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Background  Leishmaniasis 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 (ΔΨ m ) 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 IC 50  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 ΔΨ m , 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/Significance  In 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 ΔΨ m  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.

plos neglected tropical diseases

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