RESUMO
O estudo investigou o efeito antiinflamatório do extrato aquoso da Arrabidaea chica (Humb. & Bonpl.) B. Verl., Bignoniaceae, popularmente conhecida como "crajiru", sobre o edema induzido por venenos de serpentes amazônicas dos gêneros Brothrops e Crotalus, em camundongos albinos, por via oral, intraperitoneal e subcutânea. O efeito anti-edematogênico foi avaliado pela medição do diâmetro dos coxins das patas posteriores, sendo medidos as 1, 3, 6, 12 e 24 horas, para B. atrox e 1, 3 e 6 horas para Crotalus durissus ruruima, e também avaliado por histopatologia. O estudo mostrou que o efeito inibitório do extrato aquoso para o gênero Bothrops, pelas vias subcutânea e intraperitoneal (12 horas) foi de 55,87 por cento e 65,70 por cento, respectivamente. Para o gênero Crotalus o efeito inibitório do extrato pela via subcutânea após 3 horas foi de 33,55 por cento e após 6 horas de 79,81 por cento. Pela via intraperitoneal após 3 horas foi de 48,02 por cento e após 6 horas de 92,52 por cento. Na análise histopatológica, o infiltrado de granulócitos e a miocitólise foram os efeitos inflamatórios inibidos mais significativamente. Os resultados sugerem a presença no extrato aquoso de A. chica de substâncias com atividade inibitória sobre os efeitos inflamatórios dos venenos das serpentes Bothrops atrox e Crotalus durissus ruruima.
The study investigated the anti-inflammatory effects of the aqueous extract (EAq), the Arrabidaea chica (Humb. & Bonpl.) B. Verl., Bignoniaceae, known as the "crajiru", tested on the paw oedema induced in inflammatory process by venoms Amazon snakes from the Bothrops and Crotalus species, on albino mice. This process was done by an oral, intraperitoneal and subcutaneous way. The cushions thickness was measured at time breaks of 1, 3, 6, 12 and 24 hours, Bothrops genus and 1, 3 and 6 hours, Crotalus genus, and a histopathologic analysis was made. The Bothrops genus, was used as an inhibitory effect for the subcutaneous and intraperitoneal way (12 hours), and it had an inhibition of 55.87 percent and 65.70 percent, respectively. For the Crotalus genus, the inhibitory effect for the subcutaneous way after 3 hours was the 33.55 percent and 79.81 percent, after 6 hours. The intraperitoneal way affect after 3 hours had an inhibition of 48.02 percent and 92.52 percent after 6 hours. For the histopathological analysis, the leucocytes infiltration and the miocitolisis were the most expressive parameters and thus inhibited. The results suggest the presence of active principles with anti-inflammatory effects.
RESUMO
A comparative study of venoms from juvenile, sub-adult and adult wild Bothrops atrox specimens captured in Manaus region (Brazil) was performed. All venoms tested had acidic pH (5.5) and the human plasma coagulant activity was higher in venoms from juvenile and sub-adult specimens than in adults. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the most intense bands in adult venoms corresponded to polypeptides of 23 and 50kDa. The 23kDa protein was not detected in juvenile venoms. The 23 and 50kDa proteins were purified by two steps of reversed phase-HPLC followed by size exclusion HPLC. Partial amino acid sequence of the 23kDa protein showed homology to metalloproteinases from other snake venoms. Electrospray ionization mass spectrometric analysis (ESI-MS) showed that the 23kDa band contained at least three isoforms of 23030, 23300 and 23645Da. The 50kDa polypeptide was N-terminally blocked for Edman degradation and presented molecular masses ranging from 46.8 to 49.4kDa by ESI-MS. Both proteins were detected by anti-mutalysin II antibodies in immunoblotting assay indicating that they belong to the metalloproteinase family. Immunoblotting analysis also showed that the 23kDa band increased in intensity from juvenile to adult specimens.SDS-PAGE analysis of juvenile and adult venoms following autoproteolysis in pH 7.4 suggested that endogenous venom metalloproteinases can digest the 50kDa metalloproteinase, originating a new protein band of 27kDa. It was also demonstrated in juvenile venoms that the 23kDa band was not the result of proteolytic processing of the 50kDa metalloproteinase.