Pole of the cyclic AMP-protein kinase a pathway in lipopolysaccharide-induced nitric oxide synthase expression in RAW 264.7 macrophages - Involvement of cyclooxygenase-2

被引:108
作者
Chen, CC [1 ]
Chiu, KT [1 ]
Sun, YT [1 ]
Chen, WC [1 ]
机构
[1] Natl Taiwan Univ, Coll Med, Dept Pharmacol, Taipei 10018, Taiwan
关键词
D O I
10.1074/jbc.274.44.31559
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The signaling pathway for lipopolysaccharide (LPS)-induced nitric oxide (NO) release in RAW 264.7 macrophages involves the protein kinase C and p38 activation pathways (Chen, C. C., Wang, J. K., and Lin, S. B. (1998) J. Immunol. 161, 6206-6214; Chen, C. C., and Wang, J. K. (1999) Mel. Pharmacol. 55, 481-488). In this study, the role of the cAMP-dependent protein kinase A (PKA) pathway was investigated. The PEA inhibitors, KT-5720 and H8, reduced LPS-induced NO release and inducible nitric oxide synthase (iNOS) expression. The direct PHA activator, Bt(2)cAMP, caused concentration-dependent NO release and iNOS expression, as confirmed by immunofluorescence studies. The intracellular cAMP concentration did not increase until after 6 h of LPS treatment. Two cAMP-elevating agents, forskolin and cholera toxin, potentiated the LPS-induced NO release and iNOS expression. Stimulation of cells with LPS or Bt(2)cAMP for periods of 10 min to 24 h caused nuclear factor-kappa B (NF-kappa B) activation in the nuclei, as shown by detection of NF-kappa B-specific DNA-protein binding The PKA inhibitor, H8, inhibited the NF-kappa B activation induced by 6- or 12-h treatment with LPS but not that induced after 1, 3, or 24 h. The cyclooxygenase-a (COX-2) inhibitors, NS-398 and indomethacin, attenuated LPS-induced NO release, iNOS expression, and NF-kappa B DNA-protein complex formation. LPS induced COX-2 expression in a time-dependent manner, and prostaglandin E-2 production was induced in parallel. These results suggest that 6 h of treatment with LPS increases intracellular cAMP levels via COX-2 induction and prostaglandin E-2 production, resulting in PKA activation, NF-kappa B activation, iNOS expression, and NO production.
引用
收藏
页码:31559 / 31564
页数:6
相关论文
共 33 条
[1]  
Chen CC, 1999, MOL PHARMACOL, V55, P481
[2]  
Chen CC, 1998, J IMMUNOL, V161, P6206
[3]   YES, BUT DO THEY STILL GET HEADACHES [J].
DEWITT, D ;
SMITH, WL .
CELL, 1995, 83 (03) :345-348
[4]  
FOURNIER T, 1995, J IMMUNOL, V155, P2123
[5]  
FUJIHARA M, 1993, J BIOL CHEM, V268, P14898
[6]   INDUCTION OF NITRIC-OXIDE SYNTHASE IN CULTURED VASCULAR SMOOTH-MUSCLE CELLS - THE ROLE OF CYCLIC-AMP [J].
HIROKAWA, K ;
OSHAUGHNESSY, K ;
MOORE, K ;
RAMRAKHA, P ;
WILKINS, MR .
BRITISH JOURNAL OF PHARMACOLOGY, 1994, 112 (02) :396-402
[7]  
JEAN YJ, 1996, MOL PHARMACOL, V50, P334
[8]   On the induction of cyclooxygenase-2, inducible nitric oxide synthase and soluble phospholipase A2 in rat mesangial cells by a nonsteroidal anti-inflammatory drug:: The role of cyclic AMP [J].
Klein, T ;
Ullrich, V ;
Pfeilschifter, J ;
Nüsing, R .
MOLECULAR PHARMACOLOGY, 1998, 53 (03) :385-391
[9]   In murine 3T3 fibroblasts, different second messenger pathways resulting in the induction of NO synthase II (iNOS) converge in the activation of transcription factor NF-kappa B [J].
Kleinert, H ;
Euchenhofer, C ;
IhrigBiedert, I ;
Forstermann, U .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (11) :6039-6044
[10]  
KOIDE M, 1993, J BIOL CHEM, V268, P24959