基于网络药理学和分子对接分析苦参碱抗PRRSV的作用机制

doi:10.11843/j.issn.0366-6964.2021.03.025

摘要/Abstract

摘要： 本研究旨在利用网络药理学及分子对接技术从抗炎角度探讨苦参碱抗猪繁殖与呼吸综合征病毒（PRRSV）的作用机制。利用PharmMapper服务器和Genecards疾病数据库获得苦参碱发挥抗炎作用的潜在靶点；使用STRING在线分析平台结合Cytoscape软件构建靶蛋白相互作用（protein-protein interaction，PPI）网络并进行拓扑学分析筛选关键靶点；利用DAVID数据库对靶点进行基因本体论GO富集分析以及KEGG通路富集分析，并使用Cytoscape软件进行“药物-靶点-通路”网络图的构建；使用Autodock Vina软件进行分子对接，选择最佳的结合靶点。网络分析结果表明，苦参碱抗炎的潜在靶点有23个；蛋白互作网络提示，IGFⅠ、MAPK8、CASP3、NR3C1可能是苦参碱抗炎的核心靶点；GO富集分析得到116个细胞生物学过程，KEGG通路富集分析得到47条相关信号通路；分子对接显示，MAPK8与苦参碱有较好亲和力，可能是苦参碱发挥抗炎作用的主要靶点。苦参碱可能通过作用于MAPK8这一关键蛋白发挥抗炎作用，进而达到抗PRRSV的作用。

关键词: 苦参碱, 网络药理学, 抗炎, 靶点

Abstract: Based on network pharmacology and molecular docking technology, this study explored the anti-PRRSV mechanism of matrine from the perspective of anti-inflammatory. PharmMapper server and Genecards disease database were used to obtain the potential targets of matrine; STRING online analysis platform combined with Cytoscape software was used to construct the target protein-protein interaction (PPI) network and perform topological analysis to screen the key targets; DAVID database was used to perform gene ontology GO enrichment analysis and KEGG pathway enrichment analysis of the targets, and Cytoscape software was used to construct the "drug-target-pathway" network diagram; Autodock Vina software was used for molecular docking to select the best binding targets. The results of network analysis showed that there were 23 potential targets of matrine; the protein interaction network suggested that IGFⅠ, MAPK8, CASP3, and NR3C1 may be the core targets of matrine; 116 cell biological processes were obtained by GO enrichment analysis, and 47 related signaling pathways were obtained by KEGG pathway enrichment analysis; molecular docking showed that MAPK8 had a good affinity to matrine and maybe the main target of matrine exerting its effect. Matrine may play an anti-inflammatory role by acting on the key protein of MAPK8, thus achieving anti-PRRSV effect.

Key words: matrine, network pharmacology, anti-inflammatory, target

中图分类号:

S853.74

郝江花, 孙娜, 孙盼盼, 孙耀贵, 范阔海, 尹伟, 李宏全. 基于网络药理学和分子对接分析苦参碱抗PRRSV的作用机制[J]. 畜牧兽医学报, 2021, 52(3): 809-819.

HAO Jianghua, SUN Na, SUN Panpan, SUN Yaogui, FAN Kuohai, YIN Wei, LI Hongquan. A Study on Mechanism of Matrine against PRRSV Based on Network Pharmacology and Molecular Docking[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 809-819.

参考文献 30

[1]	刘旭, 梁跃辉, 赵晓秋, 等. 苦参碱类TNF-α抑制剂的合成、抗炎活性评价和分子对接研究[J]. 计算机与应用化学, 2016, 33(5):521-524.LIU X, LIANG Y H, ZHAO X Q, et al. Synthesis of TNF-α inhibitors, anti inflammatory activity evaluation and molecular docking study of matrine derivatives[J]. Computers and Applied Chemistry, 2016, 33(5):521-524. (in Chinese)
[2]	赵昕, 孙娜, 白喜云, 等. 苦参碱对PRRSV的体外抑制作用及其机制[J]. 中国兽医学报, 2013, 33(6):808-812.ZHAO X, SUN N, BAI X Y, et al. Inhibitory action and mechanism of matrine against porcine reproductive and respiratory syndrome virus in vitro[J]. Chinese Journal of Veterinary Science, 2013, 33(6):808-812. (in Chinese)
[3]	SUN N, SUN P P, LV H P, et al. Matrine displayed antiviral activity in porcine alveolar macrophages co-infected by porcine reproductive and respiratory syndrome virus and porcine circovirus type 2[J]. Sci Rep, 2016, 6:24401.
[4]	SUN N, WANG Z W, WU C H, et al. Antiviral activity and underlying molecular mechanisms of matrine against porcine reproductive and respiratory syndrome virus in vitro[J]. Res Vet Sci, 2014, 96(2):323-327.
[5]	ZHANG B, LIU Z Y, LI Y Y, et al. Antiinflammatory effects of matrine in LPS-induced acute lung injury in mice[J]. Eur J Pharm Sci, 2011, 44(5):573-579.
[6]	LIOU C J, LAI Y R, CHEN Y L, et al. Matrine attenuates COX-2 and ICAM-1 expressions in human lung epithelial cells and prevents acute lung injury in LPS-induced mice[J]. Mediators Inflamm, 2016, 2016:3630485.
[7]	NIU Y J, DONG Q M, LI R H. Matrine regulates Th1/Th2 cytokine responses in rheumatoid arthritis by attenuating the NF-κB signaling[J]. Cell Bio Int, 2017, 41(6):611-621.
[8]	CHANG J L, HU S P, WANG W Y, et al. Matrine inhibits prostate cancer via activation of the unfolded protein response/endoplasmic reticulum stress signaling and reversal of epithelial to mesenchymal transition[J]. Mol Med Rep, 2018, 18(1):945-957.
[9]	孙娜. 苦参碱抗PRRSV/PCV2共感染及其作用机制[D]. 晋中:山西农业大学, 2013.SUN N. Antiinfection effect of matrine on PRRSV/PCV2 co-infection and its mechanism[D]. Jinzhong:Shanxi Agricultural University, 2013. (in Chinese)
[10]	LIU K, LI Y M, ZHOU B, et al. A conjugate protein containing HIV TAT, ISG20, and a PRRSV polymerase binding inhibits PRRSV replication and may be a novel therapeutic platform[J]. Res Vet Sci, 2017, 113:13-20.
[11]	孙盼盼. 苦参碱抑制PAMs分泌IL-1β的机理及联合用药研究[D]. 晋中:山西农业大学, 2019.SUN P P. Study on the mechanism of matrine inhibiting PAMs secrete IL-1β and its combined effect with antitiotics[D]. Jinzhong:Shanxi Agricultural University, 2019. (in Chinese)
[12]	孙盼盼, 侯雅鑫, 张宇瞳, 等. 苦参碱对PRRSV/LPS共刺激诱导小鼠炎症反应的作用[J]. 中国兽医杂志, 2019, 55(8):40-45.SUN P P, HOU Y X, ZHANG Y T, et al. Effect of matrine on inflammatory response induced by PRRSV/LPS co-stimulation in mice[J]. Chinese Journal of Veterinary Medicine, 2019, 55(8):40-45. (in Chinese)
[13]	汝锦龙. 中药系统药理学数据库和分析平台的构建和应用[D]. 杨凌:西北农林科技大学, 2015.RU J L. Construction and utilization of traditional Chinese medicine systems pharmacology database and analysis platform[D]. Yangling:Northwest A&F University, 2015. (in Chinese)
[14]	周文霞. 网络药理学的研究进展和发展前景[J]. 中国药理学与毒理学杂志, 2015, 29(5):760-762.ZHOU W X. Research progress and prospect of network pharmacology[J]. Chinese Journal of Pharmacology and Toxicology, 2015, 29(5):760-762. (in Chinese)
[15]	王琳珊, 靳会欣, 董占军. 系统药理学研究方法在中药不良反应研究中的应用进展[J]. 中国药房, 2017, 28(35):5033-5036.WANG L S, JIN H X, DONG Z J. Progress in the application of systematic pharmacology research methods in the study of adverse reactions of traditional Chinese medicine[J]. China Pharmacy, 2017, 28(35):5033-5036. (in Chinese)
[16]	黄国东, 孙秀玉, 陈书, 等. 网络药理学在国内中药复方研究的应用概况[J]. 广西中医药大学学报, 2016, 19(1):104-107.HUANG G D, SUN X Y, CHEN S, et al. Application of network pharmacology in Chinese traditional medicine compound research[J]. Journal of Guangxi University of Chinese Medicine, 2016, 19(1):104-107. (in Chinese)
[17]	沈汶, 蒋卓霖, 吴雨晴, 等. 基于网络药理学探究银黄药对抗病毒作用机制[J]. 中成药, 2020, 42(5):1329-1333.SHEN W, JIANG Z L, WU Y Q, et al. Exploring the mechanism of antiviral effect of Yinhuang medicine based on network pharmacology[J]. Chinese Traditional Patent Medicine, 2020, 42(5):1329-1333. (in Chinese)
[18]	崔琳琳, 宋亚刚, 苗明三. 基于网络药理学和分子对接的陈皮干预COVID-19的可能机制[J]. 中药药理与临床, 2020, 36(5):28-33.CUI L L, SONG Y G, MIAO M S. Possible mechanism of Citri Reticulatae Pericarpium intervening on COVID-19 based on network pharmacology and molecular docking[J]. Pharmacology and Clinics of Chinese Materia Medica, 2020, 36(5):28-33. (in Chinese)
[19]	周丽娟. IGFⅠ在流感病毒介导的炎性肺损伤中的作用机制研究[D]. 淮北:淮北师范大学, 2017.ZHOU L J. Study on action mechanism of IGFⅠ in inflammatory pulmonary injury induced by influenza virus[D]. Huaibei:Huaibei Normal University, 2017. (in Chinese)
[20]	LI G P, ZHOU L J, ZHANG C, et al. Insulin-like growth factor 1 regulates acute inflammatory lung injury mediated by influenza virus infection[J]. Front Microbiol, 2019, 10:2541.
[21]	苗光新, 李蒙蒙, 吴晓光. 山楂叶总黄酮对脑缺血大鼠Caspase-3蛋白及炎症因子表达的影响[J]. 中国老年学杂志, 2019, 39(10):2487-2490.MIAO G X, LI M M, WU X G. Effects of hawthorn leaf flavonoids on expression of Caspase-3 protein and inflammatory factors in cerebral ischemic rats[J]. Chinese Journal of Gerontology, 2019, 39(10):2487-2490. (in Chinese)
[22]	SUZUKI T, ICHⅡ O, NAKAMURA T, et al. Immune-associated renal disease found in caspase 3-deficient mice[J]. Cell Tissue Res, 2020, 379(2):323-335.
[23]	杜慧. NR3C1基因多态性与高原肺水肿遗传易感性研究[D]. 西宁:青海大学, 2018.DU H. Genetic susceptibility study between NR3C1 gene polymorphisms and high altitude pulmonary edema[D]. Xining:Qinghai University, 2018.
[24]	MYLKA V, DECKERS J, RATMAN D, et al. The autophagy receptor SQSTM1/p62 mediates anti-inflammatory actions of the selective NR3C1/glucocorticoid receptor modulator compound A (CpdA) in macrophages[J]. Autophagy, 2018, 14(12):2049-2064.
[25]	SHAH S, KING E M, CHANDRASEKHAR A, et al. Roles for the mitogen-activated protein kinase (MAPK) phosphatase, DUSP1, in feedback control of inflammatory gene expression and repression by dexamethasone[J]. J Biol Chem, 2014, 289(19):13667-13679.
[26]	KUMAR A, SINGH U K, KINI S G, et al. JNK pathway signaling:a novel and smarter therapeutic targets for various biological diseases[J]. Future Med Chem, 2015, 7(15):2065-2086.
[27]	万智双, 熊丁, 曹宸. miR-433-3p靶向MAPK8对肝癌细胞MHCC97H增殖、凋亡和迁移的调控作用[J]. 中国免疫学杂志, 2020, 36(1):57-62.WAN Z S, XIONG D, CAO C. Regulation of miR-433-3p targeting MAPK8 on proliferation, apoptosis and migration of hepatoma cells MHCC97H[J]. Chinese Journal of Immunology, 2020, 36(1):57-62. (in Chinese)
[28]	雷莹莹. 亲环蛋白A对PRRSV增殖的影响及其作用机制研究[D]. 武汉:华中农业大学, 2014.LEI Y Y. The effects and underlying mechanisms of Cyclophilin A on PRRSV replication[D]. Wuhan:Huazhong Agricultural University, 2014. (in Chinese)
[29]	余志彬. 猪繁殖与呼吸综合征病毒(PRRSV)诱导白介素-12(IL-12)产生的分子机制研究[D]. 武汉:华中农业大学, 2016.YU Z B. Study on the mechanism of interleukin-12(IL-12) production induced by porcine reproductive and respiratory syndrome virus (PRRSV)[D]. Wuhan:Huazhong Agricultural University, 2016. (in Chinese)
[30]	LIU Y H, DU Y P, WANG H L, et al. Porcine reproductive and respiratory syndrome virus (PRRSV) up-regulates IL-8 expression through TAK-1/JNK/AP-1 pathways[J]. Virology, 2017, 506:64-72.