Contribution of toxins isolated from Bothrops alternatus (yarara de la cruz) venom on the myotoxicity of C2C12 cell line.

GARCIA DENEGRI M.E.; BUSTILLO S.; GAY C.; GORODNER J.O.; PONCE-SOTO L.A.; LEIVA L.; ACOSTA O.

Huerta Grande, Córdoba.

Congreso; Cordoba Biology Society. XVII Annual Scientific Meeting.; 2009

Sociedad de Biologia de Cordoba.

Previous in vivo and in vitro assays demonstrated the myotoxic activity from B. alternatus crude venom. The aim of this work was to evaluate the contribution of a phospholipase A2 (PLA2) and a metalloproteinase (baltergin) to the in vitro myotoxicity in murine myoblast cell culture. Both toxins were purified from B. alternatus venom. Then, the cytotoxic activity displayed by each enzyme and also of the whole venom pre-incubated with anti-baltergin (a-balt) in different dilutions was studied. Cells (into DMEM - 5% of SFB - 5% CO2, 37° C) were treated with different concentrations of the isolated enzymes (up to 200 µg/ml) and crude venom (200 µg/ml) pre-incubated with different amounts of a-balt. The cell viability was measured, after 3 h of exposure, by colorimetric method (crystal violet staining). Both the PLA2 and the venom pre-incubated with a-balt do not show myotoxic activity, whereas baltergine reduced the cell viability by about 30 %. Due to the exposure of cells to the whole venom (100 µg/mL) produced 100% cytotoxicity and the a-balt antibodies neutralized completely the myotoxic activity showed by the venom, we can conclude that the metalloproteinase is capable, by itself, of damage on muscular fibers. In spite of the lack of in vitro myotoxic activity by PLA2, a synergic action of both enzymes could develop an acute muscle lesion characteristic for B. alternatus intoxications.