title: KF19514, a phosphodiesterase 4 and 1 inhibitor, inhibits TNF-alpha-induced GM-CSF production by a human bronchial epithelial cell line via inhibition of PDE4.
abstract: OBJECTIVE
  Bronchial epithelium plays an important role in the regulation of inflammatory reactions in the airways. We investigated the effect of KF19514, a phosphodiesterase (PDE) 4 and 1 inhibitor, on granulocyte-macrophage colony-stimulating factor (GM-CSF) production by a human bronchial epithelial cell line, BEAS-2B.
  
  
  METHODS
  BEAS-2B cells were stimulated with the tumor necrosis factor-alpha (TNF-alpha) and various concentrations of test agents for 48 h. Supernatants were assayed for GM-CSF by using an enzyme-linked immunosorbent assay (ELISA). In addition, intracellular cyclic AMP (cAMP) levels were measured in the presence of various agents.
  
  
  RESULTS
  KF19514 significantly inhibited the release of GM-CSF by BEAS-2B cells in a concentration-dependent manner. The other PDE4 inhibitors and cAMP-elevating agents also inhibited the GM-CSF production. In the BEAS-2B cells, KF19514 and PDE4 inhibitors concentration-dependently increased intracellular cAMP levels. The inhibitory effect of KF19514 on the GM-CSF production was significantly reduced by a cAMP-dependent protein kinase A (PKA) inhibitor, H89. On the other hand, other PDE isozyme inhibitors did not inhibit the GM-CSF production by BEAS-2B cells, and did not elevate the intracellular cAMP levels.
  
  
  CONCLUSIONS
  These results indicate that KF19514 and PDE4 inhibitors reduce TNF-alpha-induced GM-CSF production of BEAS-2B cells via a cAMP-dependent pathway. PDE4 may be a possible target for the regulation of cytokine production in epithelial cells.
authors.affiliation: Pharmaceutical Research Institute, Kyowa Hakko Kogyo Co., Ltd., 1188 Shimotogari, Nagaizumi-cho, Sunto-gun, Shizuoka 411-8731, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sasaki
authors.forename: H
authors.initials: H
authors.lastname: Manabe
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000037:antagonists & inhibitors*; D001980:Bronchi / Q000187:drug effects / Q000378:metabolism*; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D007527:Isoenzymes / Q000037:antagonists & inhibitors; D007546:Isoquinolines / Q000494:pharmacology; D009287:Naphthyridines / Q000494:pharmacology*; D043264:Phosphodiesterase I / Q000037:antagonists & inhibitors; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D020545:Respiratory Mucosa / Q000187:drug effects / Q000378:metabolism; D013449:Sulfonamides; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D007527:Isoenzymes; D007546:Isoquinolines; C106878:KF 19514; D009287:Naphthyridines; D010726:Phosphodiesterase Inhibitors; D013449:Sulfonamides; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D043264:Phosphodiesterase I; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide