Author/Authors :
Hajialiani, Fateme Medical Parasitology Department - School of Medicine -International Campus - Iran University of Medical Sciences, Tehran , Elmi, Taher Parasitology and Mycology Department - Faculty of Medicine - Iran University of Medical Sciences, Tehran , Mohamadi, Maryam Biochemistry Department - Pasteur Institute of Iran - Pasteur Avenue, Tehran , Sadeghi, Sedigheh Biochemistry Department - Pasteur Institute of Iran - Pasteur Avenue, Tehran , Shahbazzadeh, Delavar Laboratory of Venom and Bio Therapeutics Molecules - Department of Medical Biotechnology - Biotechnology Research Center - Pasteur Institute of Iran, Tehran , Ghaffarifar, Fatemeh Department of Parasitology - Faculty of Medical Sciences - Tarbiat modares University Tehran , Dalimi, Abdolhossein Department of Parasitology - Faculty of Medical Sciences - Tarbiat modares University Tehran , Arjmand, Mohammad Biochemistry Department - Pasteur Institute of Iran - Pasteur Avenue, Tehran , Tabatabaie, Fatemeh Parasitology and Mycology Department - Faculty of Medicine - Iran University of Medical Sciences, Tehran , Zamani, Zahra Biochemistry Department - Pasteur Institute of Iran - Pasteur Avenue, Tehran
Abstract :
Objective(s): Malaria is an important parasitic disease with high morbidity and mortality in tropical
areas. Resistance to most antimalarial drugs has encouraged the development of new drugs including
natural products. Venom is a complex mixture of active pharmaceutical ingredients. The purpose of
this study was to investigate the antimalarial activity of purified fractions of Naja naja oxiana.
Materials and Methods: Lyophilized venom was purified with a Sephacryl S-200 HR column and the
fractions lyophilized and inhibitory concentration 50% against Plasmodium falciparum 3D7 in vitro
obtained. The 4th fraction was run on a Mono Q column, and activity against P. falciparum was detected by
lactate dehydrogenase assay and purity by SDS PAGE. Large scale culture of the parasite was carried out
with and without the active fraction on the ring stage for 48 hr. The parasites were collected and lyophilized
and analyzed by 1HNMR. Chemometrics studies were performed using MATLAB, differentiating metabolites
were identified by Human Metabolic Database, and metabolic pathways by the Metaboanalyst online
package.
Results: The active fraction from the ion exchange column had a 50% inhibitory concentration of
0.026 μg/ml on P. falciparum in vitro (P<0.001) with molecular weight of 63 kDa by SDS-PAGE and no
hemolytic activity. Metabolomics studies on the two groups with and without the fraction identified 5
differentiating metabolites and a number of related pathways.
Conclusion: The metabolites were succinic acid, l-glutamic acid, pyruvic acid, cholesterol, and NAD. The
changes in the Krebs cycle and metabolism pathways of nicotinamide and pyruvate were noticeable.
Keywords :
Chemometrics , 1HNMR , Metabolomics , Naja naja oxiana venom , Plasmodium falciparum
