Carreiro da Costa Roberta S, Prudêncio Luiz, Ferrari Erika Fonseca, Souza Gustavo H M F, de Mello Sueli Moreira, Prianti Júnior Antonio Carlos Guimarães, Ribeiro Wellington, Zamunér Stella Regina, Hyslop Stephen, Cogo José Carlos
Serpentário do Centro de Estudos da Natureza, Instituto de Pesquisa e Desenvolvimento, Universidade do Vale do Paraíba , São José dos Campos, 12244-000, SP, Brazil.
Comp Biochem Physiol C Toxicol Pharmacol. 2008 Jul;148(1):31-8. doi: 10.1016/j.cbpc.2008.03.006. Epub 2008 Mar 14.
Snakes of the opisthoglyphous genus Philodryas are widespread in South America and cause most bites by colubrids in this region. In this study, we examined the neurotoxic and myotoxic effects of venom from Philodryas patagoniensis in biventer cervicis and phrenic nerve-diaphragm preparations and we compared the biochemical activities of venoms from P. patagoniensis and Philodryas olfersii. Philodryas patagoniensis venom (40 microg/mL) had no effect on mouse phrenic nerve-diaphragm preparations but caused time-dependent neuromuscular blockade of chick biventer cervicis preparations. This blockade was not reversed by washing. The highest concentration of venom tested (40 microg/mL) significantly reduced (p<0.05) the contractures to exogenous acetylcholine (55 microM and 110 microM) and K(+) (13.4 mM) after 120 min; lower concentrations of venom had no consistent or significant effect on these responses. Venom caused a concentration- and time-dependent release of creatine kinase (CK) from biventer cervicis preparations. Histological analysis showed contracted muscle fibers at low venom concentrations and myonecrosis at high concentrations. Philodryas venoms had low esterase and phospholipase A(2) but high proteolytic activities compared to the pitviper Bothrops jararaca. SDS-PAGE showed that the Philodryas venoms had similar electrophoretic profiles, with most proteins having a molecular mass of 25-80 kDa. Both of the Philodryas venoms cross-reacted with bothropic antivenom in ELISA, indicating the presence of proteins immunologically related to Bothrops venoms. RP-HPLC of P. patagoniensis venom yielded four major peaks, each of which contained several proteins, as shown by SDS-PAGE. These results indicate that P. patagoniensis venom has neurotoxic and myotoxic components that may contribute to the effects of envenoming by this species.
后沟牙类的菲洛德雷蝰属蛇类在南美洲广泛分布，是该地区游蛇科蛇类咬伤事件的主要致因。在本研究中，我们检测了巴塔哥尼亚菲洛德雷蝰毒液对颈二腹肌和膈神经 - 膈肌标本的神经毒性和肌毒性作用，并比较了巴塔哥尼亚菲洛德雷蝰与奥氏菲洛德雷蝰毒液的生化活性。巴塔哥尼亚菲洛德雷蝰毒液（40微克/毫升）对小鼠膈神经 - 膈肌标本无影响，但对雏鸡颈二腹肌标本引起了时间依赖性的神经肌肉阻滞。这种阻滞不能通过冲洗逆转。所测试的最高毒液浓度（40微克/毫升）在120分钟后显著降低了（p<0.05）对外源性乙酰胆碱（55微摩尔和110微摩尔）和钾离子（13.4毫摩尔）的挛缩反应；较低浓度的毒液对这些反应没有一致或显著的影响。毒液导致颈二腹肌标本中肌酸激酶（CK）呈浓度和时间依赖性释放。组织学分析显示，低毒液浓度下肌肉纤维收缩，高浓度下出现肌坏死。与蝰蛇科的巴西矛头蝮相比，菲洛德雷蝰毒液的酯酶和磷脂酶A2活性较低，但蛋白水解活性较高。SDS - PAGE显示，菲洛德雷蝰毒液具有相似的电泳图谱，大多数蛋白质的分子量为25 - 80 kDa。在ELISA中，两种菲洛德雷蝰毒液均与巴西矛头蝮抗蛇毒血清发生交叉反应，表明存在与巴西矛头蝮毒液免疫相关的蛋白质。巴塔哥尼亚菲洛德雷蝰毒液经反相高效液相色谱分析产生四个主要峰，如SDS - PAGE所示，每个峰都包含几种蛋白质。这些结果表明，巴塔哥尼亚菲洛德雷蝰毒液具有神经毒性和肌毒性成分，可能是该物种致伤作用的原因。
