García-Huertas Paola, Mejía-Jaramillo Ana María, Machado Carlos Renato, Guimarães Anna Cláudia, Triana-Chávez Omar
Grupo Biología y Control de Enfermedades Infecciosas-BCEI, Instituto de Biología, Universidad de Antioquia, Calle 70 52-21, Medellín, Colombia.
Departamento de Bioquímica e Inmunologia, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
R Soc Open Sci. 2017 Sep 20;4(9):170773. doi: 10.1098/rsos.170773. eCollection 2017 Sep.
Nifurtimox (Nfx) and benznidazole (Bz) are the current drugs used for the treatment of Chagas disease. The mechanisms of action and resistance to these drugs in this parasite are poorly known. Prostaglandin F2α synthase or old yellow enzyme (OYE), an NAD(P)H flavin oxidoreductase, has been involved in the activation pathway of other trypanocidal drugs such as Nfx; however, its role in the mechanism of action of Bz is uncertain. In this paper, we performed some experiments of functional genomics in the parasite with the aim to test the role of this gene in the resistance to Bz. For this, we overexpressed this gene in sensitive parasites and evaluated the resistance level to the drug and other chemical compounds such as hydrogen peroxide, methyl methanesulfonate and gamma radiation. Interestingly, parasites overexpressing OYE showed alteration of enzymes associated with oxidative stress protection such as superoxide dismutase A and trypanothione reductase. Furthermore, transfected parasites were more sensitive to drugs, genetic damage and oxidative stress. Additionally, transfected parasites were less infective than wild-type parasites and they showed higher alteration in mitochondrial membrane potential and cell cycle after treatment with Bz. These results supply essential information to help further the understanding of the mechanism of action of Bz in .
硝呋替莫（Nfx）和苯硝唑（Bz）是目前用于治疗恰加斯病的药物。人们对这些药物在该寄生虫中的作用机制和耐药性了解甚少。前列腺素F2α合酶或老黄色酶（OYE）是一种NAD（P）H黄素氧化还原酶，参与了其他杀锥虫药物如Nfx的激活途径；然而，其在Bz作用机制中的作用尚不确定。在本文中，我们在该寄生虫中进行了一些功能基因组学实验，旨在测试该基因在对Bz耐药性中的作用。为此，我们在敏感寄生虫中过表达该基因，并评估对该药物以及其他化合物如过氧化氢、甲磺酸甲酯和γ辐射的耐药水平。有趣的是，过表达OYE的寄生虫显示出与氧化应激保护相关的酶如超氧化物歧化酶A和锥虫硫醇还原酶的改变。此外，转染的寄生虫对药物、基因损伤和氧化应激更敏感。另外，转染的寄生虫比野生型寄生虫的感染力更低，并且在用Bz处理后它们在线粒体膜电位和细胞周期中表现出更高的改变。这些结果提供了重要信息，有助于进一步理解Bz在……中的作用机制。
