Study of myotoxic activity of Bothrops leucurus venom (Serpentes; Viperidae) abd protective action of Callendula officinalis (Asteraceae; Compositae)

1Mise, Y.; ²Casais-e-Silva, L.L.; ³Barbosa Jr, A. and ^1,4Lira-da-Silva, R.M.

1 Depto. Zoologia, IB, UFBA, Bahia; 2 Escola Bahiana de Medicina e Saúde Pública, Bahia; 3 FIOCRUZ, Bahia, 4 Faculdade de Tecnologia e Ciências, Bahia

The main objectives of the present work are: a) verify the dose-response relation in the myotoxicity of Bothropsleucurus venom,b) measure the level of muscular damage in gastrocnemius muscle related to observed histopathological alterations, c)challenge the applicability of Callendulaofficinalis for previous and associated treatment in bothropic accidents. Miotoxicity was evaluated in vivo by the intramuscular inoculation of 50mL of B. leucurus crude venom (tested with different concentrations of 25, 50 or 100mg/mL). Control test was made injecting salina in the same muscle.The effects of C. officinalis, were evaluated in the different forms, with incubation and inoculation, intramuscular inoculation of venom and, 20 minutes after the inoculation, topical treatment and intraperitoneal injection. Orbital punction was done in order to extract blood samples, at 1,3,6,12 and 24 hours after the inoculation of the solution. The plasma was separated by centrifugation and took the upper fraction. The samples were kept at 4°C during all the process. The CK activity was determinated by CK-NAC liquiform kit.In order to make a histological analysis, the muscle was removed, fixed at formol 10%,and processed for optic microscopy. B.leucurusvenom showed miotoxic activity in vivo, characterized by release of CK and histopathological alterations in gastrocnemius muscle of mice. In the experiment, kinetic of CK released resulted temporal and dose-dependant. The maximun level reached in liberation of CK happens between 3 and 6 hours, showing correlation between liberation of CK and histopathological damage. Histopathological alterations were characterized by “delta lesions”, vacuolization, and apparition of empty areas sarcoplasmatic region, also degeneration of muscle fibres appeared. C.officinalis showed protective action versus the miotoxic effect of B. leucurus  venom. The incubation of venom with Callendulaextract suggest the existence of any antivenin fraction in the aquose extract of Callendula. Due to the results we recommend the use of extract of C. officinalis in the form of topical treatment previously to anti-bothropic serum treatment or may be in combination with the proper serum.

Correspondence to: rejane@ufba.br