A Metabolomic Investigation of the Effect of Eosin B on Game-tocyte of Plasmodium falciparum Using 1HNMR Spectroscopy

BACKGROUND: Recently eosin B was shown to have an effect on the asexual stage of Plasmodium falciparum and in this study, its activity against gametocytes and changes in the culture medium metabolites were investigated using an(1)HNMR-based metabolomics approach. METHODS: In the Biochemistry Department of Pasteur Institute of Iran in 2017, parasites were cultured and gametocytogenesis induced by heparin and 5% hematocrit. Sexual stage parasites were tested by eosin B in 90 well plates and IC(50) determined using Lactate Dehydrogenase assay. Gametocytes were treated by IC(50) dose of eosin B and the medium collected in the two groups: with eosin B and controls and sent for (1)HNMR spectroscopy. The spectra were analyzed on MATLAB interface and the altered metabolites in the culture medium and eosin-affected biochemical pathways were identified by Human Metabolome Database and Metabo-analyst website. RESULTS: The results revealed eosin B had an effective gametocytocidal activity against P. falciparum. The significant metabolites changed in the medium were thia-mine, Asp, Asn, Tyr, Lys, Ala, Phenylpyruvic acid, NAD(+) and lipids. The main pathways identified were aminoacyl-tRNA biosynthesis, Phenylalanine, tyrosine and tryptophan biosynthesis, Alanine, aspartate and glutamate metabolism, Phenylala-nine metabolism, Nicotinate and nicotinamide metabolism, and lysine degradation. CONCLUSION: Eosin B exhibited substantial gametocytocidal activity and affected important drug targets in the Plasmodium.