基于网络药理学与分子对接技术探讨利乳灵膳促进母乳分泌的潜在作用机制

Potential Mechanism of Liruling Diet in Promoting Breast Milk SecretionBased on Network Pharmacology and Molecular Docking Technology

目的:基于网络药理学和分子对接技术分析利乳灵膳促进母乳分泌的物质基础及潜在作用机制。方法:首先,通过BATMAN-TCM数据库及相关报道获取利乳灵膳组成药物的活性成分及作用靶点,检索GeneCards数据库获取与泌乳相关的靶点。其次,利用Venny 2.1在线分析平台获取共同靶点。然后,采用Cytoscape 3.7.2软件构建“药物-活性成分-共同靶点”网络图。利用STRING数据库及Cytoscape 3.7.2软件构建PPI图。通过HIPOLT绘图工具对共同靶点进行基因本体(gene ontology,GO)功能富集分析、京都基因组与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,利用AutoDock软件对利乳灵膳主要活性成分与核心靶点进行分子对接,以对GO功能富集分析、KEGG通路富集分析的调控网络进行靶向验证。结果:共得到共同靶点142个,核心靶点有信号转导和转录活化因子3(signal transducer and activator oftranscription 3,STAT3)、磷脂酰肌醇-4,5-二磷酸3-激酶催化亚基α(PIK3CA)、白细胞介素-6(interleukin-6,IL-6)等,所涉及的生物过程主要有肽分泌调节、分泌正性调节、蛋白质分泌调节等,细胞组成涉及线粒体基质、囊泡腔等,分子功能有信号受体激活剂活性、受体配体活性、激素活性等。机制主要涉及缺氧诱导因子-1(hypoxia inducible factor-1,HIF-1)、Janus激酶-信号转导子及转录激活子(janus kinase-sinal transducer and activator of transcription,JAK-STAT)信号通路等。各有效成分均能与缺乳、少乳核心靶点PIK3CA、STAT3(HIF-1、JAK-STAT关键靶点)、IL-6进行自发结合,并且白桦脂醇和延胡索乙素与PIK3CA和STAT3的结合能均小于阳性对照胃复安,具有较好的结合活性。结论:利乳灵膳可能通过核心靶点PIK3CA、STAT3、IL-6促进了STAT3磷酸化及HIF-1α表达而调控HIF-1、JAK-STAT信号通路,达到抑制乳腺细胞凋亡,促进多能干细胞自我更新、造血干细胞增殖分化、诱导HIF-1α表达而调控乳腺增殖、上调泌乳量的作用。

Objective:To analyze the material basis and potential mechanism of Liruling Diet in promoting breast milk secretion based on network pharmacology and molecular docking technology.Methods:Firstly,the active ingredients and targets of the drugs composed of Liruling Diet were obtained through the BATMAN-TCM database and related reports,and the targets related to lactation were obtained by searching the GeneCards database.Secondly,use the Venny 2.1 online analysis platform to obtain common targets.Then,the Cytoscape 3.7.2 software was used to construct the"drug-active ingredient-common target"network diagram.Using STRING database and Cytoscape 3.7.2 software to construct PPI diagram.Gene Ontology(GO)functional enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were carried out on the common targets by the HIPOLT drawing tool,and molecular docking of the main active ingredients and the core targets of Liruling Diet was carried out by the AutoDock software,so as to carry out target verification on the regulatory network of GO functional enrichment analysis and KEGG pathway enrichment analysis.Results:A total of 142 common targets were obtained,and the core targets included signal transducer and activator of transcription 3(STAT3),phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunitα(PIK3CA),interleukin-6(IL-6),etc.The biological processes involved mainly include the regulation of peptide secretion,positive regulation of secretion,regulation of protein secretion,etc.Cell composition involves mitochondrial matrix,vesicle cavity,etc.,and molecular functions include signal receptor activator activity,receptor-ligand activity,hormone activity,etc.The mechanism mainly involves hypoxia inducible factor-1(HIF-1),Janus kinase-sinal transducer and activator of transcription(JAK-STAT)signaling pathway,etc.Each active ingredient can spontaneously bind to PIK3CA,STAT3(HIF-1,JAK-STAT key targets),and IL-6,the core targets of agalactia and hypogalactia.Moreover,the binding energies of betulin and tetrahydropalmatine to PIK3CA and STAT3 were both lower than those of the positive control metoclopramide,which shows good binding activity.Conclusion:Liruling diet may promote the phosphorylation of STAT3 and the expression of HIF-1αthrough the core targets PIK3CA,STAT3,and IL-6,and regulate the HIF-1 and JAK-STAT signaling pathways,thus to inhibit the apoptosis of mammary gland cells,promote the self-renewal of pluripotent stem cells,promote the proliferation and differentiation of hematopoietic stem cells,and induce the expression of HIF-1α,so as to regulate the proliferation of mammary glands,and increase the milk production.