Opposing effect of p38 CCDPK and p44/42 CCDPK signaling on TNF-alpha-induced apoptosis in bovine aortic endothelial cells.
Acta Pharmacol Sin
; 22(5): 405-10, 2001 May.
Article
em En
| MEDLINE
| ID: mdl-11743886
ABSTRACT
AIM:
To investigate the pro-apoptotic role of tumor necrosis factor alpha (TNF-alpha) in cultured bovine aortic endothelial cells (BAEC) and its underlied apoptotic signaling pathways.METHODS:
BAEC were cultured and passaged in Dulbecco's modified Eagle's medium (DMEM). Morphologic changes and quantification of apoptotic cells were determined under fluorescence microscope after TNF-alpha treated BAEC for 24 h with Hoechst 33258 staining. Cell viability was determined with MTT method. DNA fragmentation was visualized by agarose gel electrophoresis. The expression of phospho-p38 and phospho-p44/42 Ca2+-calmodulin dependent protein kinase (CCDPK, formerly called MAPK) was measured by Western blotting.RESULTS:
TNF-alpha elicited typical apoptotic morphologic changes (chromatic condensation, nucleus fragmentation) and DNA fragmentation. At 1000-5000 kU/L, incubation with TNF-alpha for 24 h induced BAEC apoptosis and both of phospho-p38 and phospho-p44/42 CCDPK expression in a concentration-dependent manner. Interestingly, TNF-alpha-stimulated activation of p44/42 CCDPK was completely blocked, TNF-alpha-induced apoptosis was markedly increased by preincubation with U0126, a specific p44/42 CCDPK inhibitor. However, SB203580, a specific p38 CCDPK inhibitor, completely blocked TNF-alpha-stimulated activation of p38 CCDPK, and enhanced the expression of phospho-p44/42 CCDPK induced by TNF-alpha, substantially inhibited the pro-apoptotic effect of TNF-alpha.CONCLUSION:
TNF-alpha simultaneously activates p38 CCDPK and p44/42 CCDPK, and these two CCDPK signaling pathways appeared to play opposing roles in TNF-alpha-induced apoptosis in BAEC.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Endotélio Vascular
/
Fator de Necrose Tumoral alfa
/
Apoptose
/
Proteína Quinase 1 Ativada por Mitógeno
/
Proteínas Quinases Ativadas por Mitógeno
Limite:
Animals
Idioma:
En
Revista:
Acta Pharmacol Sin
Assunto da revista:
FARMACOLOGIA
Ano de publicação:
2001
Tipo de documento:
Article
País de afiliação:
China