Ghobakhloo Nafiseh, Motazedian Mohammad Hossein, Fardaei Majid
Dept. of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Basic Sciences in Infectious Diseases, Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Iran J Parasitol. 2016 Apr-Jun;11(2):168-176.
Treatment of Cutaneous Leishmaniasis (CL) is being faced with serious difficulties in Fars Province, due to emerging of resistance against meglumine antimonite (Glucantime®). In this context, determining some biomarkers for drug sensitivity monitoring seems to be highly essential. Different studies have been carried out to decipher the genes might be involved in antimony resistant phenotype in spp. Here, we selected three genes: AQP (as drug transporter), TDR-1-1(as drug activator), and γ-GCS (inducing reduction environment) for comparative expression analysis on clinical resistant and sensitive isolates of .
The clinical isolates of were collected from CL patients referred to Valfajr Health Center, Shiraz from Oct 2011 to Feb 2012. The susceptibility test was performed to confirm drug sensitivity of strains in vitro as well. Then, the gene expression analysis was performed by quantitative real-time PCR using SYBR® Green.
By comparison of expression level between strains, up regulation of γ-GCS gene and down regulation of AQP gene were observed in resistant strains compared to the sensitive isolates; however, down regulation of AQP was not statistically specific. Analysis of TDR-1-1 gene unexpectedly showed a high level of expression in the non-responsive cases.
The γ-GCS, at least, can be considered as a suitable molecular marker for screening antimony sensitivity in clinical isolates, although AQP and TDR-1-1gene seem not to be reliable resistant markers.
由于对葡甲胺锑酸盐（葡糖胺锑钠®）出现耐药性，法尔斯省皮肤利什曼病（CL）的治疗面临严重困难。在此背景下，确定一些用于药物敏感性监测的生物标志物显得至关重要。已经开展了不同的研究来破译可能参与利什曼原虫锑耐药表型的基因。在这里，我们选择了三个基因：水通道蛋白（AQP，作为药物转运体）、TDR-1-1（作为药物激活剂）和γ-谷氨酰半胱氨酸合成酶（γ-GCS，诱导还原环境），用于对利什曼原虫临床耐药和敏感分离株进行比较表达分析。
2011年10月至2012年2月期间，从设拉子瓦尔法吉尔健康中心转诊的CL患者中收集利什曼原虫临床分离株。还进行了药敏试验以确认菌株在体外的药物敏感性。然后，使用SYBR® Green通过定量实时PCR进行基因表达分析。
通过比较菌株之间的表达水平，与敏感分离株相比，耐药菌株中观察到γ-GCS基因上调和AQP基因下调；然而，AQP的下调在统计学上并不具有特异性。TDR-1-1基因分析意外地显示在无反应病例中表达水平较高。
至少γ-GCS可被视为临床分离株中筛选锑敏感性的合适分子标志物，尽管AQP和TDR-1-1基因似乎不是可靠的耐药标志物。
