title: Effect of dibutyryl cyclic adenosine monophosphate on the gene expression of plasminogen activator inhibitor-1 and tissue factor in adipocytes.
abstract: BACKGROUND
  Hypertrophic adipocytes in obese states express the elevated levels of plasminogen activator inhibitor-1 (PAI-1) and tissue factor (TF). An increase in the intracellular concentration of cyclic adenosine monophosphate (cAMP) promotes triglyceride hydrolysis and may improve dysregulation of adipocyte metabolism. Here, we investigate the effect of dibutyryl-cAMP (a phosphodiesterase-resistant analog of cAMP) on the gene expression of PAI-1 and TF in adipocytes.
  
  
  METHODS
  Differentiated 3T3-L1 adipocytes were treated with dibutyryl-cAMP and agents that would be expected to elevate intracellular cAMP, including cilostazol (a phosphodiesterase inhibitor with anti-platelet and vasodilatory properties), isoproterenol (a beta adrenergic agonist) and forskolin (an adenylyl cyclase activator). The levels of PAI-1 and TF mRNAs were measured using real-time quantitative reverse transcription-PCR.
  
  
  CONCLUSIONS
  The treatment of adipocytes with dibutyryl-cAMP resulted in the inhibition of both lipid accumulation and TF gene expression. However, PAI-1 gene expression was slightly but significantly increased by dibutyryl-cAMP. On the other hand, cilostazol inhibited the expression of PAI-1 without affecting lipid accumulation. When the adipocytes were treated with cilostazol in combination with isoproterenol or forskolin, the inhibitory effect of cilostazol on PAI-1 gene expression was counteracted, thus suggesting that inhibition by cilostazol may not be the result of intracellular cAMP accumulation by phosphodiesterase inhibition. These results suggest the implication of cAMP in regulation of the gene expression of TF and PAI-1 in adipocytes. Our findings will serve as a useful basis for further research in therapy for obesity-associated thrombosis.
authors.affiliation: Division of Pathological Biochemistry, Department of Life Sciences, Tottori University School of Medicine, Yonago 683–8503, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Taniguchi
authors.forename: Naoko
authors.initials: N
authors.lastname: Ono
authors.forename: Akira
authors.initials: A
authors.lastname: Hayashi
authors.forename: Yuwna
authors.initials: Y
authors.lastname: Yakura
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takeya
mesh_terms: D041721:3T3-L1 Cells; D000262:Adenylyl Cyclases / Q000378:metabolism; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology*; D000077407:Cilostazol; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D004789:Enzyme Activation; D020536:Enzyme Activators / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006984:Hypertrophy; D007545:Isoproterenol / Q000494:pharmacology; D050356:Lipid Metabolism / Q000187:drug effects; D051379:Mice; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013777:Tetrazoles / Q000494:pharmacology; D013925:Thromboplastin / Q000235:genetics*
chemical_list: D000318:Adrenergic beta-Agonists; D020536:Enzyme Activators; D010726:Phosphodiesterase Inhibitors; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D013777:Tetrazoles; D005576:Colforsin; D003994:Bucladesine; D013925:Thromboplastin; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D007545:Isoproterenol; D000077407:Cilostazol