黄白双花口服液治疗犊牛腹泻的系统药理学分析

【目的】利用系统药理学构建黄白双花口服液治疗犊牛腹泻的活性化合物—靶点—通路—疾病相关蛋白的关系网络模型，阐述黄白双花口服液治疗犊牛腹泻的分子机制，为犊牛腹泻防治药物研发提供理论依据。【方法】根据口服利用度（OB）≥30%和类药性（DL）≥0.18，利用中药系统药理学数据库（TCMSP）检索黄白双花口服液中每味中药（黄连、白头翁、金银花、黄芩、地榆和乌梅）的活性化合物及与其相对应的作用靶点，导入Cytoscape绘图软件中建立活性化合物与其对应靶点的互作网络图；通过基因数据库（Gene Cards）检索犊牛腹泻的有关靶点基因，使用维恩图求取二者的交集，得到犊牛腹泻与黄白双花口服液共同的靶点基因组合，然后导入String数据库中以获得相关蛋白相互作用网络图；将黄白双花口服液作用于犊牛腹泻的靶点基因导入DAVID数据库中，分别进行GO功能富集分析和KEGG通路富集分析。【结果】从黄白双花口服液中筛选出43种活性化合物，分别对应131个相关靶点基因。黄白双花口服液与犊牛腹泻交集的蛋白质—蛋白质相互作用网络图包括128个节点及205条相互关系，其中丝氨酸/苏氨酸蛋白激酶1（AKT1）、白介素2（IL-2）、白介素6（IL-6）、白介素8（IL-8）、促分裂素原活化蛋白激酶1（MAPK1）及含半胱氨酸的天冬氨酸蛋白酶3（CASP3）的出现频率较高。利用DAVID数据库对黄白双花口服液与犊牛腹泻交集靶点的蛋白进行GO功能富集分析，最终富集到204条GO功能条目（FDR< 10-2），其中细胞组成条目8条、分子功能条目12条、生物过程条目184条；黄白双花口服液治疗犊牛腹泻的相关蛋白主要富集于19条信号通路上（FDR< 10-2）。【结论】黄白双花口服液主要以黄连、白头翁、金银花、黄芩、地榆和乌梅中的槲皮素、β-谷甾醇、豆甾醇及山奈酚等活性化合物成分，通过ATK1、MAPK1、IL-2、IL-6和IL-8等靶点，经Toll样受体信号通路、NOD样受体信号通路、T细胞受体信号通路和B细胞受体信号通路等，发挥抗菌消炎、抑制细胞损伤及调节免疫力等作用，以达到治疗犊牛腹泻的目的。

A systematic pharmacological analysis of Huangbaishuanghua oral liquid for treating calf diarrhea

【Objective】Based on systematic pharmacology, a network of active compound, target, pathway and disease-related protein of Huangbaishuanghua oral liquid was established to elucidate the molecular mechanism of treatingdiarrhea in calves, and to provide the theoretical basis for research and development of drugs for the prevention and treatment of diarrhea in calves.【Method】According to oral utilization(OB) ≥ 30% and drug-likeness(DL) ≥ 0.18, using traditional Chinese medicine pharmacology(TCMSP) database retrieval and double liquid medium every taste traditional Chinese medicine(Coptidis Rhizoma, Pulsatilliae Radix, Lonicerae Japonicae Flos, Scutellariae Radix, Radix Sanguisorbae and Mume Fructus) of the active compounds and corresponding targets, imported into Cytoscape drawing software to establish active compounds and their corresponding targets interactions network diagram.The target genes of calf diarrhea were retrieved through Gene Cards database, and the intersection of the two was obtained by Venn diagram.The target gene combination of calf diarrhea and Huangbaishuanghua oral liquid was obtained, and then imported into the String database to obtain the network diagram of the interaction of related proteins.The target genes of Huangbaishuanghua oral liquid on diarrhea of calves were imported into DAVID database for GO function enrichment analysis and KEGG pathway enrichment analysis, respectively.【Result】Forty-three active compounds were screened from Huangbaishuanghua oral Liquid, corresponding to 131 related target genes, respectively.Huangbai Shuanghua Oral Liquid and calf diarrhea intersection protein-protein intera-ction network diagram includded 128 nodes and 205 relationship, among them, serine/threonine protein kinase 1(AKT1), interleukin 2(IL-2) and interleukin 6(IL-6) and interleukin 8(IL-8), promote the mitogen activated protein kinase 1(MAPK1) and aspartic acid with cysteine proteinase 3(CASP3) had high occurrence frequency.The GO function enrichment analysis of the proteins at the intersection target of Huangbaishuanghua oral liquid and diarrhea of calfs was conducted using DAVID database.Finally, 204 GO functional items(FDR< 10-2) were enriched, including 8 cell composition items, 12 molecular function items and 184 biological process items.The related proteins of Huangbaishuanghua oral liquid in the treatment of diarrhea in calves were mainly enriched in 19 signaling pathways(FDR< 10-2).【Conclusion】Huangbaishuanghua oral liquid mainly use quercetin, β-phytosterol, stigstanol and kaempferol from Coptidis Rhizoma, Pulsatilliae Radix, Lonicerae Japonicae Flos, Scutellariae Radix, Radix Sanguisorbae and Mume Fructus.Through ATK1, MAPK1, IL-2, IL-6 and IL-8 and other targets, it can play antibacterial and anti-inflammatory roles, inhibit cell damage and regulate immunity through Toll-like receptor signaling pathway, NOD receptor signaling pathway, T cell receptor signaling pathway and B cell receptor signaling pathway, to achieve the purpose of treating calf diarrhea.