The Effect of Aqueous Extract of Cinnamon on the Metabolome ofPlasmodium falciparumUsing¹HNMR Spectroscopy

Abstract

Malaria is responsible for estimated 584,000 deaths in 2013. Researchers are working on new drugs and medicinal herbs due to drug resistance that is a major problem facing them; the search is on for new medicinal herbs. Cinnamon is the bark of a tree with reported antiparasitic effects. Metabonomics is the simultaneous study of all the metabolites in biological fluids, cells, and tissues detected by high throughput technology. It was decided to determine the mechanism of the effect of aqueous extract of cinnamon on the metabolome of Plasmodium falciparum in vitro using 1HNMR spectroscopy. Prepared aqueous extract of cinnamon was added to a culture of Plasmodium falciparum 3D7 and its 50% inhibitory concentration determined, and, after collection, their metabolites were extracted and 1HNMR spectroscopy by NOESY method was done. The spectra were analyzed by chemometric methods. The differentiating metabolites were identified using Human Metabolome Database and the metabolic cycles identified by Metaboanalyst. 50% inhibitory concentration of cinnamon on Plasmodium falciparum was 1.25 mg/mL with p < 0.001. The metabolites were identified as succinic acid, glutathione, L-aspartic acid, beta-alanine, and 2-methylbutyryl glycine. The main metabolic cycles detected were alanine and aspartame and glutamate pathway and pantothenate and coenzyme A biosynthesis and lysine biosynthesis and glutathione metabolism, which are all important as drug targets.

Influential parameters for the analysis of intracellular parasite metabolomics

“…Despite implementing current best practices, including erythrocyte lysis and washing steps to remove parasites from their intracellular milieu ( Fig. 1A, (3,8,12,22)), parasite separation from the host is poor. Microscopy revealed that parasites lysed from host cells remain embedded in erythrocyte membranes and washes fail to isolate parasite material ( Fig.…”

“…We found that extra-parasite material contributed by host erythrocytes and media components can also heavily impact the metabolome. Many studies employ erythrocyte lysis prior to sample purification ( (3,8,12,22), see Materials and Methods). However, several novel results from our study show that this step does not eliminate the potential for host contamination.…”

“…Additionally, the measurement of direct mediators of the phenotype, such as signaling and biosynthetic metabolites, can improve the ability to characterize phenotypes mediated by proteins that are not yet annotated into the genome. For this reason, metabolomics is becoming increasingly popular in studies of intra-erythrocytic stages of P. falciparum (3)(4)(5)(6)(7)(8)(9)(10)(11)(12). These studies have improved our understanding of malaria pathogenesis (13), strain-specific phenotypes (11), and host-parasite interactions (9).…”

Phytochemical Characterization and Chemotherapeutic Potential of Cinnamomum verum Extracts on the Multiplication of Protozoan Parasites In Vitro and In Vivo

“…C. verum has several medicinal properties, such as anti-inflammatory, hypoglycemic, antibacterial, antioxidant, spasmolytic, antidiarrheal, antifungal, antitumor, analgesic, gastroprotective, anticancer, and anthelmintic ones [14,19,20]. Interestingly, recent studies have documented the antimalarial efficacy of C. verum extract [21,22]. However, to our knowledge, no data has been reported to show the efficacy of C. verum herbal extracts against the growth of piroplasm parasites.…”

Cholesterol-dependent enrichment of understudied erythrocytic stages of humanPlasmodiumparasites

“…The P. falciparum genome is poorly annotated (40% of genes) compared to many model organisms (85% of genes for S. cerevisiae), making metabolomics an attractive method for unbiased study of parasite biology (7,8). Indeed, Plasmodium metabolomics has become increasingly popular in recent years for investigation of parasite biology and antimalarial drug action (9)(10)(11)(12)(13)(14)(15)(16)(17)(18).…”