摘要
目的:通过体内实验初步验证白鲜皮治疗湿疹的有效性,并通过网络药理学及实验验证探讨白鲜皮治疗湿疹的主要活性成分及其潜在作用机制。方法:30只雌性Balb/c小鼠,随机分为空白对照组、模型组、白鲜皮组,每组各10只。除空白对照组外,其余各组采用2,4-二硝基氯苯(DNCB)涂于小鼠腹部诱导致敏反应,建立湿疹模型。白鲜皮组小鼠灌胃给予白鲜皮水提液0.5 g生药/(kg·d),空白对照组和模型组经灌胃给予等体积0.9%NaCl溶液。采用中药系统药理分析平台(TCMSP)检索白鲜皮活性成分并获取相应靶点,在GeneCards数据库中检索湿疹靶点,运用Cytoscape软件构建“白鲜皮-活性成分-潜在靶标”网络,通过STRING数据库构建蛋白-蛋白相互作用(PPI)网络,将潜在靶点导入DAVID数据库进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析。采用酶联免疫吸附试验(ELISA)检测各组小鼠血清中白介素(IL)-6、IL-10和肿瘤坏死因子-α(TNF-α)水平,验证网络药理学分析结果。结果:体内试验结果显示,白鲜皮能缓解湿疹造成的小鼠耳片肿胀,明显降低小鼠耳片肿胀率(P<0.01)。网络药理学分析显示,白鲜皮中的主要活性成分为白鲜碱、槲皮素、黄柏酮、汉黄芩素、甘遂烷、β-谷甾醇、前茵芋碱。白鲜皮作用于湿疹的潜在靶点有44个,关键靶标主要包括TNF-α、IL-1β、IL-6等。富集分析显示,白鲜皮治疗湿疹涉及的生物过程和信号通路主要有脂多糖应答、炎症反应、IL-17信号通路、细胞凋亡信号通路等。ELISA检测结果显示,与对照组比较,模型组小鼠血清中IL-6、IL-1β、TNF-α水平明显升高(P<0.01);与模型组比较,白鲜皮组小鼠血清中IL-6、IL-1β、TNF-α水平明显降低(P<0.01)。结论:白鲜皮可能通过多成分、多靶点、多通路的方式治疗湿疹,并能够降低相关炎症因子(IL-6、IL-10、TNF-α)水平。
Objective:To preliminarily verify the efficacy of Dictamni Cortex in the treatment of eczema in vivo,and to explore the main active ingredients and potential mechanisms of Dictamni Cortex in the treatment of eczema through network pharmacology and experimental verification.Methods:Totally 30 healthy female Balb/c mice were randomly divided into blank control group,model group,Dictamni Cortex group,with 10 mice in each group.Except the blank control group,the other groups were sensitized with 2,4‐dinitrochlorobenzene(DNCB)on the abdominal skin of mice to establish eczema model.The mice in the Dictamni Cortex group were given 0.5 g crude drug/(kg·d)Dictamni Cortex water extract by gavage,while the blank control group and the eczema model group were given the same volume of 0.9%NaCl solution by gavage.Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)was used to retrieve components and collect the corresponding targets.GeneCards database was used to search for disease targets.Cytoscape software was used to construct"Dictamni Cortex-ingredients-potential targets"network.The protein-protein interaction(PPI)network was built through STRING database,and the potential targets were imported into the DAVID database for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses.The serum levels of interleukin(IL)-6,IL-10 and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA)to verify the results of network pharmacology analysis.Results:The results of in vivo experiments showed that the Dictamni Cortex can alleviate the swelling of mice ears caused by eczema and significantly reduce the swelling rate of mice ears(P<0.01).The network pharmacology analysis showed that the key active ingredients in Dictamni Cortex were dasycarpamin,quercetin,obacunone,wogonin,tirucallane,β‐sitosterol,preskimmianine.There were 44 potential targets of leukoderma on eczema,and the key targets mainly include TNF-α,IL-1β,IL-6,etc.Enrichment analyses showed that the biological processes and signaling pathways involved in the treatment of eczema by Dictamni Cortex mainly included lipopolysaccharide response,inflammatory response,IL-17 pathway,and apoptotic signaling pathway,etc.ELISA results showed that compared with the control group,the serum levels of IL-6,IL-1β,and TNF-αin the model group were significantly increased(P<0.01).Compared with the model group,the serum levels of IL-6,IL-1βand TNF-αin the Dictamni Cortex group were significantly decreased(P<0.01).Conclusion:Dictamni Cortex may treat eczema through a multi-component,multi-target,and multi‐pathway approach,and can reduce the levels of related inflammatory factors(IL-6,IL-10 and TNF-α).
作者
卢懿
吴伟
杨扬
张彤
刘梅
LU Yi;WU Wei;YANG Yang;ZHANG Tong;LIU Mei(Shanghai Innovation Center of TCM Health Service,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Tufeng Pharmaceutical Technology Company Limited,Shanghai 201203,China;Science and Technology Experimental Center,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;School of Pharmacy,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)
出处
《上海中医药大学学报》
CAS
2024年第5期62-70,共9页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
康缘医药研究院新药孵化基金项目(20210906)。
关键词
白鲜皮
湿疹
体内实验
网络药理学
靶点
炎症因子
Dictamni Cortex
eczema
in vivo experiment
network pharmacology
target
inflammatory factors
