RESUMO
The suitability of rabbit prothrombin activation fragment F 1.2 as a marker for the activation of the coagulation system was tested. Monoclonal antibodies to rabbit F 1.2 were raised, and a competitive F 1.2 ELISA was developed. Within the detection limit of the ELISA, no increase in rabbit F 1.2 was detected upon recalcification of plasma, whereas human F 1.2 increased 1500-fold. The apparent lack of F 1.2 formation in rabbit serum was confirmed by immunoblotting analysis of endogenous and biotin-labeled prothrombin. Meizothrombin and the B-chain of thrombin were the only prothrombin fragments detectable. In contrast, labeled human prothrombin formed, in addition, prethrombin 2 and F 1.2 in both human and rabbit serum. In contrast, rabbit F 1.2 formation could be demonstrated using purified rabbit prothrombin and factor Xa. These observations raise the possibility that rabbit prothrombin is less susceptible than the human counterpart to factor Xa cleavage at the 271/272 peptide bond. Thus, the primary structure of rabbit prothrombin was deduced by cDNA sequencing. While the 320/321 Xa cleavage site giving rise to meizothrombin was identical in rabbit and human prothrombin, the flanking region of the 271/272 Xa sensitive site contained a six amino acid deletion in the rabbit sequence. Taken together, these observations suggest that the observed differences between human and rabbit prothrombin activation may be due to different susceptibilities of the two Xa cleavage sites rather than plasma or serum cofactor(s).
Assuntos
Coagulação Sanguínea , Fragmentos de Peptídeos/fisiologia , Protrombina/fisiologia , Sequência de Aminoácidos , Animais , DNA Complementar/genética , Ensaio de Imunoadsorção Enzimática , Humanos , Dados de Sequência Molecular , Coelhos , Alinhamento de Sequência , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
We have examined the in vivo pharmacology of DuP 714 (Ac-[D]-Phe-Pro-boroArginine), a representative of a new series of synthetic thrombin inhibitors which contain a boronic acid derivative of arginine. Intravenous bolus injections of DuP 714 in anesthetized rats and conscious rabbits produced transient elevations of clotting times. Clinically relevant prolongations of the APTT were also observed in rabbits after i.v. infusion of less than 0.1 mg kg-1 h-1. Efficacy against venous thrombosis was demonstrated in a rabbit model of stasis induced thrombosis. Clots formed in 100% of control animals and only 33% of animals treated with 0.5 mg/kg DuP 714, and were less severe in treated animals. In a rabbit arterial-venous shunt model mimicking arterial thrombosis, occlusion occurred within 30 min in 72% of control animals vs. 11% of animals treated with 0.1 mg kg-1 h-1 DuP 714. Results indicate that DuP 714 is a highly effective anticoagulant which should be useful for the prevention of both venous and arterial thrombotic diseases.
Assuntos
Compostos de Boro/farmacologia , Oligopeptídeos/farmacologia , Trombina/antagonistas & inibidores , Animais , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Fibrinolíticos/farmacologia , Masculino , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Coelhos , Ratos , Ratos Endogâmicos , Tempo de Trombina , Tromboflebite/prevenção & controle , Trombose/prevenção & controleRESUMO
Based on data from a variety of experiments from several laboratories, adenosine appears to play an important role in the adjustment of blood flow to the metabolic requirements of the tissue. This has been shown to be true for heart, brain, and skeletal muscle in several different species. A reduction in oxygen supply or an increase in oxygen demand results in vasodilation and adenosine release. However, adenosine is also coupled to blood flow increments with enhanced metabolic activity and in the presence of an adequate oxygen supply. To what extent other vasoactive agents participate with adenosine in producing vasodilation under a variety of conditions is not known.