Amiri-Dashatan Nasrin, Koushki Mehdi, Rezaei-Tavirani Mostafa, Ahmadi Nayebalia
Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Clinical Biochemistry, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Rep Biochem Mol Biol. 2020 Oct;9(3):324-330. doi: 10.29252/rbmb.9.3.324.
and are the etiological agents of cutaneous leishmaniosis. Leishmania species cause a board spectrum of phenotypes. A small number of genes are differentially expressed between them that have likely an important role in the disease phenotype. Procyclic and metacyclic are two morphological promastigote forms of that express different genes. The glutathione peroxidase is an important antioxidant enzyme that essential in parasite protection against oxidative stress and parasite survival. This study aimed to compare glutathione peroxidase (TDPX) gene expression in procyclic and metacyclic and also interspecies in Iranian isolates of and .
The samples were cultured in Novy-Nicolle-Mc Neal medium to obtain the promastigotes and identified using PCR-RFLP technique. They were then grown in RPMI1640 media for mass cultivation. The expression level of TDPX gene was compared by Real-time PCR.
By comparison of expression level, up-regulation of TDPX gene was observed (5.37 and 2.29 folds) in and metacyclic compared to their procyclic, respectively. Moreover, there was no significant difference between procyclic forms of isolates, while 3.05 folds up-regulation in metacyclic was detected in L. major compared L. tropica.
Our data provide a foundation for identifying infectivity and high survival related factors in the spp. In addition, the results improve our understanding of the molecular basis of metacyclogenesis and development of new potential targets to control or treatment and also, to the identification of species-specific factors contributing to virulence and pathogenicity in the host cells.
硕大利什曼原虫和热带利什曼原虫是皮肤利什曼病的病原体。利什曼原虫物种会引发一系列广泛的表型。它们之间有少数基因差异表达，这些基因可能在疾病表型中发挥重要作用。前鞭毛体和后鞭毛体是利什曼原虫的两种形态前鞭毛体形式，表达不同的基因。谷胱甘肽过氧化物酶是一种重要的抗氧化酶，对寄生虫抵御氧化应激和生存至关重要。本研究旨在比较伊朗分离的硕大利什曼原虫和热带利什曼原虫前鞭毛体和后鞭毛体中谷胱甘肽过氧化物酶（TDPX）基因的表达情况，以及种间差异。
将样本在诺维 - 尼科尔 - 麦克尼尔培养基中培养以获得前鞭毛体，并使用PCR - RFLP技术进行鉴定。然后将它们在RPMI1640培养基中培养以进行大规模培养。通过实时PCR比较TDPX基因的表达水平。
通过比较表达水平，发现硕大利什曼原虫和热带利什曼原虫的后鞭毛体中TDPX基因分别比其前鞭毛体上调（5.37倍和2.29倍）。此外，分离株的前鞭毛体形式之间没有显著差异，而与热带利什曼原虫相比，硕大利什曼原虫的后鞭毛体中检测到上调3.05倍。
我们的数据为鉴定利什曼原虫属物种中的感染性和高生存相关因素提供了基础。此外，结果增进了我们对后鞭毛体形成的分子基础以及控制或治疗新潜在靶点的理解，同时也有助于鉴定宿主细胞中毒力和致病性的物种特异性因素。
