Volume 41 Issue 4
Jul.  2020
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Article Contents
Lei Liu, Da-Wei Song, Guang-Lu Liu, Li-Peng Shan, Tian-Xiu Qiu, Jiong Chen. Hydroxycoumarin efficiently inhibits spring viraemia of carp virus infection in vitro and in vivo. Zoological Research, 2020, 41(4): 395-409. doi: 10.24272/j.issn.2095-8137.2020.037
Citation: Lei Liu, Da-Wei Song, Guang-Lu Liu, Li-Peng Shan, Tian-Xiu Qiu, Jiong Chen. Hydroxycoumarin efficiently inhibits spring viraemia of carp virus infection in vitro and in vivo. Zoological Research, 2020, 41(4): 395-409. doi: 10.24272/j.issn.2095-8137.2020.037

Hydroxycoumarin efficiently inhibits spring viraemia of carp virus infection in vitro and in vivo

doi: 10.24272/j.issn.2095-8137.2020.037
Funds:  This work was supported by the Program for the National Natural Science Foundation of China (31902410), Natural Science Foundation of Ningbo City of China (2019A610419), General Scientific Research Project of Education of Zhejiang Province (Y201839187), Scientific Innovation Team Project of Ningbo (2015C110018), Scientific Research Fund of Ningbo University (XYL19017), and K.C. Wong Magna Fund in Ningbo University
More Information
  • Corresponding author: E-mail: jchen1975@163.com
  • Received Date: 2020-02-10
  • Accepted Date: 2020-04-24
  • Available Online: 2020-05-10
  • Publish Date: 2020-07-18
  • Spring viremia of carp virus (SVCV) causes devastating losses in aquaculture. Coumarin has an advantageous structure for the design of novel antiviral agents with high affinity and specificity. In this study, we evaluated a hydroxycoumarin medicine, i.e., 7-(6-benzimidazole) coumarin (C10), regarding its anti-SVCV effects in vitro and in vivo. Results showed that up to 12.5 mg/L C10 significantly inhibited SVCV replication in the epithelioma papulosum cyprini (EPC) cell line, with a maximum inhibitory rate of >97%. Furthermore, C10 significantly reduced cell death and relieved cellular morphological damage in SVCV-infected cells. Decreased mitochondrial membrane potential (ΔΨm) also suggested that C10 not only protected mitochondria, but also reduced apoptosis in SVCV-infected cells. For in vivo studies, intraperitoneal injection of C10 resulted in an anti-SVCV effect and substantially enhanced the survival rate of virus-infected zebrafish. Furthermore, C10 significantly enhanced antioxidant enzyme activities and decreased reactive oxygen species (ROS) to maintain antioxidant-oxidant balance within the host, thereby contributing to inhibition of SVCV replication. The up-regulation of six interferon (IFN)-related genes also demonstrated that C10 indirectly activated IFNs for the clearance of SVCV in zebrafish. This was beneficial for the continuous maintenance of antiviral effects because of the low viral loads in fish. Thus, C10 is suggested as a therapeutic agent with great potential against SVCV infection in aquaculture.

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  • [1]
    Adamek M, Rakus KŁ, Chyb J, Brogden G, Huebner A, Irnazarow I, Steinhagen D. 2012. Interferon type I responses to virus infections in carp cells: in vitro studies on Cyprinid herpesvirus 3 and Rhabdovirus carpio infections. Fish & Shellfish Immunology, 33(3): 482−493.
    [2]
    Aggad D, Mazel M, Boudinot P, Mogensen KE, Hamming OJ, Hartmann R, Kotenko S, Herbomel P, Lutfalla G, Levraud JP. 2009. The two groups of zebrafish virus-induced interferons signal via distinct receptors with specific and shared chains. The Journal of Immunology, 183(6): 3924−3931. doi:  10.4049/jimmunol.0901495
    [3]
    Ahne W, Bjorklund HV, Essbauer S, Fijan N, Kurath G, Winton JR. 2002. Spring viremia of carp (SVC). Diseases of Aquatic Organisms, 52(3): 261−272.
    [4]
    Ashraf U, Lu YA, Lin L, Yuan JF, Wang M, Liu XQ. 2016. Spring viraemia of carp virus: recent advances. Journal of General Virology, 97(5): 1037−1051. doi:  10.1099/jgv.0.000436
    [5]
    Balmer BF, Powers RL, Zhang TH, Lee J, Vigant F, Lee B, Jung ME, Purcell MK, Snekvik K, Aguilar HC. 2017. Inhibition of an aquatic rhabdovirus demonstrates promise of a broad-spectrum antiviral for use in aquaculture. Journal of Virology, 91(4): e02181-16. doi:  10.1128/JVI.02181-16
    [6]
    Balmer BF, Getchell RG, Powers RL, Lee J, Zhang TH, Jung ME, Purcell MK, Snekvik K, Aguilar HC. 2018. Broad-spectrum antiviral JL122 blocks infection and inhibits transmission of aquatic rhabdoviruses. Virology, 525: 143−149. doi:  10.1016/j.virol.2018.09.009
    [7]
    Bourinbaiar AS, Tan X, Nagorny R. 1993. Effect of the oral anticoagulant, warfarin, on HIV-1 replication and spread. AIDS, 7(1): 129−130. doi:  10.1097/00002030-199301000-00022
    [8]
    Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2): 248−254. doi:  10.1016/0003-2697(76)90527-3
    [9]
    Chen HY, Liu WT, Wu SY, Chiou PP, Li YH, Chen YC, Lin GH, Lu MW, Wu JL. 2015. RIG-I specifically mediates group II type I IFN activation in nervous necrosis virus infected zebrafish cells. Fish & Shellfish Immunology, 43(2): 427−435.
    [10]
    Chen ZY, Liu H, Li ZQ, Wang M, Zhang QY. 2006. Detection of viral pathogen from diseased common carp (Cyprinus carpio) by infectious tests. Journal of Fishery Sciences of China, 13(4): 617−623. (in Chinese)
    [11]
    Cobbina SJ, Xu H, Zhao T, Mao GH, Zhou ZX, Wu XS, Liu HY, Zou YM, Wu XY, Yang LQ. 2015. A multivariate assessment of innate immune-related gene expressions due to exposure to low concentration individual and mixtures of four kinds of heavy metals on zebrafish (Danio rerio) embryos. Fish & Shellfish Immunology, 47(2): 1032−1042.
    [12]
    Dikkeboom AL, Radi C, Toohey-Kurth K, Marcquenski S, Engel M, Goodwin AE, Way K, Stone DM, Longshaw C. 2004. First report of spring viremia of carp virus (SVCV) in wild common carp in North America. Journal of Aquatic Animal Health, 16(4): 169−178. doi:  10.1577/H03-064.1
    [13]
    Encinas P, Garcia-Valtanen P, Chinchilla B, Gomez-Casado E, Estepa A, Coll J. 2013. Identification of multipath genes differentially expressed in pathway-targeted microarrays in zebrafish infected and surviving spring viremia carp virus (SVCV) suggest preventive drug candidates. PloS One, 8: e73553. doi:  10.1371/journal.pone.0073553
    [14]
    Fijan N. 1984. Vaccination of fish in European pond culture: prospects and constraints. Symposia Biologica Hungaria, 23: 233−241.
    [15]
    Forlenza M, de Carvalho Dias JD, Veselý T, Pokorová D, Savelkoul HFJ, Wiegertjes GF. 2008. Transcription of signal-3 cytokines, IL-12 and IFNαβ, coincides with the timing of CD8αβ up-regulation during viral infection of common carp (Cyprinus carpio L.). Molecular Immunology, 45(6): 1531−1547. doi:  10.1016/j.molimm.2007.10.010
    [16]
    Fujioka H, Yamasaki K, Furusawa K, Tamura K, Oguro K, Kurihara S, Seki S, Oshima SI, Imajoh M. 2015. Prevalence and characteristics of Cyprinid herpesvirus 3 (CyHV-3) infection in common carp (Cyprinus carpio L.) inhabiting three rivers in Kochi Prefecture, Japan. Veterinary Microbiology, 175(2-4): 362−368. doi:  10.1016/j.vetmic.2014.12.002
    [17]
    Garaci E, Palamara AT, Ciriolo MR, D'Agostini C, Ab del-Latif MS, Aquaro S, Lafavia E, Rotilio G. 1997. Intracellular GSH content and HIV replication in human macrophages. Journal of Leukocyte Biology, 62(1): 54−59. doi:  10.1002/jlb.62.1.54
    [18]
    Garver KA, Dwilow AG, Richard J, Booth TF, Beniac DR, Souter BW. 2007. First detection and confirmation of spring viraemia of carp virus in common carp, Cyprinus carpio L., from Hamilton Harbour, Lake Ontario, Canada. Journal of Fish Diseases, 30(11): 665−671. doi:  10.1111/j.1365-2761.2007.00851.x
    [19]
    Hassan MZ, Osman H, Ali MA, Ahsan MJ. 2016. Therapeutic potential of coumarins as antiviral agents. European Journal of Medicinal Chemistry, 123: 236−255. doi:  10.1016/j.ejmech.2016.07.056
    [20]
    Huang L, Kashiwada Y, Cosentino LM, Fan S, Chen CH, McPhail AT, Fujioka T, Mihashi K, Lee KH. 1994a. Anti-AIDS agents. 15. Synthesis and anti-HIV activity of dihydroseselins and related analogs. Journal of Medicinal Chemistry, 37(23): 3947−3955. doi:  10.1021/jm00049a014
    [21]
    Huang L, Kashiwada Y, Cosentino LM, Fan S, Lee KH. 1994b. 3’,4’-Di-o-(-)-camphanoyl-(+)-ciskhellactone and related compounds: A. New class of potent anti-HIV agents. Bioorganic & Medicinal Chemistry Letters, 4(4): 593−598.
    [22]
    Koutná M, Veselý T, Psikal I, Hůlová J. 2003. Identification of spring viraemia of carp virus (SVCV) by combined RT-PCR and nested PCR. Diseases of Aquatic Organisms, 55(3): 229−235.
    [23]
    Liu GL, Wang CJ, Wang HF, Zhu LL, Zhang H, Wang YS, Pei CY, Liu L. 2019a. Antiviral efficiency of a coumarin derivative on spring viremia of carp virus in vivo. Virus Research, 268: 11−17. doi:  10.1016/j.virusres.2019.05.007
    [24]
    Liu L, Hu Y, Lu JF, Wang GX. 2019b. An imidazole coumarin derivative enhances the antiviral response to spring viremia of carp virus infection in zebrafish. Virus Research, 263: 112−118. doi:  10.1016/j.virusres.2019.01.009
    [25]
    Liu L, Hu Y, Shen YF, Wang GX, Zhu B. 2017. Evaluation on antiviral activity of coumarin derivatives against spring viraemia of carp virus in epithelioma papulosum cyprini cells. Antiviral Research, 144: 173−185. doi:  10.1016/j.antiviral.2017.06.007
    [26]
    Liu L, Qiu TX, Song DW, Shan LP, Chen J. 2020. Inhibition of a novel coumarin on an aquatic rhabdovirus by targeting the early stage of viral infection demonstrates potential application in aquaculture. Antiviral Research, 174: 104672. doi:  10.1016/j.antiviral.2019.104672
    [27]
    Liu L, Shen YF, Hu Y, Lu JF. 2018. Antiviral effect of 7-(4-benzimidazole-butoxy)-coumarin on rhabdoviral clearance via Nrf2 activation regulated by PKCα/β phosphorylation. Fish & Shellfish Immunology, 83: 386−396.
    [28]
    Liu L, Zhu B, Wang GX. 2015. Azoxystrobin-induced excessive reactive oxygen species (ROS) production and inhibition of photosynthesis in the unicellular green algae Chlorella vulgaris. Environmental Science and Pollution Research, 22(10): 7766−7775. doi:  10.1007/s11356-015-4121-7
    [29]
    Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25(4): 402−408. doi:  10.1006/meth.2001.1262
    [30]
    López-Muñoz A, Roca FJ, Meseguer J, Mulero V. 2009. New insights into the evolution of IFNs: zebrafish group II IFNs induce a rapid and transient expression of IFN-dependent genes and display powerful antiviral activities. The Journal of Immunology, 182(6): 3440−3449. doi:  10.4049/jimmunol.0802528
    [31]
    Michalek RD, Pellom ST, Holbrook BC, Grayson JM. 2008. The requirement of reactive oxygen intermediates for lymphocytic choriomeningitis virus binding and growth. Virology, 379(2): 205−212. doi:  10.1016/j.virol.2008.07.004
    [32]
    Miller O, Fuller FJ, Gebreyes WA, Lewbart GA, Shchelkunov IS, Shivappa RB, Joiner C, Woolford G, Stone DM, Dixon PF, Raley ME, Levine JF. 2007. Phylogenetic analysis of spring virema of carp virus reveals distinct subgroups with common origins for recent isolates in North America and the UK. Diseases of Aquatic Organisms, 76(3): 193−204.
    [33]
    Neyts J, De Clercq E, Singha R, Chang YH, Das AR, Chakraborty SK, Hong SC, Tsay SC, Hsu MH, Hwu JR. 2009. Structure-activity relationship of new anti-hepatitis C virus agents: heterobicycle-coumarin conjugates. Journal of Medicinal Chemistry, 52(5): 1486−1490. doi:  10.1021/jm801240d
    [34]
    Peng XM, Damu GLV, Zhou CH. 2013. Current developments of coumarin compounds in medicinal chemistry. Current Pharmaceutical Design, 19(21): 3884−3930. doi:  10.2174/1381612811319210013
    [35]
    Penta S. 2015. Advances in Structure and Activity Relationship of Coumarin Derivatives. Amsterdam: Academic Press.
    [36]
    Petty BD, Riggs AC, Klinger R, Yanong RP, Francis-Floyd R. 2002. Spring viremia of carp. Fact Sheet VM-142. Gainesville, FL: University of Florida Cooperative Extension Service, University of Florida.
    [37]
    Pizzi M. 1950. Sampling variation of the fifty percent end-point, determined by the Reed-Muench (Behrens) method. Human Biology, 22(3): 151−190.
    [38]
    Qiu TX, Song DW, Shan LP, Liu GL, Liu L. 2020. Potential prospect of a therapeutic agent against spring viraemia of carp virus in aquaculture. Aquaculture, 515: 734558. doi:  10.1016/j.aquaculture.2019.734558
    [39]
    Robertsen B. 2006. The interferon system of teleost fish. Fish & Shellfish Immunology, 20(2): 172−191.
    [40]
    Sancho R, Márquez N, Gómez-Gonzalo M, Calzado MA, Bettoni G, Coiras MT, Alcamí J, López-Cabrera M, Appendino G, Muñoz E. 2004. Imperatorin inhibits HIV-1 replication through an Sp1-dependent pathway. The Journal of Biological Chemistry, 279(36): 37349−37359. doi:  10.1074/jbc.M401993200
    [41]
    Shao JH, Huang J, Guo YN, Li LJ, Liu X1, Chen XX, Yuan JF. 2016. Up-regulation of nuclear factor E2-related factor 2 (Nrf2) represses the replication of SVCV. Fish & Shellfish Immunology, 58: 474−482.
    [42]
    Shen YF, Hu Y, Zhang Z, Liu L, Chen C, Tu X, Wang GX, Zhu B. 2019. Saikosaponin D efficiently inhibits SVCV infection in vitro and in vivo. Aquaculture, 504: 281−290. doi:  10.1016/j.aquaculture.2019.02.014
    [43]
    Shen YF, Liu L, Feng CZ, Hu Y, Chen C, Wang GX, Zhu B. 2018. Synthesis and antiviral activity of a new coumarin derivative against spring viraemia of carp virus. Fish & Shellfish Immunology, 81: 57−66.
    [44]
    Song DW, Liu L, Shan LP, Qiu TX, Chen J, Chen JP. 2020. Therapeutic potential of phenylpropanoid-based small molecules as anti-SVCV agents in aquaculture. Aquaculture, 526: 735349. doi:  10.1016/j.aquaculture.2020.735349
    [45]
    Stubbs MT, Bode W. 1993. A player of many parts: the spotlight falls on thrombin’s structure. Thrombosis Research, 69(1): 1−58. doi:  10.1016/0049-3848(93)90002-6
    [46]
    Teng Y, Liu H, Lv JQ, Fan WH, Zhang QY, Qin QW. 2007. Characterization of complete genome sequence of the spring viremia of carp virus isolated from common carp (Cyprinus carpio) in China. Archives of Virology, 152(8): 1457−1465. doi:  10.1007/s00705-007-0971-8
    [47]
    Wang YD, Zhang H, Lu YA, Wang F, Liu LY, Liu JX, Liu XQ. 2017. Comparative transcriptome analysis of zebrafish (Danio rerio) brain and spleen infected with spring viremia of carp virus (SVCV). Fish & Shellfish Immunology, 69: 35−45.
    [48]
    Wang ZG, Tang XB, Li YM, Leu C, Guo L, Zheng XD, Zhu DL. 2008. 20-Hydroxyeicosatetraenoic acid inhibits the apoptotic responses in pulmonary artery smooth muscle cells. European Journal of Pharmacology, 588(1): 9−17. doi:  10.1016/j.ejphar.2008.03.045
    [49]
    Wei XX, Li XZ, Zheng XC, Jia P, Wang JJ, Yang XL, Yu L, Shi XJ, Tong GX, Liu H. 2016. Toll-like receptors and interferon associated immune factors responses to spring viraemia of carp virus infection in common carp (Cyprinus carpio). Fish & Shellfish Immunology, 55: 568−576.
    [50]
    Workenhe ST, Rise ML, Kibenge MJT, Kibenge FSB. 2010. The fight between the teleost fish immune response and aquatic viruses. Molecular Immunology, 47: 2525−2536. doi:  10.1016/j.molimm.2010.06.009
    [51]
    Yuan JF, Su N, Wang M, Xie P, Shi ZL, Li LJ. 2012. Down-regulation of heme oxygenase-1 by SVCV infection. Fish & Shellfish Immunology, 32(2): 301−306.
    [52]
    Yuan JF, Yang Y, Nie HH, Li LJ, Gu WG, Lin L, Zou M, Liu XQ, Wang M, Gu ZM. 2014. Transcriptome analysis of epithelioma papulosum cyprini cells after SVCV infection. BMC Genomics, 15(1): 935. doi:  10.1186/1471-2164-15-935
    [53]
    Zhang YN, Li ZC, Ge XN, Guo X, Yang HC. 2011. Autophagy promotes the replication of encephalomyocarditis virus in host cells. Autophagy, 7(6): 613−628. doi:  10.4161/auto.7.6.15267
    [54]
    Zhao LT, Qi L, Li C, Li LJ, Jin L, Yuan JF. 2018. SVCV impairs mitochondria complex Ⅲ resulting in accumulation of hydrogen peroxide. Fish & Shellfish Immunology, 75: 58−65.
    [55]
    Zorova LD, Popkov VA, Plotnikov EY, Silachev DN, Pevzner IB, Jankauskas SS, Babenko VA, Zorov SD, Balakireva AV, Juhaszova M, Sollott SJ, Zorov DB. 2018. Mitochondrial membrane potential. Analytical Biochemistry, 552: 50−59. doi:  10.1016/j.ab.2017.07.009
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