title: Effects of sirtuin 1 activation on nicotine and lipopolysaccharide-induced cytotoxicity and inflammatory cytokine production in human gingival fibroblasts.
abstract: OBJECTIVE
  Although sirtuin 1 (SIRT1) over-expression and resveratrol exert anti-inflammatory and proinflammatory effects, their effects and the mechanism of action on human gingival fibroblast (HGF)-mediated inflammation are unknown. The aim of this study was to demonstrate the effects of activating SIRT1 using resveratrol and recombinant adenovirus encoding SIRT1 (Ad-SIRT1) on the expression of proinflammatory cytokines and to elucidate its mechanism of action of lipopolysaccharide (LPS) and nicotine stimulated-HGF.
  
  
  METHODS
  Cytotoxicity and the production of reactive oxygen species (ROS) were measured using the 3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively. The amount of prostaglandin E2 (PGE2 ) released into the culture medium was measured by radioimmunoassay. mRNA and protein levels were analyzed using RT-PCR and western blotting, respectively.
  
  
  RESULTS
  Nicotine and LPS up-regulated the expression of SIRT1 mRNA and SIRT1 protein in a time- and concentration-dependent manner. Resveratrol and Ad-SIRT1 decreased LPS and nicotine-induced cytotoxicity, ROS and PGE2 production, and expression of cyclooxygenase-2 in HGFs. Resveratrol and Ad-SIRT1 inhibited nicotine and LPS-mediated protein kinase C (PKC), phosphatidylinositol 3-kinase (PI3K), p38, ERK, JNK, MAPK and nuclear factor-kappa B (NF-κB) activation.
  
  
  CONCLUSIONS
  This study is the first to show that the anti-inflammatory and cytoprotective effects of SIRT1 activation in HGFs occur through the PKC, PI3K, MAPK and NF-κB pathways.
authors.affiliation: Department of Oral Histology, School of Dentistry, Dankook University, Cheon-An, Korea.
authors.forename: G-J
authors.initials: GJ
authors.lastname: Park
authors.forename: Y-S
authors.initials: YS
authors.lastname: Kim
authors.forename: K-L
authors.initials: KL
authors.lastname: Kang
authors.forename: S-J
authors.initials: SJ
authors.lastname: Bae
authors.forename: H-S
authors.initials: HS
authors.lastname: Baek
authors.forename: Q-S
authors.initials: QS
authors.lastname: Auh
authors.forename: Y-H
authors.initials: YH
authors.lastname: Chun
authors.forename: B-H
authors.initials: BH
authors.lastname: Park
authors.forename: E-C
authors.initials: EC
authors.lastname: Kim
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D051546:Cyclooxygenase 2 / Q000187:drug effects; D015232:Dinoprostone / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects*; D005822:Genetic Vectors / Q000235:genetics; D005881:Gingiva / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D007378:Interleukins / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D048670:MAP Kinase Kinase 4 / Q000037:antagonists & inhibitors; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D009538:Nicotine / Q000494:pharmacology*; D018722:Nicotinic Agonists / Q000494:pharmacology*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D017382:Reactive Oxygen Species / Q000378:metabolism; D000077185:Resveratrol; D056564:Sirtuin 1 / Q000235:genetics / Q000494:pharmacology*; D013267:Stilbenes / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects; D015854:Up-Regulation; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D018836:Inflammation Mediators; D007378:Interleukins; D008070:Lipopolysaccharides; D016328:NF-kappa B; D018722:Nicotinic Agonists; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017382:Reactive Oxygen Species; D013267:Stilbenes; D014409:Tumor Necrosis Factor-alpha; D009538:Nicotine; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4; C447939:SIRT1 protein, human; D056564:Sirtuin 1; D015232:Dinoprostone; D000077185:Resveratrol