青黛复方制剂调节猪免疫力的作用机制分析

Analysis on mechanism of Qingdai compound preparation regulating immunity in pigs

试验旨在应用网络药理学方法,探究青黛复方制剂调节猪免疫力的分子机制。从Tcmsp数据库中筛查青黛复方制剂含有的化合物,得到有效活性成分,预测有效活性成分作用的靶基因,制作“活性成分-靶基因”网络分析,筛出主要活性成分。通过Omim等数据库分析免疫相关基因,与活性成分的靶基因取交集,得到复方制剂靶向的猪免疫相关基因。利用String数据库构建蛋白互作网络图(PPI),运用Cytoscape筛选PPI的核心蛋白;通过David数据库分析靶基因富集的基因功能和KEGG通路;用Autodock和PyMol进行分子对接,验证核心活性成分与核心靶点蛋白的相互作用。结果显示:青黛复方制剂含有330种化合物,其中有效活性成分为62种,主要的活性物质有β-谷甾醇、豆甾醇、常春藤皂苷元、山柰酚、异牡荆素等10种;有效活性成分作用于64个猪免疫相关基因以及SLC6A4、ESR1、TNF等20种核心靶蛋白,通过IL-17信号通路、TNF信号传导途径、NOD样受体信号传导途径等调节天然免疫和特异性免疫系统功能,提高香猪的抵抗力。青黛复方制剂主要通过β-谷甾醇、异牡荆素等主要活性成分,影响白细胞介素-2(IL-2)、肿瘤坏死因子-α(IFN-α)、干扰素-γ(IFN-γ)的表达水平。分子对接分析表明,这些活性成分能够与靶蛋白有效结合,可为中药增强机体免疫力的物质基础和作用机制等研究提供参考。同时,网络药理学分析结果可为全面解析青黛复方制剂增强猪免疫力的作用机制提供依据。

The aim of the experiment was to explore the molecular mechanism of Qingdai compound preparation regulating immunity in pigs by using network pharmacology.The compounds contained in Qingdai compound preparation were screened from the Tcmsp database to obtain the active active ingredients,and the target genes for the action of the active active ingredients were predicted.The'active ingredient-target gene'network analysis was made to screen out the main active ingredients.The immune-related genes were analyzed through Omim database and intersected with the target genes of active ingredients to obtain the porcine immune-related genes targeted by the compound preparation.The protein interaction network map(PPI)was constructed using String database and Cytoscape was used to screen the core proteins of PPI.The gene function and KEGG pathway of target gene enrichment were analyzed by David database.Molecular docking with Autodock and PyMol was performed to verify the interaction between core active ingredients and core target proteins.The results showed that Qingdai compound preparation contained 330 compounds,of which 62 were active ingredients,and 10 were main active substances,includingβ-sitosterol,stigmasterol,iviogenin,kaempferol,isovitexin,etc.The active ingredient acts on 64 porcine immune-related genes and 20 core target proteins such as SLC6A4,ESR1,TNF,etc.It regulates the natural immunity and specific immune system functions through IL-17 signaling pathway,TNF signaling pathway,NOD-like receptor signaling pathway,etc.,and improves the resistance of porcine pigs.Qingdai compound preparation can affect the expression of IL-2,IFN-α,IFN-γand other inflammatory factors throughβ-sitosterol,isovitexin and other main active ingredients.Macromolecular docking analysis showed that these active ingredients could effectively bind to target proteins,which could provide a reference for the research on the material basis and mechanism of traditional Chinese medicine to enhance immunity.At the same time,the results of network pharmacology analysis can provide a theoretical basis for a comprehensive analysis of the mechanism of Indigo Naturalis Polypill in enhancing pig immunity.