目的:基于网络药理学和动物实验探讨桑菊饮治疗急性肺损伤(acute lung injury,ALI)的作用机制.方法:采用中药系统药理学数据库与分析平台检索桑菊饮的化学成分,并通过《中华人民共和国药典》加以补充.利用Swiss Target Prediction数据库获取桑菊饮的靶点,GeneCards数据库获取ALI的靶点,并借助Venny平台得到两者的交集靶点.通过Cytoscape软件构建"疾病-中药-活性成分-靶点"网络,经过拓扑分析筛选桑菊饮治疗ALI的关键成分.采用STRING数据库构建蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,并筛选桑菊饮治疗ALI的核心靶点.通过Metascape网站进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)分析,最后进行分子对接验证.18 只Wistar大鼠随机分为空白组、模型组、桑菊饮组(4g·kg-1),每组6 只.桑菊饮组大鼠灌胃相应药物,空白组和模型组大鼠给予等体积生理盐水,连续5d.末次灌胃后2h,除空白组外,其他大鼠经气管插管注射脂多糖(5mg·kg-1)以复制ALI模型.采用ELISA法检测大鼠肺组织白细胞介素-6(interleukin-6,IL-6)、丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量.结果:桑菊饮活性成分171 个,作用靶点505 个.ALI疾病靶点8 053 个,两者的交集靶点447 个.桑菊饮治疗ALI的关键成分主要包括木犀草素、槲皮素、桦木酸等.桑菊饮治疗ALI的核心靶点主要包括RAC-α丝氨酸/苏氨酸蛋白激酶(RAC-alpha serine/threonine protein ki-nase,AKT1)、信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)、细胞凋亡调节剂Bcl-2(ap-optosis regulator Bcl-2,BCL2)等.GO分析结果主要涉及磷酸化、激素反应、氧化还原酶活性等.KEGG通路主要包括Th17细胞分化、JAK-STAT信号通路、AMPK信号通路等.分子对接结果显示,关键成分与核心靶点具有较强的结合活性.ELISA结果显示,与空白组比较,模型组大鼠肺组织IL-6、MDA、TNF-α含量升高(P<0.01),SOD含量降低(P<0.01);与模型组比较,桑菊饮组大鼠肺组织IL-6、MDA、TNF-α含量降低(P<0.01),SOD含量升高(P<0.01).结论:桑菊饮可通过多成分、多通路、多靶点发挥治疗ALI的作用,其作用机制可能与减轻炎症反应与氧化损伤有关.
Study on the Mechanism of Mulberry Leaf and Chrysanthemum Beverage in Treating Acute Lung Injury Based on Network Pharmacology and Animal Experiments
Objective:To study the mechanism of Mulberry Leaf and Chrysanthemum Beverage in treating acute lung injury(ALI)based on network pharmacology and animal experiments.Methods:The chemical components of Mulberry Leaf and Chrysanthemum Beverage were retrieved using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and supple-mented by The Pharmacopoeia of the People's Republic of China.Swiss Target Prediction database was used to obtain the targets of Mulberry Leaf and Chrysanthemum Beverage,and GeneCards database was used to obtain the targets of ALI,and Venny platform was used to obtain the intersection targets of the two.The network of"disease-Chinese medicinal-active ingredient-target"was constructed using Cytoscape software,and the key components of Mulberry Leaf and Chrysanthemum Beverage for the treatment of ALI were screened through the topology analysis.The protein-protein interaction(PPI)network was constructed using the STRING database and the core targets of Mulberry Leaf and Chrysanthemum Beverage in treating ALI were screened.Gene Ontology(GO)and Kyoto Encyclo-pedia of Genes and Genomes(KEGG)analysis was performed through the Metascape website,and finally perform molecular docking validation was performed.A total of 18 Wistar rats were randomly divided into the blank group,the model group,and the group of Mul-berry Leaf and Chrysanthemum Beverage(4 g·kg-1),with6 rats in each group.The rats in the group of Mulberry Leaf and Chrysan-themum Beverage were given corresponding drugs by gavage,while the blank group and model group rats were given equal volumes of physiological saline for 5 consecutive days.And 2 hours after the last gavage,except the blank group,other rats were injected with li-popolysaccharide(5 mg·kg-1)through tracheal intubation to replicate the ALI model.ELISA was used to detect the contents of in-terleukin-6(IL-6),malondialdehyde(MDA),superoxide dismutase(SOD),and tumor necrosis factor-α(TNF-α)in rat lung tissue.Results:There were 171 active components and 505 targets of action in Mulberry Leaf and Chrysanthemum Beverage.There were 8 053 ALI disease targets,with 447 intersecting targets.The key components of Mulberry Leaf and Chrysanthemum Beverage in treating ALI mainly included luteolin,quercetin,and mairin,etc.The core targets of Mulberry Leaf and Chrysanthemum Beverage in treating ALI mainly included RAC-alpha serine/threonine protein kinase(AKT1),signal transducer and activator of transcription 3(STAT3),and apoptosis regulator Bcl-2(BCL2),etc.The GO analysis results mainly involved phosphorylation,hormone reactions,oxidoreductase ac-tivity,etc.The KEGG pathway mainly included Th17 cell differentiation,JAK-STAT signaling pathway,AMPK signaling pathway,etc.The molecular docking results showed that key components(such as luteolin)have strong binding activity with core targets(such as AKT1).The ELISA results showed that compared with the blank group,the contents of IL-6,MDA,TNF-α in the lung tissue of the model group rats increased(P<0.01),while the content of SOD decreased(P<0.01);compared with the model group,the contents of IL-6,MDA,and TNF-α in the lung tissue of rats in the group of Mulberry Leaf and Chrysanthemum Beverage decreased(P<0.01),while the content of SOD increased(P<0.01).Conclusion:Mulberry Leaf and Chrysanthemum Beverage can exert therapeutic effects on ALI through multiple components,pathways,and targets,and its mechanism may be related to reducing inflammatory response and oxidative damage.
Mulberry Leaf and Chrysanthemum Beverageacute lung injury(ALI)network pharmacologymolecular dockingluteolinrats
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