Acetylsalicylic acid inhibits hepatitis C virus RNA and... : Hepatology (original) (raw)

Viral Hepatitis

Acetylsalicylic acid inhibits hepatitis C virus RNA and protein expression through cyclooxygenase 2 signaling pathways

Trujillo-Murillo, Karina1; Rincón-Sánchez, Ana Rosa4; Martínez-Rodríguez, Herminia1; Bosques-Padilla, Francisco2; Ramos-Jiménez, Javier3; Barrera-Saldaña, Hugo A.1; Rojkind, Marcos5; Rivas-Estilla, Ana María1*

1_Department of Biochemistry, School of Medicine and University Hospital, Autonomous University of Nuevo Leon, Monterrey, Nuevo Leon, Mexico_

2_Department of Gastroenterology, School of Medicine and University Hospital, Autonomous University of Nuevo Leon, Monterrey, Nuevo Leon, Mexico_

3_Department of Infectology, School of Medicine and University Hospital, Autonomous University of Nuevo Leon, Monterrey, Nuevo Leon, Mexico_

4_University Center of Health Sciences, University of Guadalajara, Jalisco, Mexico_

5_Department of Biochemistry and Molecular Biology, George Washington, University Medical Center, Washington, DC_

*Address reprint requests to: Department of Biochemistry, School of Medicine, Autonomous University of Nuevo Leon, Monterrey, Nuevo Leon, Mexico

Email:[email protected]

Received 16 August 2007; Accepted 2 January 2008

Published online in Wiley InterScience (www.interscience.wiley.com).

Potential conflict of interest: Nothing to report.

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Abstract

It has been reported that salicylates (sodium salicylate and aspirin) inhibit the replication of flaviviruses, such as Japanese encephalitis virus and dengue virus. Therefore, we considered it important to test whether acetylsalicylic acid (ASA) had anti–hepatitis C virus (HCV) activity. To this end, we examined the effects of ASA on viral replication and protein expression, using an HCV subgenomic replicon cell culture system. We incubated Huh7 replicon cells with 2-8 mM ASA for different times and measured HCV-RNA and protein levels by northern blot, real-time polymerase chain reaction, and western analysis, respectively. We found that ASA had a suppressive effect on HCV-RNA and protein levels (nearly 58%). ASA-dependent inhibition of HCV expression was not mediated by the 5′-internal ribosome entry site or 3′-untranslated regions, as determined by transfection assays using bicistronic constructs containing these regulatory regions. However, we found that HCV-induced cyclooxygenase 2 (COX-2) messenger RNA and protein levels and activity and these effects were down-regulated by ASA, possibly by a nuclear factor kappa B–independent mechanism. We also observed that the ASA-dependent inhibition of viral replication was due in part to inhibition of COX-2 and activation of p38 and mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1/2 (MEK1/2) mitogen-activated protein kinases (MAPKs). Inhibition of these kinases by SB203580 and U0126, respectively, and by short interfering RNA silencing of p38 and MEK1 MAPK prevented the antiviral effect of ASA. Taken together, our findings suggest that the anti-HCV effect of ASA in the Huh7 replicon cells is due to its inhibitory effect on COX-2 expression, which is mediated in part by the activation of MEK1/2/p38 MAPK. Conclusion: These findings suggest the possibility that ASA could be an excellent adjuvant in the treatment of chronic HCV infection.

Abbreviations: ASA, acetylsalicylic acid; bp, base pair; cDNA, complementary DNA; COX, cyclooxygenase; DENV, dengue virus; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HCV, hepatitis C virus; IRES, internal ribosome entry site; JEV, Japanese encephalitis virus; JNK, c-Jun N-terminal kinase; MAb, monoclonal antibody; MAPK, mitogen-activated protein kinase; MEK, mitogen-activated protein kinase/extracellular signal-regulated kinase kinase; mRNA, messenger RNA; NF-κB, nuclear factor kappa B; NPT II, neomycin phosphotransferase II; NS5A, nonstructural protein 5A; NSAID, nonsteroidal anti-inflammatory drug; PCR, polymerase chain reaction; PGE2, prostaglandin E2; RLU, relative luciferase unit; RT-PCR, reverse-transcription polymerase chain reaction; siRNA, short interfering RNA; UTR, untranslated region.