白鲜皮免疫调节机制的网络药理学分析及验证

Network Pharmacology Analysis and Verification of ImmunomodulatoryMechanism of Cortex Dictamni

本试验旨在探究白鲜皮的免疫调节能力及其机制。通过TCMSP数据库筛选出11个白鲜皮活性成分,利用GeneCard数据库和OMIM数据库预测到318个相关联的靶点基因,通过对药物靶点的筛选,获得63个疾病与药物共同的靶点。通过Cytoscape软件构建药物-活性成分-靶点网络。使用Perl计算机程序语言对GO功能富集结果进行筛选和KEGG通路富集分析。通过腹腔注射环磷酰胺建立免疫抑制小鼠模型,用1.3、2.6、5.2 g/kg BW(按生药量计)的白鲜皮提取物灌胃免疫抑制小鼠2周后检测小鼠体重、免疫器官指数、血清细胞因子含量、血常规指标。结果显示:白鲜皮中的黄芩素、β-谷甾醇、木犀草素、白鲜明碱、前茵芋碱、槲皮素等化合物通过调节AKT1、IL6、IL1B、JUN、PTGS2、VEGFA等重要靶点,参与调节白细胞介素-17信号通路、肿瘤坏死因子信号通路,进而起到免疫调节的效果。小鼠免疫抑制试验发现,相比模型组,5.2 g/kg BW白鲜皮提取物对小鼠的体重没有产生显著影响(P>0.05),但极显著增加了小鼠的肝脏指数和脾脏指数(P<0.01);此外,5.2 g/kg BW白鲜皮提取物极显著提高了小鼠血清白细胞介素-2(IL-2)、干扰素-γ(IFN-γ)、免疫球蛋白A(IgA)和肿瘤坏死因子-α(TNF-α)含量(P<0.01)。本研究展现了白鲜皮免疫调节多成分、多靶点作用的特点,可为白鲜皮在畜禽健康养殖中的应用提供科学依据。

This experiment was conducted to investigate the immunomodulatory ability and its mechanism of Cortex Dictamni.Eleven active ingredients of Cortex Dictamni were screened out from the TCMSP database,318 related target genes were predicted from the GeneCard database and OMIM database,and 63 common targets of diseases and drugs were obtained through screening with drug targets.Cytoscape software was used to construct the“drug-active ingredients-target”network.GO functional enrichment analysis results were screened and KEGG pathway enrichment analysis was performed using the Perl computer programming language.The immunosuppressed mouse model was established by intraperitoneal injection of cyclophosphamide,and the immunosuppressed mice were treated with different doses(1.3,2.6 and 5.2 g/kg BW,calculated by the amount of crude drug)of extract of Cortex Dictamni by gavage for 2 weeks,and then the body weight,immune organ indexes,serum cytokine contents,and blood routine indexes of mice were detected.The results showed that baicalein,β-sitosterol,luteolin,alpinine,proalpinine,quercetin and other major compounds in Cortex Dictamni were involved in interleukin-17(IL-17)and tumor necrosis factor(TNF)signaling pathways by regulating key targets such as AKT1,IL6,IL1B,JUN,PTGS2,VEGFA and so on.The immunosuppression test showed that compared with the model group,the 5.2 g/kg BW of Cortex Dictamni extract had no significant effects on the weight gain of mice(P>0.05),but had extremely significant effects on the liver and spleen indexes(P<0.01);in addition,the 5.2 g/kg BW of Cortex Dictamni extract could extremely significantly increase the contents of serum interleukin-2(IL-2),interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α)and immunoglobulin A(IgA)of mice(P<0.01).This study shows the characteristics of multi-component and multi-target immune regulation of Cortex Dictamni,which provides a scientific basis for the application of Cortex Dictamni in healthy livestock and poultry breeding.