The Effect of (-)-Epigallocatechin 3-O - Gallate In Vitro and In Vivo in Leishmania braziliensis: Involvement of Reactive Oxygen Species as a Mechanism of Action

Abstract

BackgroundLeishmaniasis is a parasitic disease associated with extensive mortality and morbidity. The treatment for leishmaniasis is currently based on pentavalent antimonials and amphotericin B; however, these drugs result in numerous adverse side effects. Natural compounds have been used as novel treatments for parasitic diseases. In this paper, we evaluated the effect of (-)-epigallocatechin 3-O-gallate (EGCG) on Leishmania braziliensis in vitro and in vivo and described the mechanism of EGCG action against L. braziliensis promastigotes and intracellular amastigotes.Methodology/Principal Finding In vitro activity and reactive oxygen species (ROS) measurements were determined during the promastigote and intracellular amastigote life stages. The effect of EGCG on mitochondrial membrane potential (ΔΨm) was assayed using JC-1, and intracellular ATP concentrations were measured using a luciferin-luciferase system. The in vivo experiments were performed in infected BALB/c mice orally treated with EGCG. EGCG reduced promastigote viability and the infection index in a time- and dose-dependent manner, with IC50 values of 278.8 µM and 3.4 µM, respectively, at 72 h and a selectivity index of 149.5. In addition, EGCG induced ROS production in the promastigote and intracellular amastigote, and the effects were reversed by polyethylene glycol (PEG)-catalase. Additionally, EGCG reduced ΔΨm, thereby decreasing intracellular ATP concentrations in promastigotes. Furthermore, EGCG treatment was also effective in vivo, demonstrating oral bioavailability and reduced parasitic loads without altering serological toxicity markers.Conclusions/SignificanceIn conclusion, our study demonstrates the leishmanicidal effects of EGCG against the two forms of L. braziliensis, the promastigote and amastigote. In addition, EGCG promotes ROS production as a part of its mechanism of action, resulting in decreased ΔΨm and reduced intracellular ATP concentrations. These actions ultimately culminate in parasite death. Furthermore, our data suggest that EGCG is orally effective in the treatment of L. braziliensis-infected BALB/c mice without altering serological toxicity markers.

Effect of Apigenin on Leishmania amazonensis Is Associated with Reactive Oxygen Species Production Followed by Mitochondrial Dysfunction

“…[10][11][12]20 Treatment with 1 for 24 h increased ROS generation in L. amazonensis in a concentration-dependent manner (p < 0.01) ( Figure 2). ROS levels in L. amazonensis treated with 1 (96 μM) were 2.9-fold higher than those of the control.…”

Unifying Mechanism for Nutrients as Anticancer Agents: Electron Transfer, Reactive Oxygen Species and Oxidative Stress

“…The compound enhanced the production of radicals, both ROS and RNS (Jeong, Park, Cho et al, 2015). A study was made of the drug effect on Leishmania (Inacio, Gervazoni, Canto-Cavalheiro et al, 2014). Results suggest a mode of action involving ROS.…”

Novel arylalkylamine compounds exhibits potent selective antiparasitic activity against Leishmania major

“…19,20 A recent study suggested that biological compounds that involve the production of reactive oxygen species (ROS) may induce cell death in various leishmaniasis. 21,22 Almeida-Amaral and co-workers reported that the flavonol quercetin and the epigallocatechin gallate (Scheme 1. ), known potent antioxidants, induced death in Leishamania amazonensis and braziliensis respectively.…”

“…), known potent antioxidants, induced death in Leishamania amazonensis and braziliensis respectively. 21,22 Recently, Stockwell and co-workers reported the discovery of an arylalkylamine AA1-type compound (Fer-1) that was shown to be a potent and specific inhibitor of a unique form of non-apoptoptic, iron-dependent, oxidative cell death, known as ferroptosis. 23,24 The Fer-1 compound inhibits the lipid ROS production through an anti-oxidant mechanism.…”

“…We hypothesized, based on the recent study 21,22 that AA9 compound may induce cell death in L-Major through the production of ROS. We then investigated the intracellular ROS in L-major after treatment with AA9 compound.…”