Ferraz Miriéle C, Yoshida Edson H, Tavares Renata V S, Cogo José C, Cintra Adélia C O, Dal Belo Cháriston A, Franco Luiz M, dos Santos Márcio G, Resende Flávia A, Varanda Eliana A, Hyslop Stephen, Puebla Pilar, San Feliciano Arturo, Oshima-Franco Yoko
Post-Graduate Program in Pharmaceutical Sciences, University of Sorocaba (UNISO), Rodovia Raposo Tavares, Km 92.5, 18023-000 Sorocaba, SP, Brazil.
Post-Graduate Program in Technological and Environmental Processes, University of Sorocaba (UNISO), Rodovia Raposo Tavares, Km 92.5, 18023-000 Sorocaba, SP, Brazil.
Molecules. 2014 May 6;19(5):5790-805. doi: 10.3390/molecules19055790.
Snakebite is a neglected disease and serious health problem in Brazil, with most bites being caused by snakes of the genus Bothrops. Although serum therapy is the primary treatment for systemic envenomation, it is generally ineffective in neutralizing the local effects of these venoms. In this work, we examined the ability of 7,8,3'-trihydroxy-4'-methoxyisoflavone (TM), an isoflavone from Dipteryx alata, to neutralize the neurotoxicity (in mouse phrenic nerve-diaphragm preparations) and myotoxicity (assessed by light microscopy) of Bothrops jararacussu snake venom in vitro. The toxicity of TM was assessed using the Salmonella microsome assay (Ames test). Incubation with TM alone (200 μg/mL) did not alter the muscle twitch tension whereas incubation with venom (40 μg/mL) caused irreversible paralysis. Preincubation of TM (200 μg/mL) with venom attenuated the venom-induced neuromuscular blockade by 84% ± 5% (mean ± SEM; n = 4). The neuromuscular blockade caused by bothropstoxin-I (BthTX-I), the major myotoxic PLA2 of this venom, was also attenuated by TM. Histological analysis of diaphragm muscle incubated with TM showed that most fibers were preserved (only 9.2% ± 1.7% were damaged; n = 4) compared to venom alone (50.3% ± 5.4% of fibers damaged; n = 3), and preincubation of TM with venom significantly attenuated the venom-induced damage (only 17% ± 3.4% of fibers damaged; n = 3; p < 0.05 compared to venom alone). TM showed no mutagenicity in the Ames test using Salmonella strains TA98 and TA97a with (+S9) and without (-S9) metabolic activation. These findings indicate that TM is a potentially useful compound for antagonizing the neuromuscular effects (neurotoxicity and myotoxicity) of B. jararacussu venom.
在巴西,蛇咬伤是一种被忽视的疾病和严重的健康问题,大多数咬伤是由具窍蝮蛇属的蛇造成的。尽管血清疗法是全身性中毒的主要治疗方法,但它通常无法有效中和这些毒液的局部作用。在这项研究中,我们检测了来自二翅豆的异黄酮7,8,3'-三羟基-4'-甲氧基异黄酮(TM)在体外中和巴西矛头蝮蛇毒神经毒性(在小鼠膈神经-膈肌标本中)和肌毒性(通过光学显微镜评估)的能力。使用沙门氏菌微粒体试验(艾姆斯试验)评估了TM的毒性。单独用TM(200μg/mL)孵育不会改变肌肉抽搐张力,而用毒液(40μg/mL)孵育会导致不可逆的麻痹。TM(200μg/mL)与毒液预孵育可使毒液诱导的神经肌肉阻滞减弱84%±5%(平均值±标准误;n = 4)。该毒液的主要肌毒性磷脂酶A2——矛头蝮毒素-I(BthTX-I)引起的神经肌肉阻滞也被TM减弱。与单独使用毒液相比(3只小鼠中50.3%±5.4%的纤维受损),用TM孵育的膈肌组织学分析表明,大多数纤维得以保留(4只小鼠中只有9.2%±1.7%受损),并且TM与毒液预孵育可显著减轻毒液诱导的损伤(3只小鼠中只有17%±3.4%的纤维受损;n = 3;与单独使用毒液相比,p < 0.05)。在使用沙门氏菌菌株TA98和TA97a进行的艾姆斯试验中,无论有无(+S9)代谢激活,TM均无致突变性。这些发现表明,TM是一种潜在有用的化合物,可拮抗巴西矛头蝮蛇毒的神经肌肉效应(神经毒性和肌毒性)。