NO自由基对猪圆环病毒2型复制的体外抑制作用研究

doi:10.16431/j.cnki.1671-7236.2019.03.034

Abstract

Abstract:

This study was conducted to investigate the inhibitory effect of NO· on the replication of porcine circovirus type 2 (PCV2).NO was generated from SNP or SNP in the presence of ascorbate (VC).SNP＋VC generates mainly NO· in culture medium while NO⁺ was the major product of SNP alone.PK-15 cells were pretreated with SNP,VC,SNP＋VC or control drug (NAP) for 6 h,respectively,then infected with PCV2 (1 MOI) and incubated with the above mentioned drugs for additional 72 h.After incubation,culture medium and cells were collected.IFA,Western blotting,TCID₅₀ and QPCR assay methods were performed to determine the viral titers.The results showed that the highest concentration of SNP,VC and SNP＋VC in treated cells were 500,62.5 and 31.25 μmol/L,respectively.The production of NO in 60 μmol/L SNP or 30 μmol/L SNP＋30 μmol/L VC treated cells were higher than that in untreated cells and NAP treated cells (P<0.01).Virus titers and PCV2 DNA copies from 30 μmol/L SNP＋30 μmol/L VC treated group were significantly lower than that from infected control (P<0.01).In addition,The expression of viral Cap protein from the sample treated with 30 μmol/L SNP＋30 μmol/L VC also obviously decreased in relation to those from infected control samples.However,almost no antiviral activities of SNP or VC alone were indicated during PCV2 infection.In conclusion,inhibition of NO on PCV2 replication was mediated by NO·,rather than NO⁺.

Key words: NO; NO·; porcine circovirus type 2 (PCV2); inhibition

CLC Number:

S852.65

LI Jizong, ZHANG Dongxia, LI Li, XUE Tao, LIU Chuanmin. The Free Radical of Nitric Oxide Inhibits Porcine Circovirus Type 2 Replicion in vitro[J]. , 2019, 46(3): 924-930.

References

[1] TISCHER I,RASCH R,TOCHTERMANN G.Characterization of papovavirus-and picornavirus-like particles in permanent pig kidney cell lines[J].Zentralbl Bakteriol Orig A,1974,226(2):153-167.
[2] LI J,SHI J L,WU X Y,et al.Differentiation of PCV1 and PCV2 by a multiplex Real-time PCR assay[J].Veterinary Record,2013,173(14):346.
[3] OPRIESSNIG T,LANGOHR I.Current state of knowledge on porcine circovirus type 2-associated lesions[J].Veterinary Pathology,2013,50(1):23-38.
[4] LLORENÇ G R,ANDERS S,CHARLOTTE S K,et al.Infectious risk factors for individual postweaning multisystemic wasting syndrome (PMWS) development in pigs from affected farms in Spain and Denmark[J].Research in Veterinary Science,2012,93(3):1231-1240.
[5] MARTINS G D,CASTRO A M,CORTEZ A,et al.Genetic diversity of Brazilian strains of porcine circovirus type 2(PCV-2) revealed by analysis of the cap gene (ORF-2)[J].Archives of Virology,2007,152(8):1435-1445.
[6] BOGDAN C,RÖLLINGHOFF M,DIEFENBACH A.Reactive oxygen and reactive nitrogen intermediates in innate and specific immunity[J].Current Opinion in Immunology,2000,12(1):64-76.
[7] BOGDAN C,RÖLLINGHOFF M,DIEFENBACH A.The role of nitric oxide in innate immunity[J].Immunological Reviews,2000,173(1):17-26.
[8] 陈耿,张书霞,华利忠.PCV2对仔猪胸腺IL-2、IL-6和IL-10及其受体mRNA的影响及NO-NOS系统的调控[J].南京农业大学学报,2011,34(3):101-106. CHEN G,ZHANG S X,HUA L Z.Effect of PCV2 on mRNA of IL-2,IL-6,IL-10 and their receptors in piglet thymus and the regulation of NO-NOS system[J].Journal of Nanjing Agricultural University,2011,34(3):101-106.(in Chinese)
[9] REN W K,YIN Y L,YU X L,et al.Effect of dietary arginine supplementation on reproductive performance of mice with porcine circovirus type 2 infection[J].Amino Acids,2012,42(6):2089.
[10] LIU G,REN W K,SU D D,et al.Dietary L-arginine supplementation improves the immune responses in mouse model infected porcine circovirus types 2[J].Journal of Animal and Veterinary Advances,2012,11(16):2980-2985.
[11] REN W K,LIU G,LI T J,et al.Dietary supplementation with arginine and glutamine confers a positive effect in porcine circovirus-infected pig[J].Journal of Food Agriculture & Environment,2012,10(2):485-490.
[12] SUN J,ZHANG X,BRODERICK M,et al.Measurement of nitric oxide production in biological systems by using griess reaction assay[J].Sensors,2003,3(8):276-284.
[13] REED L J,MUENCH H.A simple method of estimation of fifty percent endpoint[J].American Journal of Epidemiology,1938,27(3):493-497.
[14] OPRIESSNIG T,YU S,GALLUP J M,et al.Effect of vaccination with selective bacterins on conventional pigs infected with type 2 porcine circovirus[J].Veterinary Pathology,2003,40(5):521-529.
[15] SAURA M,ZARAGOZA C,MCMILLAN A,et al.An antiviral mechanism of nitric oxide:Inhibition of a viral protease[J].Immunity,1999,10(1):21-28.
[16] LIN Y L,HUANG Y L,MA S H,et al.Inhibition of Japanese encephalitis virus infection by nitric oxide:Antiviral effect of nitric oxide on RNA virus replication[J].Virology,1997,71(7):5227-5235.
[17] AKERSTRÖM S,GUNALAN V,KENG C T,et al.Dual effect of nitric oxide on SARS-CoV replication:Viral RNA production and palmitoylation of the S protein are affected[J].Virology,2009,395(1):1-9.
[18] AKAIKE T.Role of free radicals in viral pathogenesis and mutation[J].Reviews in Medical Virology,2001,11(2):87-101.
[19] THOMAS D D,RIDNOUR L A,ISENBERG J S,et al.The chemical biology of nitric oxide:Implications in cellular signaling[J].Free Radical Biology And Medcine,2008,45(1):18-31.
[20] SHERIDAN M P,REGEV-SHOSHANI G,MARTINS J,et al.Nitric oxide modulates the immunological response of bovine PBMCs in an in vitro BRDc infection model[J].Research in Veterinary Science,2016,109:21-28
[21] ANDERSON S M,NAIDOO R N,PILLAY Y,et al,HIV induced nitric oxide and lipid peroxidation,influences neonatal birthweight in a South African population[J].Environment International,2018,121(Pt 1):1-12.
[22] LIU C M,WEN L B,XIAO Q,et al.Nitric oxide-generating compound GSNO suppresses porcine circovirus type 2 infection in vitro and in vivo[J].BMC Veterinary Research,2017,13(1):59.
[23] UBOL S,HIRIOTE W,ANUNTAGOOL N,et al.A radical form of nitric oxide suppresses RNA synthesis of rabies virus[J]. Virus Research,2001,81(1-2):125-132.

The Free Radical of Nitric Oxide Inhibits Porcine Circovirus Type 2 Replicion in vitro

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