Koutsoni Olga S, Karampetsou Kalliopi, Dotsika Eleni
Laboratory of Cellular Immunology & National Reference Laboratory for Leishmaniasis, Department of Microbiology, Hellenic Pasteur Institute, 127 Vass. Sofias av., 11521 Athens, Greece.
Division of Pharmacognosy and Natural Products Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771 Athens, Greece.
Bio Protoc. 2019 Nov 5;9(21):e3410. doi: 10.21769/BioProtoc.3410.
Neglected tropical diseases gain the scientific interest of numerous research programs in an attempt to achieve their effective control or elimination. In this attempt, more cutting-edge public health policies and research are needed for the discovery of new, safer and effective drugs originated from natural products. Here, we describe protocols for the screening of a natural product-derived compound required for the determination of its antileishmanial potency. For this purpose, the Total Phenolic Fraction (TPF) derived from extra virgin olive oil is evaluated through the cell culture method against extracellular promastigote and intracellular amastigote spp. forms. The aim of this article is to describe a step-by-step procedure that can be easily applied to accurately estimate the 50% inhibitory concentration (IC), the 50% cytotoxic concentration (CC) and the selectivity index (SI) via the resazurin reduction assay. These protocols are based on the ability of resazurin (oxidized blue form) to be irreversibly reduced by enzymes in viable cells and generate a red fluorescent resorufin product and can be easily expanded to the investigation of the antimicrobial activity in other microorganisms.
被忽视的热带病引起了众多研究项目的科学关注，旨在实现对其有效控制或消除。在此过程中，需要更前沿的公共卫生政策和研究来发现源自天然产物的新型、更安全有效的药物。在此，我们描述了用于筛选天然产物衍生化合物以确定其抗利什曼原虫效力的方案。为此，通过细胞培养法评估特级初榨橄榄油中的总酚馏分（TPF）对细胞外前鞭毛体和细胞内无鞭毛体 spp. 形式的作用。本文的目的是描述一个逐步的程序，该程序可通过刃天青还原测定法轻松应用于准确估计50%抑制浓度（IC）、50%细胞毒性浓度（CC）和选择性指数（SI）。这些方案基于刃天青（氧化蓝色形式）在活细胞中被酶不可逆还原并生成红色荧光试卤灵产物的能力，并且可以轻松扩展到对其他微生物抗菌活性的研究。
