ANTIHEMORRHAGIC ACTIVITY FROM Schizolobium parahyba FRACTIONS CAUSED BY Bothrops alternatus SNAKE VENOM

VALE L. H. F. (1), MENDES M. M (1), RODRIGUES V. M (1), HAMAGUCHI A. (1), PEREIRA P. S. (2), FRANÇA S. C. (2), HOMSI-BRANDEBURGO M. I. (1).

(1) Laboratório de Toxinas Animais e Inibidores, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, Brasil; (2) Departamento de Biotecnologia de Plantas Medicinais, Universidade de Ribeirão Preto, Ribeirão Preto, Brasil.

Plant extracts constitute an extremely rich source of biologically active compounds, and a number of extracts have been shown to possess antivenom activity. This study reports the ability of the fractions of Schizolobium parahyba to inhibit hemorrhagic activity of Bothrops alternatus snake venom. The crude aqueous extract (AE) was prepared from the leaves by mixing in destilled water, centrifuged and the supernatant was lyophilized and stored at -20oC. AE (10g) was re-solubilized in methanol, centrifuged and the supernatant was evaporated (AEM). AEM (7g) was submitted to fractionation on Sephadex LH-20 and eluted with methanol. Seven fractions were collected. Fractions four and five were subimitted to re-chomatography and eleven subfractions were obtained (SPM4.1 to SPM4.5 and SPM5C.1 to SPM5C.6 respectively). These all fractions were submitted to analytic HPLC to analyze the purification degree and possible groups in which this substances belong. Four fractions showed antihemorrhagic activities.  Rutin, an antihemorrhage flavonol, was tested too for comparison to SPM4.3 because this fraction has high amount of Rutin. SPM4.3 was tested too in pre-treatment assays (30 min) because Rutin has vessel constriction action and no inhibitory action on hemorrhagical metaloproteinase enzymes. The inhibition of hemorrhage effect was high in the pre-treatment. We concluded that S. parahyba aqueous extract possess active compounds with bothropic hemorrhagic  action of easy purification in resin Sephadex LH-20 and this compounds resemble to belong to a group of phenolic substances as flavonols and catechins.

KEY WORDS: Schizolobium parahyba, Bothrops alternatus, fractionation, HPLC, antivenom, natural compounds.

FINANCIAL SUPPORT: FAPESP, CAPES, CNPq, FAPEMIG, UFU.

CORRESPONDENCE TO: Avenida Pará, 1720, Bairro Umuarama, Uberlândia-MG, cep: 38405-320. E-mail: lhvale@hotmail.com