Determination of specific receptor sites for platelet activating factor in bovine neutrophils.
Am J Vet Res
; 65(5): 628-36, 2004 May.
Article
em En
| MEDLINE
| ID: mdl-15141884
OBJECTIVE: To identify and characterize a platelet activating factor (PAF) receptor in bovine neutrophils by use of radioligand binding, reverse transcription-polymerase chain reaction (RT-PCR) assay, and western blot analysis. ANIMALS: 4 healthy adult cows. PROCEDURE: Bovine neutrophil membranes were isolated for association, dissociation, and saturation binding experiments with PAF labeled with hydrogen 3 (3H-PAF). The RT-PCR assay was performed with appropriate human primers, and western blot analysis was developed with a polyclonal antibody obtained from a peptide of bovine PAF receptor. RESULTS: Analysis of kinetic binding data supported a single class of PAF receptor. Binding of 3H-PAF to membrane preparations was selectively displaced by PAF and a nonhydrolyzable analogue of guanine triphosphate (Gpp[NH]p) and by lyso-PAF (a biologically inactive analogue of PAF) to a lesser extent. Among other PAF receptor antagonists, 14-deoxyandrographolide and WEB 2086 were the most effective in inhibiting 3H-PAF binding sites in neutrophil membranes; 2 lignans, schisandrin-A and gamma-schisandrin were also effective, but 2 gingkolides (BN52020 and BN52021) only mildly inhibited 3H-PAF binding. Results of RT-PCR assay and western blot analysis of neutrophil crude membranes confirmed the presence of a PAF receptor. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that bovine neutrophils express only 1 type of PAF receptor, and it is likely that this receptor is involved in inflammatory responses. The most effective PAF antagonists were 14-deoxyandrographolide and WEB 2086; these PAF antagonists may be potentially useful in the treatment of inflammatory processes in cattle.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fator de Ativação de Plaquetas
/
Glicoproteínas da Membrana de Plaquetas
/
Receptores Acoplados a Proteínas G
/
Neutrófilos
Tipo de estudo:
Prognostic_studies
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Am J Vet Res
Ano de publicação:
2004
Tipo de documento:
Article
País de afiliação:
Chile
País de publicação:
Estados Unidos