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抑制红口蝮蛇毒诱导的局部效应:评估巴西多价抗蝮蛇毒血清和棉叶麻风树水提物的有效性。

Inhibition of local effects induced by Bothrops erythromelas snake venom: Assessment of the effectiveness of Brazilian polyvalent bothropic antivenom and aqueous leaf extract of Jatropha gossypiifolia.

作者信息

Félix-Silva Juliana, Gomes Jacyra A S, Xavier-Santos Jacinthia B, Passos Júlia G R, Silva-Junior Arnóbio A, Tambourgi Denise V, Fernandes-Pedrosa Matheus F

机构信息

Laboratório de Tecnologia & Biotecnologia, Faculdade de Farmácia, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.

Laboratório de Imunoquímica, Instituto Butantan, São Paulo, SP, Brazil.

出版信息

Toxicon. 2017 Jan;125:74-83. doi: 10.1016/j.toxicon.2016.11.260. Epub 2016 Nov 25.

Abstract

Bothrops erythromelas is a snake of medical importance responsible for most of the venomous incidents in Northeastern Brazil. However, this species is not included in the pool of venoms that are used in the Brazilian polyvalent bothropic antivenom (BAv) production. Furthermore, it is well known that antivenom therapy has limited efficacy against venom-induced local effects, making the search for complementary alternatives to treat snakebites an important task. Jatropha gossypiifolia is a medicinal plant widely indicated in folk medicine as an antidote for snakebites, whose effectiveness against Bothrops jararaca venom (BjV) has been previously demonstrated in mice. In this context, this study assessed the effectiveness of the aqueous extract (AE) of this plant and of the BAv against local effects induced by B. erythromelas venom (BeV). Inhibition of BeV-induced edematogenic and hemorrhagic local effects was assayed in mice in pre-treatment (treatment prior to BeV injection) and post-treatment (treatment post-envenomation) protocols. Inhibition of proteolytic, phospholipase A (PLA) and hyaluronidase enzymatic activities of BeV were evaluated in vitro. BAv cross-reactivity and estimation of antibody titers against BeV and BjV were assessed by Ouchterlony double diffusion test. The results show that in pre-treatment protocol AE and BAv presented very similar effects (about 70% of inhibition for edematogenic and 40% for hemorrhagic activities). However, BAv poorly inhibited edema and hemorrhage in post-envenomation protocol, whilst, in contrast, AE was significantly active even when used after BeV injection. AE was able to inhibit all the tested enzymatic activities of BeV, while BAv was active only against hyaluronidase activity, which could justify the low effectiveness of BAv against BeV-induced local effects in vivo. Ouchterlony's test showed positive cross-reactivity against BeV, but the antibody titers were slightly higher against BjV. Together, these data indicate that despite the presence of immunological cross-reactivity, Brazilian polyvalent bothropic antivenom presented low inhibitory potential against biological and enzymatic effects of BeV, illustrating the need for new strategies in the production of antivenom with broad neutralizing potential in the treatment of Bothrops spp. envenomation throughout the country. Together, the results highlight the antiophidic potential of J. gossypiifolia, suggesting that it can be considered a potential adjuvant in the treatment of bothropic envenomation local effects.

摘要

红口蝮是一种具有医学重要性的蛇类,在巴西东北部造成了大多数毒蛇咬伤事件。然而,该物种并不在巴西多价抗红口蝮蛇毒血清(BAv)生产所使用的毒液库中。此外,众所周知,抗蛇毒血清疗法对毒液引起的局部效应疗效有限,因此寻找治疗蛇咬伤的补充替代方法成为一项重要任务。棉叶麻风树是一种药用植物,在民间医学中被广泛用作蛇咬伤的解毒剂,其对巴西矛头蝮蛇毒(BjV)的有效性先前已在小鼠身上得到证实。在此背景下,本研究评估了该植物的水提取物(AE)和BAv对红口蝮蛇毒(BeV)引起的局部效应的有效性。在预处理(BeV注射前治疗)和后处理(中毒后治疗)方案中,在小鼠身上测定了对BeV诱导的致水肿和出血性局部效应的抑制作用。在体外评估了对BeV的蛋白水解、磷脂酶A(PLA)和透明质酸酶酶活性的抑制作用。通过双向琼脂扩散试验评估了BAv的交叉反应性以及针对BeV和BjV的抗体滴度。结果表明,在预处理方案中,AE和BAv呈现出非常相似的效果(致水肿活性抑制约70%,出血活性抑制约40%)。然而,在中毒后方案中,BAv对水肿和出血的抑制作用较差,而相比之下,即使在BeV注射后使用,AE仍具有显著活性。AE能够抑制BeV的所有测试酶活性,而BAv仅对透明质酸酶活性有活性,这可以解释BAv在体内对BeV诱导的局部效应有效性较低的原因。双向琼脂扩散试验显示对BeV有阳性交叉反应,但对BjV的抗体滴度略高。总之,这些数据表明,尽管存在免疫交叉反应性,但巴西多价抗红口蝮蛇毒血清对BeV的生物学和酶学效应的抑制潜力较低,这说明在全国范围内生产具有广泛中和潜力的抗蛇毒血清以治疗矛头蝮属蛇咬伤时需要新的策略。总之,结果突出了棉叶麻风树的抗蛇毒潜力,表明它可被视为治疗红口蝮蛇咬伤局部效应的潜在佐剂。

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