Laboratory of Cellular Immunology, Department of Microbiology, Hellenic Pasteur Institute, 127 Vas. Sofias av. 11521 Athens, Greece.
Laboratory of Cellular Immunology, Department of Microbiology, Hellenic Pasteur Institute, 127 Vas. Sofias av. 11521 Athens, Greece; Division of Pharmacognosy and Natural Product Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis Zografou GR-15771 Athens, Greece.
Phytomedicine. 2018 Aug 1;47:143-150. doi: 10.1016/j.phymed.2018.04.030. Epub 2018 May 10.
Leishmaniasis is a neglected and emerging disease with varying clinical manifestations. The current treatment options rely on limited chemotherapy with serious drawbacks. Thus, there is an increasing interest in the identification of new candidates for designing potent, less toxic and low-cost drugs.
The purpose of this study was to evaluate the potential antileishmanial activity of the total phenolic fraction (TPF) derived from extra virgin olive oil (EVOO) when added in in vitro and in vivo experimental models of Leishmania infection.
We investigated the in vitro antileishmanial activity of TPF against two Leishmania species: a viscerotropic (L. infantum) and a dermotropic (L. major) strain. The antileishmanial effect was also tested in vivo in a murine cutaneous leishmaniasis model using L. major-infected BALB/c mice.
Separation and analytical methodologies were applied in order to extract the olive oil phenols (TPF) and determine the concentration of the major ones, respectively. The in vitro antileishmanial activity of TPF against promastigotes and intracellular amastigotes was determined by the resazurin cell viability assay. The TPF-induced nitric oxide synthesis by L. infantum and L. major -infected J774A.1 macrophages was determined using the Griess reaction, while the respective generation of reactive oxygen species was assessed by flow cytometry. Moreover, L. major-infected BALB/c mice were treated with TPF and its in vivo therapeutic effect was determined as reduction of the footpad swelling.
Our data showed that TPF exhibits inhibitory effect against cell free promastigotes and intracellular amastigotes of both L. infantum and L. major parasite strains. TPF demonstrated to be selectively active against Leishmania amastigotes and its antileishmanial activity was possibly mediated by reactive nitrogen and oxygen intermediates generated from the infected J774A.1 macrophages. Furthermore, administration of TPF in BALB/c mice infected with L. major caused significant reduction of footpad swelling demonstrating in vivo its antileishmanial effect. Based on HPLC-DAD analysis the major components of TPF are tyrosol, hydroxytyrosol, oleacein and oleocanthal.
This study brings a new low-cost candidate to the leishmaniasis drug discovery pipeline, upon further pharmacological investigation.
利什曼病是一种被忽视且不断出现的疾病，具有不同的临床表现。目前的治疗选择依赖于有限的化疗，但存在严重的缺陷。因此，人们越来越关注寻找新的候选药物，以设计出有效、毒性低且成本低的药物。
本研究旨在评估特级初榨橄榄油（EVOO）总酚类（TPF）在体外和体内利什曼原虫感染实验模型中的潜在抗利什曼活性。
我们研究了 TPF 对两种利什曼原虫的体外抗利什曼活性：一种内脏利什曼原虫（L. infantum）和一种皮肤利什曼原虫（L. major）株。还在感染 L. major 的 BALB/c 小鼠的皮肤利什曼病模型中进行了体内抗利什曼作用测试。
应用分离和分析方法分别提取橄榄油酚类（TPF）并确定主要成分的浓度。通过 Resazurin 细胞活力测定法测定 TPF 对前鞭毛体和细胞内无鞭毛体的体外抗利什曼活性。通过格里斯反应测定 TPF 诱导 L. infantum 和 L. major 感染的 J774A.1 巨噬细胞产生一氧化氮的情况，而通过流式细胞术评估各自的活性氧产生情况。此外，用 TPF 治疗感染 L. major 的 BALB/c 小鼠，并确定其体内治疗效果，即减少足垫肿胀。
我们的数据表明，TPF 对两种利什曼原虫（L. infantum 和 L. major）的无细胞前鞭毛体和细胞内无鞭毛体均具有抑制作用。TPF 对利什曼无鞭毛体具有选择性活性，其抗利什曼活性可能是由感染的 J774A.1 巨噬细胞产生的活性氮和氧中间体介导的。此外，在感染 L. major 的 BALB/c 小鼠中给予 TPF 可显著减少足垫肿胀，证明其体内抗利什曼作用。基于 HPLC-DAD 分析，TPF 的主要成分是酪醇、羟基酪醇、油酸和油酸辛酯。
在进一步的药理学研究基础上，本研究为利什曼病药物发现提供了一种新的低成本候选药物。
