白鲜皮及汉黄芩素治疗银屑病的网络药理学研究及细胞验证  

Network Pharmacology Study and Cellular Validation of Dictamni Cortex and Wogonin in the Treatment of Psoriasis

作　　者：林丽云[1] 林钰 雷海青 刘靖[2] 张粤琳 黄伟乐 杨丽红[2] 陶佳豪 LIN Li-Yun;LIN Yu;LEI Hai-Qing;LIU Jing;ZHANG Yue-Lin;HUANG Wei-Le;YANG Li-Hong;TAO Jia-Hao(Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Dept.of Dermatology,The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学,广东广州510405 [2]广州中医药大学第一附属医院皮肤科,广东广州510405

出　　处：《广州中医药大学学报》2023年第6期1488-1497,共10页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：广东省教育厅普通高校重点领域专项(编号:2021ZDZX2026);广东省中医药局资助课题项目(编号:20213006);国家自然科学基金青年科学基金资助课题项目(编号:812026990)。

摘　　要：【目的】通过网络药理学探讨中药白鲜皮及其主要活性成分汉黄芩素治疗银屑病靶点和机制,并进行细胞验证。【方法】通过中药系统药理学数据库与分析平台(TCMSP)、中医百科全书数据库检索白鲜皮化学成分,以口服生物利用度(OB)和类药性(DL)筛选其主要活性成分及作用靶点,在GeneCard数据库检索银屑病疾病靶点,对二者取交集得到白鲜皮潜在调控靶点。对这部分靶点进行基因富集分析,并使用STRING数据库、Cytoscape软件制作蛋白互作图,再进一步使用CytoHubba插件进行关键基因的筛选。选取调控关键基因最多的主要活性成分汉黄芩素进行细胞实验:制作HaCaT细胞银屑病样模型,确定汉黄芩素的半抑制浓度(IC50),以10%IC50干预银屑病样细胞模型,对关键基因进行实时定量聚合酶链反应(qPCR)检测。【结果】白鲜皮共筛选出11种有效单体成分,共计对应106个有效靶点,银屑病包含4055个可供调控的靶点,取交集发现其中63个可能是白鲜皮治疗银屑病的潜在调控靶点。基因富集分析结果显示白鲜皮主要有效成分调控银屑病主要通过免疫与炎症相关通路。CytoHubba插件筛选出10个关键基因,主要受槲皮素、木犀草素、汉黄芩素调控。汉黄芩素进一步被作为研究对象,处理以1μg/mL脂多糖(LPS)诱导细胞4 h构建的HaCaT细胞银屑病样模型,汉黄芩素对HaCaT细胞的IC50为415μmol/L。以汉黄芩素10%IC50干预模型细胞,结果显示其可以显著提高PPARG mRNA表达,降低ESR1、HSP90AA1、IL-6、NF-κB、PTGS2、TNF-α、TP53 mRNA表达(P<0.005)。【结论】白鲜皮主要有效成分汉黄芩素可能通过调控TNF-α、IL-6、NF-κB、TP53、HSP90AA1、ESR1、PTGS2、PPARG mRNA表达,降低炎性水平,从而治疗银屑病。Objective To explore the mechanism and the therapeutic targets of the Chinese herbal medicine Dictamni Cortex and the main active ingredient wogonin for psoriasis through network pharmacology and cellular validation.Methods The chemical composition of Dictamni Cortex was searched through the Systematic Pharmacology Database and Analysis Platform of Traditional Chinese Medicine(TCMSP)and the Encyclopedia of Chinese Medicine database,and the main active ingredients and targets were screened by oral bioavailability(OB)and drug-like properties(DL),the GeneCard database was searched for psoriasis disease targets,and the two were intersected to obtain the potential regulatory targets of Dictamni Cortex.Gene enrichment analysis was performed on this part of the targets,and the STRING database and Cytoscape software were used to produce interactions,and further key genes were screened using the CytoHubba plugin.Cellular experiments were performed on the key active ingredients that regulate the most key genes:HaCaT cell psoriasis-like model was produced,semi-inhibitory concentrations(IC50)of the key active ingredients was determined,10%IC50 was used in intervening psoriasis-like cell model and real-time quantitative polymerase chain reaction(qPCR)assay was performed for key gene changes.Results A total of 11 active monomeric components of Dictamni Cortex were screened,corresponding to a total of 106 active targets,psoriasis contained 4055 targets for regulation,of which 63 were found to be potential regulatory targets for the treatment of psoriasis with Dictamni Cortex after the intersection.The results of the gene enrichment analysis suggested that the main active components of Dictamni Cortex regulated psoriasis mainly through immune and inflammatory related pathways.The CytoHubba plug-in screened 10 key genes,mainly regulated by quercetin,lignocaine and wogonin.Wogonin was used as a further study to treat HaCaT cell psoriasis-like model induced by 1μg/mL lipopolysaccharide(LPS)for 4 h.The IC50 of wogonin against HaCaT

关 键 词：白鲜皮 银屑病 汉黄芩素 网络药理学 炎症通路 免疫通路 HACAT细胞 

分 类 号：R285.5[医药卫生—中药学]