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Downregulation of inducible nitric oxide synthase (iNOS) expression is implicated in the antiviral activity of acetylsalicylic acid in HCV-expressing cells

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Abstract

Previously, we described that acetylsalicylic acid (ASA) decreases HCV expression, but the mechanisms involved have not been clearly established. We evaluated the participation of inducible nitric oxide synthase (iNOS) in the regulation of HCV-RNA induced by ASA. Huh7 cells expressing non-structural HCV proteins were exposed to 4 mM ASA and incubated at the same times we reported HCV downregulation (24-72 h), and iNOS mRNA and protein levels were then measured by real-time PCR and Western blot, respectively. Nitric oxide levels were measured at the same time. Inhibition of iNOS mRNA by small interfering RNAs (siRNA) and activation of the iNOS gene promoter by ASA treatment were evaluated. In Huh7 replicon cells treated with ASA, we found decreased levels of iNOS mRNA, iNOS protein and nitrosylated protein levels at 48-72 h. ASA exposure also reduced the transactivation of the iNOS promoter in HCV replicon cells at 48 h, and this was partly due to the decrease in the affinity of transcription factor C/EBP-β for its binding site in the iNOS promoter. siRNA silencing of iNOS decreased HCV-RNA expression (65 %) and potentiated the antiviral effect (80 %) of ASA compared with control cells. ASA reduces iNOS expression by downregulating promoter activity, mRNA and protein levels at the same time that it decreases HCV expression. These findings suggest that the antiviral activity of ASA is mediated partially through the modulation of iNOS.

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Abbreviations

ASA:

Acetylsalicylic acid

HCV:

Hepatitis C virus

iNOS:

Inducible nitric oxide synthase

NO:

Nitric oxide

NSAIDs:

Nonsteroidal anti-inflammatory drugs

RT-PCR:

Reverse transcription polymerase chain reaction

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Acknowledgments

We thank Sergio Lozano-Rodriguez, M.D. (UANL) for his assistance in reviewing the manuscript. Financial support was provided by the CONACYT, grant number SALUD-2008-C01-86996, BASICA-CB2010-01-155082 and INFR-224638 awarded to AM Rivas.

Conflict of interest

No personal interest to declare.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Medicine, School of Medicine, Autonomous University of Nuevo Leon, Ave. Francisco I. Madero y Ave. Gonzalitos s/n CP 64460, Col. Mitras Centro, Monterrey, Nuevo León, Mexico

    Clara Patricia Ríos-Ibarra, Sonia Lozano-Sepulveda, Carlos Cordova-Fletes & Ana María G. Rivas-Estilla

  2. Unit Liver, Department of Internal Medicine, School of Medicine, Autonomous University of Nuevo León, Monterrey, Nuevo León, Mexico

    Linda Muñoz-Espinosa

  3. CUCS, University of Guadalajara, Jalisco, Mexico

    Ana Rosa Rincón-Sánchez

Authors
  1. Clara Patricia Ríos-Ibarra
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  2. Sonia Lozano-Sepulveda
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  3. Linda Muñoz-Espinosa
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  4. Ana Rosa Rincón-Sánchez
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  5. Carlos Cordova-Fletes
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  6. Ana María G. Rivas-Estilla
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Corresponding author

Correspondence to Ana María G. Rivas-Estilla.

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Ríos-Ibarra, C.P., Lozano-Sepulveda, S., Muñoz-Espinosa, L. et al. Downregulation of inducible nitric oxide synthase (iNOS) expression is implicated in the antiviral activity of acetylsalicylic acid in HCV-expressing cells. Arch Virol 159, 3321–3328 (2014). https://doi.org/10.1007/s00705-014-2201-5

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  • DOI: https://doi.org/10.1007/s00705-014-2201-5

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