Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Ceará, Fortaleza, CE, Brazil.
Departamento de Fisiologia e Farmacologia, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brazil.
Exp Parasitol. 2020 Aug;215:107930. doi: 10.1016/j.exppara.2020.107930. Epub 2020 May 25.
Trypanosoma cruzi, the etiological agent of Chagas disease, is responsible for the infection of millions of people worldwide and it is a public health problem, without an effective cure. Four fragments with antimicrobial potential from the hemocyanin of Penaeus monodon shrimp were identified using a computer software AMPA. The present study aimed to evaluate the antichagasic effect of these four peptides (Hmc364-382, Hmc666-678, Hmc185-197 and Hmc476-498). The peptides were tested against the epimastigote, trypomastigote and amastigote forms of Trypanosoma cruzi Y strain (benznidazole-resistant strain) and cytotoxicity in mammalian cells was evaluated against LLC-MK2 lineage cells. Two fragments (Hmc364-382, Hmc666-678) showed activity against the epimastigote and trypomastigote forms and their selectivity index (SI) was calculated. The Hmc364-382 peptide was considered the most promising (SI > 50) one and it was used for further studies, using flow cytometry analyses with specific molecular probes and scanning electron microscopy (SEM). Hmc364-382 was able to induce cell death in T. cruzi through necrosis, observed by loss of membrane integrity in flow cytometry analyses and pore formation in SEM. Overall, Hmc364-382 open perspectives to the development of new antichagasic agents.
克氏锥虫,恰加斯病的病原体,导致了全世界数百万人的感染,是一个公共卫生问题,目前尚无有效的治疗方法。本研究使用 AMPA 计算机软件鉴定了来自斑节对虾血蓝蛋白的四个具有抗菌潜力的片段。本研究旨在评估这四个肽(Hmc364-382、Hmc666-678、Hmc185-197 和 Hmc476-498)对克氏锥虫 Y 株(苯并硝唑耐药株)的抗恰加斯病效应。这些肽针对锥虫的前鞭毛体、无鞭毛体和阿米巴样体形式进行了测试,并针对 LLC-MK2 细胞系评估了对哺乳动物细胞的细胞毒性。两个片段(Hmc364-382、Hmc666-678)对前鞭毛体和无鞭毛体形式表现出活性,并计算了其选择性指数(SI)。Hmc364-382 肽被认为是最有前途的(SI>50)一种,并且它被用于进一步的研究,使用特异性分子探针和扫描电子显微镜(SEM)进行流式细胞术分析。Hmc364-382 通过坏死诱导 T. cruzi 细胞死亡,这在流式细胞术分析中观察到细胞膜完整性丧失和 SEM 中孔形成。总的来说,Hmc364-382 为开发新的抗恰加斯病药物开辟了前景。