基于网络药理学探究党参-甘草提取物增强断奶仔猪免疫功能的作用机制

doi:10.16431/j.cnki.1671-7236.2023.10.042

摘要/Abstract

摘要： 【目的】基于网络药理学结合分子对接技术探究党参-甘草提取物对断奶仔猪免疫功能的影响及其作用机制。【方法】28日龄断奶仔猪随机分为对照组和党参-甘草低、中、高剂量组,每组12只,对照组饲喂基础饲粮,党参-甘草低、中、高剂量组分别在基础饲粮基础上添加0.5%、1%、1.5%党参-甘草提取物,连续饲喂28 d,统计各组断奶仔猪平均日增重、料重比,检测血清中IgA、IgM和IgG含量。通过TCMSP及SwissTargetPrediction数据库筛选党参-甘草潜在活性成分及其作用靶点,通过OMIM和GeneCards数据库检索免疫抑制相关靶点,获得党参-甘草增强免疫作用的潜在作用靶点,利用STRING数据库构建蛋白互作(PPI)网络,通过Metascape数据库进行GO功能和KEGG通路富集分析。利用AutoDock 1.5.6和PyMOL软件对部分关键作用靶点进行分子对接验证和可视化处理,并对其结果进行验证。【结果】与对照组相比,党参-甘草提取物中剂量组断奶仔猪平均日增重、血清中IgA、IgM和IgG含量显著增加,料重比显著降低(P＜0.05)。网络药理学研究表明,党参-甘草主要活性成分可能为槲皮素、山柰酚、木犀草素和蒲公英赛醇,其与免疫抑制疾病交集的关键靶点有95个。党参-甘草调节免疫PPI网络中关联度最高的为丝氨酸/苏氨酸蛋白激酶1(AKT1)、血管内皮生长因子A(VEGFA)、白细胞介素6(IL6)和肿瘤坏死因子α(TNF-α)。GO功能富集分析显示,关键靶点共富集到1 778个条目,主要涉及钙化醇1-单氧酶活性的正向调节、缺氧反应中RNA聚合酶Ⅱ启动子的转录进行正向调节、丝氨酸蛋白酶抑制剂复合物、Bcl-2家族蛋白复合物、肽酶抑制剂复合物等。KEGG通路富集分析显示,党参-甘草调节免疫主要富集在PI3K-Akt、TNF、MAPK等信号通路。分子对接结果显示,槲皮素、山柰酚、木犀草素和蒲公英赛醇与关键作用靶点IL6、VEGFA和TNF-α具有较好结合活性。【结论】饲粮添加党参-甘草提取物可提高断奶仔猪生长性能,增强免疫力,其作用机制可能与干预PI3K-Akt、MAPK、NF-κB等信号通路相关。

关键词: 党参-甘草; 断奶仔猪; 网络药理学; 免疫调节

Abstract: 【Objective】 Based on network pharmacology combined with molecular docking technology,the effects of Codonopsis pilosula-Glycyrrhiza extract on immune function of weaned piglets and its mechanism were investigated.【Method】 Weaned piglets at 28 days of age were randomly divided into control group and Codonopsis pilosula-Glycyrrhiza low,medium and high dose groups, and 12 piglets each group.Piglets in the control group were fed a basal diet,while piglets in the low,medium and high dose groups were fed the basal diet supplemented with 0.5%,1% and 1.5% Codonopsis pilosula-Glycyrrhiza extract for 28 consecutive days,respectively.The average daily gain and feed to gain ratio of weaned piglets in each group were analyzed.The contents of IgA,IgM and IgG in serum were detected.The potential active components and target were screened through TCMSP and SwissTargetPrediction database,and immunosuppressive related targets were searched through OMIM and GeneCards database to obtain the potential target for enhancing the immune effect of Codonopsis pilosula-Glycyrrhiza.The protein-protein interaction (PPI) network was constructed with STRING database,and the GO function and KEGG pathway enrichment analysis were performed with Metascape database.AutoDock 1.5.6 and PyMOL software were used for molecular docking verification and visualization of some key targets,and the results were verified.【Result】 Compared with the control group,the average daily gain, IgA,IgM and IgG contents of serum of weaned piglets in Codonopsis pilosula-Glycyrrhiza extract medium dose group were significantly increased,and the feed to gain ratio was significantly decreased (P ＜ 0.05).Network pharmacological analysis showed that the main active components of Codonopsis pilosula-Glycyrrhiza might be quercetin,kaempferol,luteolin and taraxerol,and there were 95 key targets for their intersection with immunosuppressive diseases.The highest correlation in the immune PPI network was serine/threonine protein kinase 1(AKT1),vascular endothelial growth factor A (VEGFA),interleukin 6 (IL6) and tumor necrosis factor α (TNF-α).GO functional enrichment analysis showed that the key targets were enriched to 1 778 items,mainly related to the positive regulation of calcitriol 1-monooxygenase activity,the transcription of RNA polymerase Ⅱ promoter is positively regulated during hypoxia,serine protease inhibitor complex,Bcl-2 family protein complex,peptidase inhibitor complex,etc.KEGG pathway enrichment analysis showed that the immune regulation of Codonopsis pilosula-Glycyrrhiza was mainly enriched in PI3K-Akt,TNF,MAPK signaling pathways.Molecular docking results showed that quercetin,kaempferol,luteolin and taraxerol had good binding activity with key targets IL6,VEGFA and TNF-α.【Conclusion】 Codonopsis pilosula-Glycyrrhiza extract could improve the growth performance and immunity of weaned piglets,which was closely related to the intervention of PI3K-Akt,MAPK and NF-κB signaling pathways.

Key words: Codonopsis pilosula-Glycyrrhiza; weaned piglets; network pharmacological; immunomodulation

中图分类号:

S853.74

蔡燕子, 袁子文, 张兴平, 张丰裕, 祝亚梅, 史艳, 姜立冬, 华永丽, 魏彦明. 基于网络药理学探究党参-甘草提取物增强断奶仔猪免疫功能的作用机制[J]. 中国畜牧兽医, 2023, 50(10): 4279-4291.

CAI Yanzi, YUAN Ziwen, ZHANG Xingping, ZHANG Fengyu, ZHU Yamei, SHI Yan, JIANG Lidong, HUA Yongli, WEI Yanming. Study on Mechanism of Codonopsis pilosula-Glycyrrhiza Extract in Enhancing Immune Functions of Weaned Piglets Based on Pharmacology[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 4279-4291.

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