重组α毒素抗利什曼原虫作用的评估

Evaluation of the Anti-Leishmanial Effect of Recombinant α-Toxin.

作者信息

Namdar Fatemeh, Khanahmad Hossein, Ghayour Zahra, Mirzaei Farzaneh, Namdar Azam, Aghaei Maryam, Izadi Shahrokh, Khamesipour Faham, Hejazi Seyed Hossein

机构信息

Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Infect Drug Resist. 2020 Jul 15;13:2355-2364. doi: 10.2147/IDR.S257561. eCollection 2020.

DOI:10.2147/IDR.S257561

PMID:32765010

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7369417/

Abstract

BACKGROUND

Leishmaniasis is an infectious disease common in tropical and subtropical regions caused by the genus , which is transmitted by the bite of female sandflies. In this study, we evaluate the anti-leishmanial effect of recombinant α-toxin protein alone and the combination with glucantime through in vitro and in vivo.

MATERIALS AND METHODS

Production, expression, and purification of recombinant α-toxin were evaluated by SDS-PAGE and Western blotting techniques. The antileishmanial activities of the purified α-toxin plus and without glucantime were examined in vitro and in vivo.

RESULTS

The results indicated successful expression of α-toxin as a 48 kDa band on SDS-PAGE and Western blot methods. Also, evaluation of α-toxin IC showed the strong fatal effect of it, and glucantime on medium proliferated promastigotes at lower concentrations compared with glucantime or α-toxin alone. Moreover, in vivo surveys showed that at the end of treatment courses, the mean of lesion size diminished in glucantime plus α-toxin treated mice versus negative control groups (p < 0.001). Also, there was a significant difference in the parasite burden of the spleen and liver of the control versus the test groups (p < 0.001).

CONCLUSION

The results showed recombinant α-toxin has synergistic effects with glucantime in destroying parasites.

摘要

背景

利什曼病是一种在热带和亚热带地区常见的传染病，由利什曼原虫属引起，通过雌性白蛉叮咬传播。在本研究中，我们通过体外和体内实验评估重组α-毒素蛋白单独以及与葡糖胺联合使用的抗利什曼原虫效果。

材料与方法

通过SDS-PAGE和蛋白质印迹技术评估重组α-毒素的产生、表达和纯化。在体外和体内检测纯化的α-毒素加或不加葡糖胺的抗利什曼原虫活性。

结果

结果表明，在SDS-PAGE和蛋白质印迹方法中，α-毒素成功表达为一条48 kDa的条带。此外，α-毒素IC评估显示其具有强大的致死作用，与单独使用葡糖胺或α-毒素相比，葡糖胺与α-毒素联合使用在较低浓度下对培养基中增殖的前鞭毛体有更强的作用。此外，体内研究表明，在治疗疗程结束时，与阴性对照组相比，葡糖胺加α-毒素治疗的小鼠病变大小平均值减小（p < 0.001）。而且，对照组与试验组脾脏和肝脏的寄生虫负荷存在显著差异（p < 0.001）。