscape 3.9.0 software were used to construct the protein interaction (PPI) network of intersection targets.And screening the core target of conjugated linoleic acid to improve atopic dermatitis.Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed.Thirty-two female Kunming mice were randomly divided into control group,model group,conjugated linoleic acid (100 mg/kg) group and dexamethasone (0.1 mg/kg) group,with 8 mice in each group.The mouse model of atopic dermatitis was established by topical application of 2,4-dinitrofluorobenzene (DNFB),and the skin lesion score,skin histopathological morphology and Th1/Th2 cytokine levels were compared between the groups.The expression of core target PPARG in skin lesions was further detected by immunohistochemistry.Results A total of 108 target sites of conjugated linoleic acid and 1 708 target genes related to atopic dermatitis were screened from the network database,and 48 possible target sites of atopic dermatitis were obtained after intersection.The results of core target and KEGG pathway analysis showed that conjugated linoleic acid mainly acted on PPARG,ADIPOQ and other core targets,and PPAR signaling pathway to improve the symptoms of atopic dermatitis.The results of animal experiments showed that compared with the model group,the skin lesion score of mice in the conjugated linoleic acid group was significantly reduced,and the infiltration of inflammatory cells or mast cells was significantly reduced.The levels of Th1/Th2 cytokines[immunoglobulin E (IgE),interleukin-4(IL-4),interferon γ(IFN-γ)]were significantly decreased (P<0.05,0.01).The results of immunohistochemistry showed that conjugated linoleic acid could significantly up-regulate the expression of PPARG in skin lesions of mice with atopic dermatitis.Conclusion In this study,the ameliorative effect of conjugated linoleic acid on atopic dermatitis and its possible mechanism were initially verified through network pharmacology and animal experiments,which provided scientific basis for further basic experimental research and rational clinical application of conjugated linoleic acid.

R976