Key Laboratory of Zoonosis Research by Ministry of Education, College of Veterinary Medicine, Jilin University, Changchun 130062, China.
Department of Parasite, Jilin Academy of Animal Husbandry and Veterinary Medicine, Changchun 130062, China.
Acta Biochim Biophys Sin (Shanghai). 2019 Dec 13;51(12):1286-1292. doi: 10.1093/abbs/gmz128.
Leishmaniasis, caused by the intracellular protozoan parasite Leishmania, remains an important neglected tropical infectious disease. Infection may be lethal if untreated. Currently, the available drugs for the disease are limited by high toxicity and drug resistance. There is an urgent need to develop novel anti-leishmanial strategies. Antimicrobial peptides (AMPs) have been described as the first-line immune defense against pathogenic microbes and are being developed as emerging anti-parasitic therapies. In the present study, we showed the anti-leishmanial activity of the synthetic 4-amino acid peptide lysine, aspartic acid, glutamic acid, and leucine (KDEL), the endoplasmic reticulum retention sequence, against Leishmania tarentolae promastigote and amastigote. Different concentrations of KDEL peptides were incubated with promastigotes, MTT viability assay, and promastigote assay were carried out. Macrophages infected with GFP-transfected L. tarentolae promastigotes were incubated with KDEL peptides, and the anti-amastigote activity of the KDEL peptides was measured by fluorescence microscopy. The damage of L. tarentolae was observed by light microscopy and electron microscopy. The cell apoptosis was analyzed using the Annexin V-FITC/PI apoptosis detection kit and mitochondrial membrane potential assay kit and by flow cytometry. Results showed that L. tarentolae was susceptible to KDEL peptides in a dose-dependent manner, and KDEL peptides disrupted the surface membrane integrity and caused cell apoptosis. In our study, we found for the first time an AMP KDEL from Pseudomonas aeruginosa and proved its significant therapeutic potential as a novel anti-leishmanial drug.
利什曼病是由细胞内原生动物寄生虫利什曼原虫引起的,仍然是一种重要的被忽视的热带传染病。如果不治疗,感染可能是致命的。目前,该疾病可用的药物受到高毒性和耐药性的限制。因此,迫切需要开发新的抗利什曼病策略。抗菌肽(AMPs)被描述为针对致病微生物的第一道免疫防御,并且正在被开发为新兴的抗寄生虫治疗方法。在本研究中,我们展示了合成的 4 氨基酸肽赖氨酸、天冬氨酸、谷氨酸和亮氨酸(KDEL),即内质网保留序列对利什曼原虫前鞭毛体和无鞭毛体的抗利什曼活性。用不同浓度的 KDEL 肽孵育前鞭毛体,进行 MTT 活力测定和前鞭毛体测定。用 GFP 转染的 L. tarentolae 前鞭毛体感染巨噬细胞,用 KDEL 肽孵育,通过荧光显微镜测量 KDEL 肽的抗无鞭毛体活性。用光镜和电镜观察 L. tarentolae 的损伤。用 Annexin V-FITC/PI 凋亡检测试剂盒和线粒体膜电位测定试剂盒以及流式细胞术分析细胞凋亡。结果表明,L. tarentolae 对 KDEL 肽呈剂量依赖性敏感,KDEL 肽破坏表面膜完整性并引起细胞凋亡。在我们的研究中,我们首次从铜绿假单胞菌中发现了一种 AMP KDEL,并证明了其作为新型抗利什曼病药物的显著治疗潜力。