Abstract

The use of compounds from natural or synthetic sources and nanotechnology may represent an alternative to develop new drugs for the leishmaniasis treatment. DETC is an inhibitor of the SOD1 enzyme, which leads to increased ROS production, important for the elimination of Leishmania. Thus, our objective was to assess the leishmanicidal in vitro effect of free Diethydithiocarbamate (DETC) and DETC loaded in beeswax-copaiba oil nanoparticles (DETC-Beeswax-CO Nps) on L. amazonensis forms and elucidate the possible mechanisms involved in the parasite death. DETC-Beeswax-CO Nps presented size below 200 nm, spherical morphology, negative zeta potential, and high encapsulation efficiency. Free DETC reduced the viability of promastigotes and increase ROS production, lower the mitochondrial membrane potential, cause phosphatidylserine exposure, and enhance plasma membrane permeability, in addition to promoting morphological changes in the parasite. Free DETC proved toxic in the assessment of toxicity to murine macrophages, however, the encapsulation of this compound was able to reduce these toxic effects on macrophages. DETC-Beeswax-CO Nps exerted anti-amastigote effect by enhancing the production of ROS, superoxide anion, TNF-α, IL-6, and reduced IL-10 in macrophages. Therefore, free DETC induces antipromastigote effect by apoptosis-like; and DETC-Beeswax-CO Nps exerted anti-leishmanial effect due to pro-oxidant and pro-inflammatory response.

摘要

使用来自天然或合成来源的化合物和纳米技术可能代表开发治疗利什曼病的新药物的替代方法。DETC 是 SOD1 酶的抑制剂，可导致 ROS 产生增加，这对消除利什曼原虫很重要。因此，我们的目标是评估游离二乙二硫代氨基甲酸酯 (DETC) 和负载在蜂蜡-copaiba 油纳米粒子 (DETC-Beeswax-CO Nps) 中的 DETC 对L. amazonensis 的体外杀利什曼原虫效果形成并阐明寄生虫死亡的可能机制。DETC-Beeswax-CO Nps 具有小于 200 nm 的尺寸、球形形态、负 zeta 电位和高封装效率。游离的 DETC 降低了前鞭毛体的活力，增加了 ROS 的产生，降低了线粒体膜电位，导致磷脂酰丝氨酸暴露，增强了质膜通透性，此外还促进了寄生虫的形态变化。在评估对小鼠巨噬细胞的毒性时，游离 DETC 被证明是有毒的，然而，这种化合物的封装能够减少这些对巨噬细胞的毒性作用。DETC-Beeswax-CO Nps 通过增强巨噬细胞中 ROS、超氧阴离子、TNF-α、IL-6 和减少 IL-10 的产生来发挥抗无鞭毛体作用。所以，游离 DETC 通过凋亡样诱导抗前鞭毛体作用；由于促氧化和促炎反应，DETC-Beeswax-CO Nps 发挥抗利什曼原虫作用。