Tiuman Tatiana Shioji, Ueda-Nakamura Tânia, Alonso Antonio, Nakamura Celso Vataru
Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Estadual de Maringá, Av, Colombo 5790, 87020-900 Maringá, Paraná, Brazil.
BMC Microbiol. 2014 Jun 10;14:152. doi: 10.1186/1471-2180-14-152.
Leishmania amazonensis infection results in diverse clinical manifestations: cutaneous, mucocutaneous or visceral leishmaniasis. The arsenal of drugs available for treating Leishmania infections is limited. Therefore, new, effective, and less toxic leishmaniasis treatments are still needed. We verified cell death in amastigote forms of Leishmania amazonensis induced by the sesquiterpene lactone parthenolide.
The tested compound was able to concentration-dependently affect axenic and intracellular amastigotes, with IC50 values of 1.3 μM and 2.9 μM, respectively after 72 h incubation. No genotoxic effects were observed in a micronucleus test in mice. Parthenolide induced morphological and ultrastructural changes in axenic amastigotes, including a loss of membrane integrity, swelling of the mitochondrion, cytoplasmic vacuoles, and intense exocytic activity in the region of the flagellar pocket. These results led us to investigate the occurrence of autophagic vacuoles with monodansylcadaverine and the integrity of the plasma membrane and mitochondrial membrane potential using flow cytometry. In all of the tests, parthenolide had positive results.
Our results indicate that the antileishmanial action of parthenolide is associated with autophagic vacuole appearance, a reduction of fluidity, a loss of membrane integrity, and mitochondrial dysfunction. Considering the limited repertoire of existing antileishmanial compounds, the products derived from medicinal plants has been one the greatest advances to help develop new chemotherapeutic approaches.
亚马逊利什曼原虫感染会导致多种临床表现:皮肤利什曼病、黏膜皮肤利什曼病或内脏利什曼病。可用于治疗利什曼原虫感染的药物种类有限。因此,仍然需要新的、有效且毒性较小的利什曼病治疗方法。我们验证了倍半萜内酯小白菊内酯诱导的亚马逊利什曼原虫无鞭毛体形式的细胞死亡。
经测试的化合物能够浓度依赖性地影响体外培养和细胞内的无鞭毛体,孵育72小时后,其IC50值分别为1.3 μM和2.9 μM。在小鼠微核试验中未观察到遗传毒性作用。小白菊内酯诱导体外培养的无鞭毛体发生形态和超微结构变化,包括膜完整性丧失、线粒体肿胀、细胞质空泡形成以及鞭毛袋区域强烈的胞吐活性。这些结果促使我们使用单丹磺酰尸胺研究自噬泡的出现情况,并通过流式细胞术研究质膜完整性和线粒体膜电位。在所有测试中,小白菊内酯均得到阳性结果。
我们的结果表明,小白菊内酯的抗利什曼原虫作用与自噬泡的出现、流动性降低、膜完整性丧失以及线粒体功能障碍有关。考虑到现有的抗利什曼原虫化合物种类有限,药用植物衍生产品一直是帮助开发新化疗方法的最大进展之一。