title: Anti-inflammatory effect of garlic 14-kDa protein on LPS-stimulated-J774A.1 macrophages.
abstract: BACKGROUND
  Garlic 14-kDa protein is purified from garlic (Allium sativum L.) which is used in traditional medicine and exerts various immunomodulatory activities.
  
  
  OBJECTIVE
  The present study investigated the suppressive effect of garlic 14-kDa protein on LPS-induced expression of pro-inflammatory mediators and underlying mechanism in inflammatory macrophages.
  
  
  METHODS
  J774A.1 macrophages were treated with 14-kDa protein (5-30 μg/ml) with/without LPS (1 μg/ml) and the production of inflammatory mediators such as prostaglandin E2 (PGE2), TNF-α, and IL-1β released were measured using ELISA. Nitric oxide (NO) production was determined using the Griess method. The anti-inflammatory activity of 14-kDa protein was examined by measuring inducible nitric oxide synthase and cyclooxygenase-2 proteins using western blot. The expression of nuclear NF-κB p65 subunit was assessed by western blot.
  
  
  RESULTS
  Garlic 14-kDa protein significantly inhibited the excessive production of NO, PGE, TNF-α, and IL-1β in lipopolysaccharide (LPS)-activated J774A.1 macrophages in a concentration-related manner without cytotoxic effect. Western blot analysis demonstrated that garlic 14-kDa protein suppressed corresponding inducible NO synthase expression and activated cyclooxygenase-2 protein expression. The inhibitory effect was mediated partly by a reduction in the activity and expression of transcription factor NF-κB protein.
  
  
  CONCLUSIONS
  Our results suggested, for the first time, garlic 14-kDa protein exhibits anti-inflammatory properties in macrophages possibly by suppressing the inflammatory mediators via the inhibition of transcription factor NF-κB signaling pathway. The traditional use of garlic as anti-inflammatory remedy could be ascribed partly to 14-kDa protein content. This protein might be a useful candidate for controlling inflammatory diseases and further investigations in vivo.
authors.affiliation: Immunology Research Center, Bu-Ali Research Institute, School of Medicine, Mashhad University of Medical Sciences , Mashhad , Iran .
authors.forename: Shahrzad Zamani Taghizadeh
authors.initials: SZ
authors.lastname: Rabe
authors.forename: Tooba
authors.initials: T
authors.lastname: Ghazanfari
authors.forename: Zahra
authors.initials: Z
authors.lastname: Siadat
authors.forename: Maryam
authors.initials: M
authors.lastname: Rastin
authors.forename: Shahin Zamani Taghizadeh
authors.initials: SZ
authors.lastname: Rabe
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Mahmoudi
mesh_terms: D000893:Anti-Inflammatory Agents / Q000302:isolation & purification / Q000494:pharmacology*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D005737:Garlic; D006801:Humans; D018836:Inflammation Mediators / Q000037:antagonists & inhibitors / Q000276:immunology; D008070:Lipopolysaccharides / Q000633:toxicity*; D008264:Macrophages / Q000187:drug effects / Q000276:immunology*; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000276:immunology; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D010940:Plant Proteins / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D000893:Anti-Inflammatory Agents; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D016328:NF-kappa B; D010936:Plant Extracts; D010940:Plant Proteins
keywords: Allium sativum L.; J77A.1 macrophages; cyclooxygenase-2; inducible nitric oxide synthase; lipopolysaccharide