title: Acetylsalicylic acid inhibits hepatitis C virus RNA and protein expression through cyclooxygenase 2 signaling pathways.
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.
  
  
  CONCLUSIONS
  These findings suggest the possibility that ASA could be an excellent adjuvant in the treatment of chronic HCV infection.
authors.affiliation: Department of Biochemistry, School of Medicine and University Hospital, Autonomous University of Nuevo Leon, Monterrey, Nuevo Leon, Mexico.
authors.forename: Karina
authors.initials: K
authors.lastname: Trujillo-Murillo
authors.forename: Ana Rosa
authors.initials: AR
authors.lastname: Rincón-Sánchez
authors.forename: Herminia
authors.initials: H
authors.lastname: Martínez-Rodríguez
authors.forename: Francisco
authors.initials: F
authors.lastname: Bosques-Padilla
authors.forename: Javier
authors.initials: J
authors.lastname: Ramos-Jiménez
authors.forename: Hugo A
authors.initials: HA
authors.lastname: Barrera-Saldaña
authors.forename: Marcos
authors.initials: M
authors.lastname: Rojkind
authors.forename: Ana María
authors.initials: AM
authors.lastname: Rivas-Estilla
mesh_terms: D001241:Aspirin / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000502:physiology*; D005838:Genotype; D016174:Hepacivirus / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D008113:Liver Neoplasms; D008156:Luciferases / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D012367:RNA, Viral / Q000187:drug effects / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects*; D014162:Transfection; D014764:Viral Proteins / Q000187:drug effects / Q000235:genetics*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D012334:RNA, Neoplasm; D012367:RNA, Viral; D014764:Viral Proteins; D008156:Luciferases; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D001241:Aspirin