Resumo

GARCIA DENEGRI, M.E. et al. Actividades farmacológicas de la fosfolipasa A₂ Ba SpII RP4 del veneno de Bothrops alternatus. Rev. vet. [online]. 2012, vol.23, n.1, pp.25-31. ISSN 1669-6840.

The aim of this work was to evaluate the pharmacological activities (citotoxicity, bactericidal, platelet aggregation) and the changes in the activity/stability of the isolated phospholipase A2 (PLA2) from Bothrops alternatus venom. In order to elucidate myotoxic and bactericidal activities, murine mioblast /miotubules (C2C12) and strains of Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) were used, respectively. Also, normal mammary epithelial cell culture (NMuMG) and tumoral type (LM3) were tested for a possible role in oncological therapy. With the purpose of studying the inhibitory effect of the PLA2 on platelet aggregation, washed human platelet and thrombin as physiological inductor, were used. Finally, changes in the activity and stability of the isolated PLA2 at different temperatures (4-90ºC) by kinetic assays were recorded; while the enzyme was subjected to radial indirect hemolysis to evaluate the effect of different pH (2.5-10). The PLA2 presented dose-dependent inhibitory effect on thrombin-induced platelet aggregation. Also, it showed high structural stability when exposed to extreme conditions of temperature and pH during several hours, and catalytic activity remained almost unchanged. Nevertheless, this enzyme tested on different cells cultures did not induce cytotoxic (even at high doses) or bactericidal activities. These findings have a scientific value for toxicology since they expand the knowledge of the characterization (biological, biochemical and structural aspects) of one of the most abundant enzyme (PLA2 Ba SpII RP4) from the protein package of B. alternatus snake venom from the northeast of Argentina. Likewise, the confirmed safety and stability of this PLA2 strongly suggest that it could be proposed as an alternative immunogen for the antiserum production against Bothrops snake venom

Palavras-chave : Bothrops alternatus; phospholipase A2; bactericidal activity; citotoxicity; platelet aggregation; immunogenicity.

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