Crotalus Neutralizing Factor (CNF) inhibits the toxic effects of Crotoxin at mouse neuromuscular preparations.

Crotalus Neutralizing Factor (CNF) was the first phospholipase A inhibitor isolated from the plasma of the South American rattlesnake (Crotalus durissus terrificus). Previous biochemical and biophysical studies demonstrate an interaction of CNF with Crotoxin (CTX), the main toxic component in the venom of these snakes. CTX promotes the blockade of neuromuscular transmission by a sum of neurotoxic and myotoxic activities. However, the ability of CNF to inhibit these activities has not been shown until the present study. We performed a myographic study to compare the neuromuscular effects of CTX and the mixture CTX plus CNF in mice phrenic nerve-diaphragm muscle preparations. CTX (5 μg/mL) alone, or pre-incubated with CNF (5, 20 or 50 μg/mL) for 15 min was added to the preparations and maintained throughout the experimentation period. Myotoxicity was assessed by light microscopic analysis of diaphragm muscle after myographic study. CTX (5 μg/mL) blocked both indirectly and directly evoked twitches in neuromuscular preparations. In addition, CTX induced histological alterations in diaphragm muscle. Pre-incubation with CNF (50 μg/mL) abolished both the muscle-paralyzing and muscle-damaging activities of CTX. Therefore, the present study confirms, through functional studies, the antiophidic potential of CNF.