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博氏矛头蝮蛇 venom 中的 Lys49-磷脂酶 A2 同源物 BnSP-6 对委内瑞拉旋毛线虫的驱虫作用

The Anthelmintic Effect on Strongyloides venezuelensis Induced by BnSP- 6, a Lys49-phospholipase A2 Homologue from Bothrops pauloensis Venom.

机构信息

Laboratory of Nanobiotechnology, Institute of Biotechnology, Federal University of Uberlandia, Av. Amazonas s/n, Campus Umuarama BL-2E, SL-248, Uberlândia, Minas Gerais, 38400-902, Brazil.

Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlandia, Av. Amazonas s/n, Campus Umuarama BL-2E, SL-227, Uberlândia, Minas Gerais, 38400-902, Brazil.

出版信息

Curr Top Med Chem. 2019;19(22):2032-2040. doi: 10.2174/1568026619666190723152520.

Abstract

BACKGROUND

Phospholipases A2 (PLA2) from snake venoms have a broad potential as pharmacological tools on medicine. In this context, strongyloidiasis is a neglected parasitic disease caused by helminths of the genus Strongyloides. Currently, ivermectin is the drug of choice for treatment, however, besides its notable toxicity, therapeutic failures and cases of drug resistance have been reported. BnSP-6, from Bothorps pauloensis snake venom, is a PLA2 with depth biochemical characterization, reporting effects against tumor cells and bacteria.

OBJECTIVE

The aim of this study is to demonstrate for the first time the action of the PLA2 on Strongyloides venezuelensis.

METHODS

After 72 hours of treatment with BnSP-6 mortality of the infective larvae was assessed by motility assay. Cell and parasite viability was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Furthermore, autophagic vacuoles were labeled with Monodansylcadaverine (MDC) and nuclei of apoptotic cells were labeled with Propidium Iodide (PI). Tissue degeneration of the parasite was highlighted by Transmission Electron Microscopy (TEM).

RESULTS

The mortality index demonstrated that BnSP-6 abolishes the motility of the parasite. In addition, the MTT assay attested the cytotoxicity of BnSP-6 at lower concentrations when compared with ivermectin, while autophagic and apoptosis processes were confirmed. Moreover, the anthelmintic effect was demonstrated by tissue degeneration observed by TEM. Furthermore, we report that BnSP-6 showed low cytotoxicity on human intestinal cells (Caco-2).

CONCLUSION

Altogether, our results shed light on the potential of BNSP-6 as an anthelmintic agent, which can lead to further investigations as a tool for pharmaceutical discoveries.

摘要

背景

蛇毒液中的磷脂酶 A2(PLA2)在医学上具有广泛的作为药理学工具的潜力。在这种情况下,钩虫病是一种被忽视的寄生虫病,由旋毛虫属的蠕虫引起。目前,伊维菌素是首选的治疗药物,但其毒性明显,治疗失败和耐药病例时有报道。BnSP-6 来自 Bothorps pauloensis 蛇毒,是一种 PLA2,具有深入的生化特性,对肿瘤细胞和细菌有作用。

目的

本研究旨在首次证明 PLA2 对委内瑞拉旋毛虫的作用。

方法

用 BnSP-6 处理 72 小时后,通过运动试验评估感染幼虫的死亡率。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物(MTT)测定法评估细胞和寄生虫的活力。此外,用单丹磺酰尸胺(MDC)标记自噬空泡,用碘化丙啶(PI)标记凋亡细胞的核。用透射电子显微镜(TEM)突出寄生虫的组织退化。

结果

死亡率指数表明 BnSP-6 可消除寄生虫的运动能力。此外,MTT 试验表明,与伊维菌素相比,BnSP-6 在较低浓度时具有细胞毒性,同时证实了自噬和凋亡过程。此外,通过 TEM 观察到的组织退化证明了驱虫作用。此外,我们报告 BnSP-6 对人肠细胞(Caco-2)显示出低细胞毒性。

结论

总之,我们的结果表明 BNSP-6 作为驱虫剂具有潜力,可以进一步研究作为药物发现的工具。

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