In vitro antiviral activity and underlying molecular mechanisms of dipotassium glycyrrhetate against porcine reproductive and respiratory syndrome virus
Free accessResearch articleFirst published online November, 2013
In vitro antiviral activity and underlying molecular mechanisms of dipotassium glycyrrhetate against porcine reproductive and respiratory syndrome virus
Porcine reproductive and respiratory syndrome virus (PRRSV) has caused large economic losses in the swine industry. Currently, there is no effective way to prevent PRRSV infection. In this study, we investigated the inhibitory effect of dipotassium glycyrrhetate (DG), a derivative of glycyrrhetinic acid, on PRRSV infection ability.
Methods
The cytotoxicity of DG was measured by MTT assay, and the effects of DG on PRRSV N gene/protein were investigated using real-time PCR, western blot and immunofluorescence assay. In addition, the effect of DG on cell apoptosis was analysed by fluorescence staining.
Results
Our results indicated that DG could effectively inhibit virus replication and N gene expression in MARC-145 cells infected with PRRSV. When the infected cells received DG, the numbers of apoptotic cells were decreased, and the cleaved caspase-3 contents were decreased dramatically.
Conclusions
Our study demonstrates that DG could effectively inhibit the PRRS virus via multiple pathways including inhibition of virus replication and N gene expression and reduction of apoptotic cells. DG can serve as a potential chemical for PRRSV prevention and control.
References
1.
RossowK.D.Porcine reproductive and respiratory syndrome. Vet Pathol1998; 35: 1–20.
2.
ZimmermanJ.J., YoonK.J., WillsR., SwensonS.General overview of PRRSV: a perspective from the United States. Vet Microbiol1997; 55: 187–196.
3.
DarwichL., DiazI., MateuE.Certainties, doubts and hypotheses in porcine reproductive and respiratory syndrome virus immunobiology. Virus Res2010; 154: 123–132.
4.
ChandR.J., TribleB.R., RowlandR.R.Pathogenesis of porcine reproductive and respiratory syndrome virus. Curr Opin Virol2012; 2: 256–263.
5.
MengX.J.Heterogeneity of porcine reproductive and respiratory syndrome virus: implications for current vaccine efficacy and future vaccine development. Vet Microbiol2000; 74: 309–329.
6.
OpriessnigT., HalburP., YoonK.J.Comparison of molecular and biological characteristics of a modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine (ingelvac PRRS MLV), the parent strain of the vaccine (ATCC VR2332), ATCC VR2385, and two recent field isolates of PRRSV. J Virol2002; 76: 11837–11844.
7.
MurtaughM.P., GenzowM.Immunological solutions for treatment and prevention of porcine reproductive and respiratory syndrome (PRRS). Vaccine2011; 29: 8192–8204.
WangL.J., GengC.A., MaY.B.Synthesis, biological evaluation and structure-activity relationships of glycyrrhetinic acid derivatives as novel anti-hepatitis B virus agents. Bioorg Med Chem Lett2012; 22: 3473–3479.
PliasunovaO., Il'inaT., KiselevaI.The anti-HIV activity of glycyrrhizic acid penta-O-nicotinate. Vestn Ross Akad Med Nauk2004; 11: 42–46. Russian.
12.
ItoM., SatoA., HirabayashiK.Mechanism of inhibitory effect of glycyrrhizin on replication of human immunodeficiency virus (HIV). Antiviral Res1988; 10: 289–298.
13.
HattoriT., IkematsuS., KoitoA.Preliminary evidence for inhibitory effect of glycyrrhizin on HIV replication in patients with AIDS. Antiviral Res1989; 11: 255–261.
14.
IshidaT., MizushinaY., YagiS.Inhibitory effects of glycyrrhetinic Acid on DNA polymerase and inflammatory activities. Evid Based Complement Alternat Med2012; 2012: 650514.
15.
ReedL.J., MuenchH.A simple method of estimating fifty per cent endpoints. Am J Epidemiol1938; 27: 493–497.
16.
MosmannT.Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods1983; 65: 55–63.
17.
ChenM.Z., XieH., YangL., LiaoZ., YuJ.In vitro anti-influenza virus activities of sulfated polysaccharide fractions from Gracilaria lemaneiformis. Virol Sin2010; 25: 341–351.
18.
MannsM.P., McHutchisonJ.G., GordonS.C.Peginterferon alfa-2b plus ribavirin compared with interferon alfa-2b plus ribavirin for initial treatment of chronic hepatitis C: a randomised trial. Lancet2001; 358: 958–965.
CinatlJ., MorgensternB., BauerG., ChandraP., RabenauH., DoerrH.Glycyrrhizin, an active component of liquorice roots, and replication of SARS-associated coronavirus. Lancet2003; 361: 2045–2046.
26.
HoeverG., BaltinaL., MichaelisM.Antiviral activity of glycyrrhizic acid derivatives against SARS-coronavirus. J Med Chem2005; 48: 1256–1259.
HaradaS.The broad anti-viral agent glycyrrhizin directly modulates the fluidity of plasma membrane and HIV-1 envelope. Biochem J2005; 392: 191.
29.
YooD., WoottonS.K., LiG., SongC., RowlandR.R.Colocalization and interaction of the porcine arterivirus nucleocapsid protein with the small nucleolar RNA-associated protein fibrillarin. J Virol2003; 77: 12173–12183.
30.
RowlandR.R.R., SchneiderP., FangY., WoottonS., YooD., BenfieldD.A.Peptide domains involved in the localization of the porcine reproductive and respiratory syndrome virus nucleocapsid protein to the nucleolus. Virology2003; 316: 135–145.
31.
SongC., LuR., BienzleD., LiuH.C., YooD.Interaction of the porcine reproductive and respiratory syndrome virus nucleocapsid protein with the inhibitor of MyoD family-a domain-containing protein. Biol Chem2009; 390: 215–223.
32.
SunY., HanM., KimC., CalvertJ.G., YooD.Interplay between interferon-mediated innate immunity and porcine reproductive and respiratory syndrome virus. Viruses2012; 4: 424–446.
33.
YangQ., GaoL., SiJ.Inhibition of porcine reproductive and respiratory syndrome virus replication by flavaspidic acid AB. Antiviral Res2013; 97: 66–73.
34.
WangX., BaiJ., ZhangL., LiY., JiangP.Poly(A)-binding protein interacts with the nucleocapsid protein of porcine reproductive and respiratory syndrome virus and participates in viral replication. Antiviral Res2012; 96: 315–323.
35.
BeuraL.K., DinhP.X., OsorioF.A., PattnaikA.K.Cellular poly(c) binding proteins 1 and 2 interact with porcine reproductive and respiratory syndrome virus nonstructural protein 1beta and support viral replication. J Virol2011; 85: 12939–12949.
36.
MillerL.C., FoxJ.M.Apoptosis and porcine reproductive and respiratory syndrome virus. Vet Immunol Immunopathol2004; 102: 131–142.
37.
ChoiC., ChaeC.Expression of tumour necrosis factor-α is associated with apoptosis in lungs of pigs experimentally infected with porcine reproductive and respiratory syndrome virus. Res Vet Sci2002; 72: 45–49.
38.
LeeS.M., KleiboekerS.B.Porcine reproductive and respiratory syndrome virus induces apoptosis through a mitochondria-mediated pathway. Virology2007; 365: 419–434.
39.
SuiX., YinJ., RenX.Antiviral effect of diammonium glycyrrhizinate and lithium chloride on cell infection by pseudorabies herpesvirus. Antiviral Res2010; 85: 346–353.
