目的 运用网络药理学的方法考察共轭亚油酸改善特应性皮炎的作用机制，并通过动物进行实验验证。方法 借助Swiss Target Prediction、STITCH数据库获取共轭亚油酸的作用靶点，DisGeNET、GeneCards、TTD数据库检索与特应性皮炎相关的靶标，利用Venny 2.1.0工具获取共轭亚油酸与特应性皮炎的交集靶点。采用STRING 11.0数据库联用Cytoscape 3.9.0软件构建交集靶点的蛋白相互作用（PPI）网络，并筛选共轭亚油酸改善特应性皮炎的核心靶点，并进行基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路分析。将32只雌性昆明小鼠随机分为对照组、模型组、共轭亚油酸（100 mg/kg）组和地塞米松（0.1 mg/kg）组，每组8只。采用局部涂抹2,4-二硝基氟苯（DNFB）建立特应性皮炎小鼠模型，并比较各组小鼠皮损评分、皮肤组织病理学形态以及Th1/Th2型细胞因子水平。进一步通过免疫组化检测各组小鼠皮损中核心靶点PPARG的表达情况。结果 网络数据库共筛选出共轭亚油酸作用靶点108个，特应性皮炎相关靶点基因1 708个，取交集后得到48个共轭亚油酸可能作用的特应性皮炎靶点。核心靶点与KEGG通路分析结果显示，共轭亚油酸主要作用于过氧化物酶体增殖物激活受体γ（PPARG）、脂联素基因（ADIPOQ）等核心靶点及PPAR信号通路改善特应性皮炎皮损症状。动物实验结果发现，与模型组比较，共轭亚油酸组小鼠皮损评分显著降低，炎症细胞或肥大细胞的浸润明显减轻，Th1/Th2型细胞因子[免疫球蛋白E（IgE）、白细胞介素-4（IL-4）、γ干扰素（IFN-γ）]水平显著降低（P＜0.05、0.01）。免疫组化检测结果则发现，共轭亚油酸能显著性上调特应性皮炎小鼠皮损中PPARG的表达。结论 通过网络药理学和动物实验初步验证了共轭亚油酸对特应性皮炎的改善作用及其可能的作用机制，为共轭亚油酸后续深入基础实验研究和临床合理应用提供科学依据。

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