Identification of antinociceptive fraction of snake venom from Crotalus durissus collilineatus crotamine-negative and its acute toxicity evaluation

Abstract

The crude venom of the snake Crotalus durissus collilineatus (CDC) promotes neurological signs and symptoms in accidents involving humans and animals and the victims reports analgesia at the bite site, without tissue destruction. Studies shows that CDC has analgesic activity, among others. The crude venom is considered unsuitable for therapeutic purposes, with encouragement to the fractionation and purification of the same. Thus, the aim with CDC venom is: to perform fractionation by preparative HPLC; to test the antinociceptive activity of fractions and acute toxicity of active fractions. The CDC was fractionated on preparative HPLC-PDA (Oliveira et al., 2015) and the fractions were tested for their antinociceptive activity for writhing test by acetic acid (0.6%) in mice. For one of the fractions, which showed high analgesic effect both p.o. and i.p. routes, it evaluated the acute toxicity by the up and down method (OECD, 2001). In the fractionation by HPLC-PDA, CDC yielded 10 peaks (P1P10). SDS-PAGE showed that there was a good separation of components of the venom. All peaks were evaluated for their ability to reduce writhing, and the only one that apparently showed antinociceptive effect was Fr5 fraction (40 μg/kg). The Fr5 was able to reduce by 47% the number of contortions (i.p.) and 87% (p.o.), compared to control. The Fr5 fraction showed no morbidity and no mortality in the acute toxicity test (dose of 1000 μg/kg, p.o.); so it was not possible to estimate the LD50. According to the results, it can be stated that the venom and Fr5 of Crotalus durissus collilineatus snake of crotamine-negative type, may exhibit antinociceptive activity by suppressing nociception induced by acetic acid, suggesting it is related to effects on peripheral sites spinal and presents low acute toxicity values in experimental animals.

Antinociceptive and Anti-Inflammatory Effects of Recombinant Crotamine in Mouse Models of Pain

“…Control animals received only saline (100 µL). After 20 min [14], 100 µL of a 0.6% (v/v) acetic-acid solution was intraperitoneally administered, and mice were immediately returned to the transparent acrylic observation cage (Supplementary Figure S1). The number of writhes was then measured for 20 min [15].…”

A Review of Rattlesnake Venoms

“…Furthermore, previous studies have shown that members of this toxin family, specifically crotamine, are expressed differently amongst individuals and populations of Crotalus durissus [ 33 , 271 , 272 ]. Although the family has been discovered for over 50 years, many of its activities have been discovered in recent decades, with several more potential therapeutic applications [ 273 ].…”

A Meta-Analysis of the Protein Components in Rattlesnake Venom

“…41 studies reported the relative abundance of protein constituents. [40] C. cerastes 3FTx, 5 NT, BPP, CRiSP, CTL, Dis, ficolin, Hya, Kun, LAAO, MYO, NGF, PDE, PLA 2 , SVMP P-II/III, SVSP, VEGF, vespryn, WAP [98,[100][101][102] C. durissus 3FTx, achase, aminopeptidase, angiogenin, BPP, carboxypeptidase, CNP, CRiSP, CTL, CysProt inhibitor, CysProt, dipeptidyl peptidase, Dis, FGF, fraction 5, Hya, Kazal, Kun, LAAO, lipase, MYO, NGF, PDGF, PLA 2 (non-CRTX, CRTX), PLB, PLD, Serpin-like, SVMP inhibitor, SVMP P-III, SVSP, VEGF, vespryn, WAP [45,64,[103][104][105][106][107][108][109][110][111][112][113][114][115][116][117][118] C. enyo SVSP, SVMP P-I/III, PLA 2 [40] C. horridus 5 -NT, BPP, CNP, CRiSP, Dis, EGF-like, GC, Hya, Kun, LAAO, MYO, neurotrophic factor, NGF, PDE, PLA 2 , SVMP P-I/III, SVSP, VEGF, vespryn [29,119,120] C. lepidus 5 NT, CRiSP, CTL, Dis, LAAO, PDE, PLA 2 , SVMP-P-I/III, SVSP (TLE, kallikrein) [49,[121][122][123] C. mitchelli LAAO, SVSP, PLA 2 (CRTX/MTX) [40,124,125] C. molossus Dis, LAAO, MYO, PLA 2 , SVMP P-I/III, SVSP (TLE) [58,98,[126][127][128][129][130]…”