Myonecrosis induced in rat by a myotoxin isolated from the venom of Bothrops neuwiedifrom Alfenas, MG

Myonecrosis induced in rat by a myotoxin isolated from the venom of Bothrops neuwiedifrom Alfenas, MG

Leite, V.L.; Andrade, M.F.; Lúcia, M.; Gouvêa, C.M. C. P.^*

Laboratório de Bioquímica e Biologia Molecular, Universidade de Alfenas, Alfenas, MG

ABSTRACT

Bothrops venoms are composed by several protein fractions. Among these fractions there are myotoxins which induce an important muscle lesion. The purification of this component involves some steps, although providing a pure material, is time consuming. In the present study, we describe a quick method for myotoxin fraction isolation from the venom of Bolhrops nsuwiedi using one-step high performance liquid chromatography (HPLC). The complete procedure took 30 min. The pur6+y of the two myotoxic fractions isolated was assessed by SDS-PAGE. Upon i.m. injection into rat tibialis anterior, both toxins induced early morphological changes, indicating that the plasma membrane was the first cellular structure affected. Afterwards, the lesion was typically myonecrotic and inflammatory infiltrate was present.

Key words:Bothrops neuwiedi, HPLC, myonecrosis, myotoxin

RESUMO

Os venenos de Bolhrops são compostos por várias frações protéicas. Dentre elas, as miotoxinas induzem lesões musculares importantes. A purificação deste componente envolve várias etapas e, embora forneça frações puras, é muito trabalhosa. Neste trabalho foi descrito um método rápido para o isolamento da fração miotóxica do veneno de Bothrops neuwiedi (jararaca pintada), utilizando-se uma etapa única de purificação por Cromatografia Liquida de Alta Eficiência (CLAE). O procedimento completo demorou apenas 30 min. Foram isoladas duas frações com atividade miotóxica, cuja pureza foi verificada por eletroforese em gel de poliacrilamida-SDS. A injeção intramuscular no músculo tibial anterior de ratos induziu alterações morfológicas precoces, indicando que a membrana plasmática foi a primeira estrutura celular afetada pela fração miotóxica. A lesão muscular obtida foi tipicamente mionecrótica e foi observado infiltrado inflamatório.

Unitermos:Bothrops neuwiedi, CLAE, mionecrose, miotoxina

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ACKNOWLEGEMENTS

We thank Prof. Dr. José Carlos Tavares Carvalho from his helpful assistance with HPLC.

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Publication Dates

• Publication in this collection
  05 Oct 2009
• Date of issue
  2001