【摘要】 目的：基于超高效液相色谱-四级杆飞行时间质谱(UPLC-Q-TOF-MS/MS)技术和网络药理学探讨柔肝方治疗肝纤维化的作用机制。方法：将8只C57BL/J小鼠随机分成空白血清组和柔肝方含药血清组，每组4只。利用UPLC-Q-TOF-MS/MS技术结合Xcalibur软件分析空白血清和柔肝方含药血清数据，结合二级谱图鉴定柔肝方的入血成分。通过Swiss Target Prediction数据库和中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform, TCMSP)检索柔肝方入血成分的靶点。利用GeneCards、OMIM、PharmGKB、DrugBank Online和Therapeutic Target Database数据库检索肝纤维化的靶点。采用Cytoscape软件构建“疾病-化合物-靶点”网络，并筛选柔肝方抗肝纤维化的关键成分。借助STRING数据库构建蛋白质-蛋白质相互作用(protein-protein interaction, PPI)网络，通过拓扑分析筛选柔肝方抗肝纤维化的核心靶点。采用DAVID数据库进行基因本体(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。结果：柔肝方入血成分主要包括氧化槐果碱、槐果碱、高黄芩素、新补骨脂异黄酮等。通过检索获取柔肝方入血成分相关靶点303个，肝纤维化相关靶点6 331个，两者的交集靶点250个。柔肝方抗肝纤维化的关键成分包括槲皮素、芹菜素、山柰酚、芒柄花黄素等。柔肝方抗肝纤维化的核心靶点包括原癌基因酪氨酸蛋白激酶Src(SRC)、信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)、丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、丝氨酸/苏氨酸蛋白激酶(protein kinase B,PKB,又称AKT1)等。GO分析主要涉及细胞因子介导的信号通路、凋亡过程的负调控、炎症反应、蛋白激酶活性等。KEGG分析主要涉及PI3K-Akt信号通路、MAPK信号通路、白细胞介素(interleukin, IL)-17信号通路、HIF-1信号通路等。结论：柔肝方可能通过槲皮素、芹菜素、山柰酚等化合物调控PI3K-Akt、MAPK、IL-17等信号通路，作用于SRC、STAT3、MAPK1等靶点，从而发挥抗肝纤维化的作用，体现了多成分、多途径、多靶点的药理特征。更多还原

【Abstract】 Objective: To study the therapeutic mechanism of Liver-Softening Formula in treating liver fibrosis based on ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) technology and network pharmacology.Methods: A total of 80 C57BL/J mice were randomly divided into the blank serum group and the group of drug-containing serum from Liver-Softening Formula, with 4 mice in each group.The data of blank serum and drug-containing serum from Liver-Softening Formula were analyzed by UPLC-Q-TOF-MS/MS technology combined with Xcalibur software, and the blood-entering components of Liver-Softening Formula were identified combined with secondary mass spectrum.By using Swiss Target Prediction database and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),the targets of blood-entering components in Liver-Softening Formula were searched.The targets of liver fibrosis were searched using GeneCards, OMIM,PharmGKB,DrugBank Online, and Therapeutic Target Database databases.Using Cytoscape software, a "disease-compound-target" network was constructed, and the key components of Liver-Softening Formula in its anti-liver fibrosis effect were screened.The protein-protein interaction(PPI) network was constructed with the help of STRING database, and the core targets of Liver-Softening Formula in its anti-liver fibrosis were screened by topological analysis.Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) were analyzed using the DAVID database.Results: The main components of Ruogan prescription into blood included Oxysophocarpine, Sophocarpine, Scutellarein, Neobavaisoflavone, etc.A total of 303 related targets and 6 331 related targets of liver fibrosis, and 250 intersection targets of the two were obtained.The key components of Liver-Softening Formula in its anti-liver fibrosis effect include Quercetin, Apigenin, Kaempferol, Formononetin and so on.The core targets of Liver-Softening Formula in its anti-liver fibrosis effect included proto-oncogene tyrosine protein kinase Src(SRC),signal transducer and activator of transcription 3(STAT3),mitogen activated protein kinase 1(MAPK1),serine/threonine protein kinase B(PKB,also known as AKT1),etc.The GO analysis mainly involved the Cytokine-mediated signaling pathway and the negative regulation of apoptotic process process, inflammatory response, protein kinase activity, etc.KEGG analysis mainly involved PI3K-Akt signaling pathway, MAPK signaling pathway, IL-17 signaling pathway, HIF-1 signaling pathway, etc.Conclusion: Liver-Softening Formula may regulate PI3K-Akt, MAPK,IL-17 and other signaling pathways through quercetin, apigenin, kaemphiol and other compounds, and act on SRC,STAT3,MAPK1 and other targets, thus playing an anti-fibrosis role, reflecting the pharmacological characteristics of multi-components, multi-pathways and multi-targets.更多还原