基于网络药理学和试验验证分析小檗碱治疗鸡沙门菌感染的作用机制

doi:10.11843/j.issn.0366-6964.2023.08.039

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (8): 3557-3570.doi: 10.11843/j.issn.0366-6964.2023.08.039

基于网络药理学和试验验证分析小檗碱治疗鸡沙门菌感染的作用机制

张旭梅^1,5, 魏玉荣^2,3, 许丞惠^1,5, 杨彤^1,5, 史慧君^1,5, 付强^1,5, 杨莉^1,4,5*

1. 新疆农业大学动物医学学院, 乌鲁木齐 830052;
2. 新疆动物疫病研究重点实验室, 乌鲁木齐 830013;
3. 新疆畜牧科学院兽医研究所, 乌鲁木齐 830013;
4. 新疆农业大学畜牧学博士后流动站, 乌鲁木齐 830052;
5. 新疆草食动物新药研究与创制重点实验室, 乌鲁木齐 830052

收稿日期:2022-10-31 出版日期:2023-08-23 发布日期:2023-08-22
通讯作者: 杨莉,主要从事兽医药理学研究,E-mail:563289492@qq.com
作者简介:张旭梅(1999-),女,四川乐山人,硕士生,主要从事兽医药理学研究,E-mail:1564546821@qq.com;魏玉荣(1987-),女,甘肃民勤人,研究员,博士,主要从事动物传染病研究,E-mail:weiyu1113@sina.com。
基金资助:
新疆维吾尔自治区自然基金（2022D01B21）；高校科研计划自然科学青年项目（XJEDU2021Y022）；国家自然科学基金（32260902）

To Analyze the Mechanism of Berberine in the Treatment of Salmonella Gallinarum Infection Based on Network Pharmacology and Experimental Verification

ZHANG Xumei^1,5, WEI Yurong^2,3, XU Chenghui^1,5, YANG Tong^1,5, SHI Huijun^1,5, FU Qiang^1,5, YANG Li^1,4,5*

1. College of Animal Medicine, Xinjiang Agricultural University, Urumqi 830052, China;
2. Xinjiang Key Laboratory of Animal Disease Research, Urumqi 830013, China;
3. Institute of Veterinary Medicine, Xinjiang Academy of Animal Science, Urumqi 830013, China;
4. Postdoctoral Station of Animal Husbandry, Xinjiang Agricultural University, Urumqi 830052, China;
5. Xinjiang Key Laboratory of Herbivore New Drug Research and Creation, Urumqi 830052, China

Received:2022-10-31 Online:2023-08-23 Published:2023-08-22

摘要/Abstract

摘要： 采用网络药理学、分子对接、分子动力学模拟等方法，结合沙门菌攻毒试验初步验证小檗碱（berberine，BBR）干预肉鸡肠炎的潜在靶点及作用机制。首先利用PharMapper数据库和Swisstargets数据库预测药物靶点；通过GeneCards数据库收集疾病靶点，并筛选出药物和疾病靶点的交集，将得到的交集靶点构建蛋白质互作网络及GO与KEGG富集分析；对筛选出的核心靶点进行分子对接验证和动力学模拟；后续建立肉鸡肠道炎症模型，观察其肠道组织形态变化，并利用RT-qPCR验证BBR对靶点蛋白mRNA表达量的影响。结果显示：最终筛选得到BBR潜在靶点374个，与肠炎交集174个；GO与KEGG富集分析分别得到197条目和73条目，主要涉及信号转导、蛋白磷酸化过程及PI3K-Akt、趋化因子及Ras等信号通路；分子对接和动力学模拟结果显示，BBR与核心靶点之间均能较好地结合，主要通过疏水作用力、氢键结合；组织切片结果发现，BBR可显著缓解肠道组织损伤；RT-qPCR结果显示，给药后仔鸡盲肠组织中HSP90AA1表达量显著下降（P<0.001），PIK3CA表达量显著上升（P<0.01）。通过网络药理学和试验验证发现，BBR可能通过调节HSP90AA1和PIK3CA表达量来调控肠道炎症的发生发展，可能涉及PI3K-Akt、cAMP、AMPK等信号通路，为深入进行BBR治疗肠炎的作用机制研究提供新思路与新方法。

关键词: 网络药理学, 小檗碱, 肠炎, 分子对接, 作用机制

Abstract: The objective of this study was to preliminaries verify the potential target and mechanism of berberine (BBR) in the intervention of enteritis in broilers by using network pharmacology, molecular docking and molecular dynamics simulation combined with Salmonella challenge test. Firstly, PharMapper database and Swisstargets database were used to predict drug targets. The disease targets were collected through the GeneCards database, and the intersection of drugs and disease targets was screened. The obtained intersection targets were used to construct protein interaction network and GO and KEGG enrichment analysis. Molecular docking verification and kinetic simulation were carried out for the selected core targets. Subsequently, the intestinal inflammation model of broilers was established to observe the morphology changes of intestinal tissue, and the effect of BBR on the mRNA expression of target protein was verified by qPCR test. Finally, 374 potential BBR targets were screened, and 174 overlapped with enteritis. GO and KEGG enrichment analysis obtained 197 items and 73 items, respectively, which mainly involved signal transduction, protein phosphorylation process, PI3K-Akt, chemokine and Ras signaling pathway. The results of molecular docking and kinetic simulation showed that BBR could bind well to the core target, mainly through hydrophobic force and hydrogen bond. The results of tissue section showed that BBR could significantly alleviate intestinal tissue injury. The results of RT-qPCR showed that the expression of HSP90AA1 was significantly decreased (P<0.001), and the expression of PIK3CA was significantly increased (P<0.01) in cecal tissue of the inflammatory model of broilers. Through network pharmacology and experimental verification, it was found that BBR may regulate the occurrence and development of intestinal inflammation by regulating the expression of HSP90AA1 and PIK3CA, which may involve PI3K-Akt, cAMP, AMPK and other signaling pathways, providing new ideas and methods for further research on the mechanism.

Key words: network pharmacology, berberine, enteritis, molecular docking, mechanism of action

中图分类号:

S853.74

张旭梅, 魏玉荣, 许丞惠, 杨彤, 史慧君, 付强, 杨莉. 基于网络药理学和试验验证分析小檗碱治疗鸡沙门菌感染的作用机制[J]. 畜牧兽医学报, 2023, 54(8): 3557-3570.

ZHANG Xumei, WEI Yurong, XU Chenghui, YANG Tong, SHI Huijun, FU Qiang, YANG Li. To Analyze the Mechanism of Berberine in the Treatment of Salmonella Gallinarum Infection Based on Network Pharmacology and Experimental Verification[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3557-3570.

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基于网络药理学和试验验证分析小檗碱治疗鸡沙门菌感染的作用机制

To Analyze the Mechanism of Berberine in the Treatment of Salmonella Gallinarum Infection Based on Network Pharmacology and Experimental Verification

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