Nefang is a polyherbal product composed of Mangifera indica (bark and leaf), Psidium guajava, Carica papaya, Cymbopogon citratus, Citrus sinensis, and Ocimum gratissimum (leaves), used for the treatment of malaria. Compounds with antioxidant activity are believed to modulate plasmodial infection. Antioxidant activity of the constituent aqueous plants extracts, in vitro, was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH), total phenolic content (TPC), and ferric reducing antioxidant power (FRAP) methods and, in vivo, Nefang (100 and 500？mg？kg？1) activity was evaluated in carbon tetrachloride-induced oxidative stressed Wistar rats. Superoxide dismutase, catalase activities, and lipid peroxidation by the malondialdehyde and total proteins assays were carried out. P. guajava, M. indica leaf, and bark extracts had the highest antioxidant properties in all three assays, with no statistically significant difference. Rats treated with the carbon tetrachloride had a statistically significant decrease in levels of triglycerides, superoxide dismutase, and catalase ( ) and increase in malondialdehyde activity, total protein levels, and liver and renal function markers, whereas rats treated with Nefang showed increased levels in the former and dose-dependent decrease towards normal levels in the later. These results reveal the constituent plants of Nefang that contribute to its in vivo antioxidant potential. This activity is a good indication of the therapeutic potential of Nefang. 1. Introduction Molecular oxygen is an indispensable element for the life of aerobic organisms because it enables the formation of reactive oxygen species (ROS) which in small quantities are essential for many physiological processes. At high doses, the ROS are very toxic [1]. The main ROS in humans are superoxide anion , hydrogen peroxide , radical hydroxide and the reactive nitrogen species (RNS), and nitric monoxide (NO). These are constantly generated within the cells upon exposure to xenobiotics, pollutants, ultraviolet rays, smoke, and some endogenous metabolites of the redox and respiratory chain during transfer of electrons [2]. Under normal physiological conditions, there is equilibrium between levels of intracellular ROS and the endogenous antioxidant system. The endogenous antioxidant system is composed of enzymes such as the superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione (GSH). Over production of ROS beyond the antioxidant capacity of the physiological system gives way to oxidative stress. Excessive oxidative stress leads to
References
[1]
M. Martínez-Cayuela, “Oxygen free radicals and human disease,” Biochimie, vol. 77, no. 3, pp. 147–161, 1995.
[2]
J. M. Borg and A. Reeber, Biochimie Métabolique, Ellipses, Paris, France, 2008.
[3]
B. Halliwell and J. M. C. Gutteridge, Free Radicals in Biology and Medicine, Oxford University Press, 4th edition, 2007.
[4]
J. L. Farber, “Mechanisms of cell injury by activated oxygen species,” Environmental Health Perspectives, vol. 102, no. 10, pp. 17–24, 1994.
[5]
K. Pong, “Oxidative stress in neurodegenerative diseases: therapeutic implications for superoxide dismutase mimetics,” Expert Opinion on Biological Therapy, vol. 3, no. 1, pp. 127–139, 2003.
[6]
M. Valko, C. J. Rhodes, J. Moncol, M. Izakovic, and M. Mazur, “Free radicals, metals and antioxidants in oxidative stress-induced cancer,” Chemico-Biological Interactions, vol. 160, no. 1, pp. 1–40, 2006.
[7]
V. B. Djordjevic, “Free radicals in cell biology,” International Review of Cytolology, vol. 237, pp. 57–89, 2004.
[8]
J. K. Willcox, S. L. Ash, and G. L. Catignani, “Antioxidants and prevention of chronic disease,” Critical Reviews in Food Science and Nutrition, vol. 44, no. 4, pp. 275–295, 2004.
[9]
F. L. Song, R. Y. Gan, Y. Zhang, Q. Xiao, L. Kuang, and H. B. Li, “Total phenolic contents and antioxidant capacities of selected chinese medicinal plants,” International Journal of Molecular Sciences, vol. 11, no. 6, pp. 2362–2372, 2010.
[10]
A. Meziti, Activité antioxydante des extraits des graines de Nigella sativa L: étude in vitro et in vivo. Mémoire réalisé en vue de l'obtention du Magister en Biochimie appliquée, Université de El Hadj Lakhdar, Batna, Algeria, 2009.
[11]
T. Finkel and N. J. Holbrook, “Oxidants, oxidative stress and the biology of ageing,” Nature, vol. 408, no. 6809, pp. 239–247, 2000.
[12]
G. A. Agbor, D. Kuate, and J. E. Oben, “Medicinal plants can be good source of antioxidants: case study in Cameroon,” Pakistan Journal of Biological Sciences, vol. 10, no. 4, pp. 537–544, 2007.
[13]
G. Cao, S. L. Booth, J. A. Sadowski, and R. L. Prior, “Increases in human plasma antioxidant capacity after consumption of controlled diets high in fruit and vegetables,” American Journal of Clinical Nutrition, vol. 68, no. 5, pp. 1081–1087, 1998.
[14]
S. S. Leonard, D. Cutler, M. Ding, V. Vallyathan, V. Castranova, and X. Shi, “Antioxidant properties of fruit and vegetable juices: more to the story than ascorbic acid,” Annals of Clinical and Laboratory Science, vol. 32, no. 2, pp. 193–200, 2002.
[15]
S. Naskar, A. Islam, U. K. Mazumder, P. Saha, P. K. Haldar, and M. Gupta, “In vitro and in vivo antioxidant potential of hydromethanolic extract of phoenix dactylifera fruits,” Journal Scientific Research, vol. 2, no. 1, pp. 144–157, 2010.
[16]
H. Sies, “Oxidative stress: an introduction,” in Oxidative Stress: Oxidants and Antioxidants, H. Sies, Ed., pp. 15–22, Academic Press, London, UK, 1991.
[17]
K. Becker, L. Tilley, J. L. Vennerstrom, D. Roberts, S. Rogerson, and H. Ginsburg, “Oxidative stress in malaria parasite-infected erythrocytes: host-parasite interactions,” International Journal for Parasitology, vol. 34, no. 2, pp. 163–189, 2004.
[18]
I. A. Clark and N. H. Hunt, “Evidence for reactive oxygen intermediates causing hemolysis and parasite death in malaria,” Infection and Immunity, vol. 39, no. 1, pp. 1–6, 1983.
[19]
P. G. Kremsner, B. Greve, B. Lell, D. Luckner, and D. Schmid, “Malarial anaemia in African children associated with high oxygen-radical production,” Lancet, vol. 355, no. 9197, pp. 40–41, 2000.
[20]
M. Guha, S. Kumar, V. Choubey, P. Maity, and U. Bandyopadhyay, “Apoptosis in liver during malaria: role of oxidative stress and implication of mitochondrial pathway,” FASEB Journal, vol. 20, no. 8, pp. E439–E449, 2006.
[21]
S. M. Potter, A. J. Mitchell, W. B. Cowden et al., “Phagocyte-derived reactive oxygen species do not influence the progression of murine blood-stage malaria infections,” Infection and Immunity, vol. 73, no. 8, pp. 4941–4947, 2005.
[22]
C. C. Keller, P. G. Kremsner, J. B. Hittner, M. A. Misukonis, J. B. Weinberg, and D. J. Perkins, “Elevated nitric oxide production in children with malarial anemia: hemozoin-induced nitric oxide synthase type 2 transcripts and nitric oxide in blood mononuclear cells,” Infection and Immunity, vol. 72, no. 8, pp. 4868–4873, 2004.
[23]
M. Sohail, A. Kaul, M. Raziuddin, and T. Adak, “Decreased glutathione-S-transferase activity: diagnostic and protective role in vivax malaria,” Clinical Biochemistry, vol. 40, no. 5-6, pp. 377–382, 2007.
[24]
D. G. Grahame-Smith and J. K. Aronson, Tratado de Farmacologia Clínica e Farmacoterapia, Guanabara Koogan, Rio de Janeiro, Brazil, 3rd edition, 2004.
[25]
S. Zhang, H. Chen, and G. S. Gerhard, “Heme synthesis increases artemisinin-induced radical formation and cytotoxicity that can be suppressed by superoxide scavengers,” Chemico-Biological Interactions, vol. 186, no. 1, pp. 30–35, 2010.
[26]
S. Percário, D. R. Moreira, B. A. Q. Gomes et al., “Oxidative stress in malaria,” International Journal of Molecular Sciences, vol. 13, no. 12, pp. 16346–16372, 2012.
[27]
P. A. Reis, C. M. Comim, F. Hermani et al., “Cognitive dysfunction is sustained after rescue therapy in experimental cerebral malaria, and is reduced by additive antioxidant therapy,” PLoS Pathogens, vol. 6, no. 6, Article ID e1000963, 2010.
[28]
P. A. Tarkang, F. A. Okalebo, G. A. Agbor, N. Tsabang, A .N. Guantai, and G. M. Rukunga, “Indigenous Knowledge and folk use of a polyherbal antimalarial by the Bayang Community, South West Region of Cameroon,” Journal of Natural Product and Plant Resources, vol. 2, no. 3, pp. 372–380, 2012.
[29]
National Research Council of the National Academies, Guide for Care and Use of Laboratory Animals, National Academies Press, Washington, DC, USA, 8th edition, 2010.
[30]
S. S. Handa, S. P. S. Khanuja, G. Longo, and D. D. Rakesh, Extraction Technologies for Medicinal and Aromatic Plants, ICS-UNIDO International Centre for Science and High Technology, Trieste, Italy, 2008.
[31]
G. C. Yen and P. D. Duh, “Scavenging effect of methanolic extracts of peanut hulls on free-radical and active-oxygen species,” Journal of Agricultural and Food Chemistry, vol. 42, no. 3, pp. 629–632, 1994.
[32]
K. Slinkard and V. L. Singleton, “Total phenol analysis: automation and comparism with manual method,” American Journal of Enology and Viticulture, vol. 28, pp. 49–455, 1977.
[33]
I. F. F. Benzie and J. J. Strain, “The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay,” Analytical Biochemistry, vol. 239, no. 1, pp. 70–76, 1996.
[34]
H. P. Misra and I. Fridovich, “The generation of superoxide radical during the autoxidation of hemoglobin,” Journal of Biological Chemistry, vol. 247, no. 21, pp. 6960–6962, 1972.
[35]
A. K. Sinha, “Colorimetry of catalase,” Analytical Biochemistry, vol. 11, pp. 469–474, 1972.
[36]
T. Biswas, J. K. Pal, K. Naskar, D. K. Ghosh, and J. Ghosal, “Lipid peroxidation of erythrocytes during anemia of the hamsters infected with Leishmania donovani,” Molecular and Cellular Biochemistry, vol. 146, no. 2, pp. 99–105, 1995.
[37]
O. H. Lowry, N. J. Rosebrouch, A. L. Farr, and R. J. Randall, “Protein measurement with the folin phenol reagent,” The Journal of Biological Chemistry, vol. 193, no. 1, pp. 265–275, 1951.
[38]
A. Kinoshita, Y. Nakayama, T. Kitayama, and M. Tomita, “Simulation study of methemoglobin reduction in erythrocytes: differential contributions of two pathways to tolerance to oxidative stress,” FEBS Journal, vol. 274, no. 6, pp. 1449–1458, 2007.
[39]
M. Wang, Y. Jin, and C. T. Ho, “Evaluation of resveratrol derivatives as potential antioxidants and identification of a reaction product of resveratrol and 2,2-diphenyl-1-picryhydrazyl radical,” Journal of Agricultural and Food Chemistry, vol. 47, no. 10, pp. 3974–3977, 1999.
[40]
L. P. Leong and G. Shui, “An investigation of antioxidant capacity of fruits in Singapore markets,” Food Chemistry, vol. 76, no. 1, pp. 69–75, 2002.
[41]
M. Hu and L. H. Skibsted, “Antioxidative capacity of rhizome extract and rhizome knot extract of edible lotus (Nelumbo nuficera),” Food Chemistry, vol. 76, no. 3, pp. 327–333, 2002.
[42]
R. L. Prior, X. Wu, and K. Schaich, “Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements,” Journal of Agricultural and Food Chemistry, vol. 53, no. 10, pp. 4290–4302, 2005.
[43]
H. Tsukamoto, M. Matsuoka, and S. W. French, “Experimental models of hepatic fibrosis: a review,” Seminars in Liver Disease, vol. 10, no. 1, pp. 56–65, 1990.
[44]
J. Terblanche and R. Hickman, “Animal models of fulminant hepatic failure,” Digestive Diseases and Sciences, vol. 36, no. 6, pp. 770–774, 1991.
[45]
S. Mizuguchi, S. Takemura, Y. Minamiyama et al., “S-allyl cysteine attenuated CCl4-induced oxidative stress and pulmonary fibrosis in rats,” BioFactors, vol. 26, no. 1, pp. 81–92, 2006.
[46]
K. C. Preethi and R. Kuttan, “Hepato and reno protective action of calendula officinalis L. Flower extract,” Indian Journal of Experimental Biology, vol. 47, no. 3, pp. 163–168, 2009.
[47]
P. H. Stern, T. Furukawa, and T. M. Broody, “Rat liver and plasma lipids after carbon tetrachloride administration,” Journal of Lipid Research, vol. 6, pp. 278–286, 1965.
[48]
W. L. Hubbell and H. M. McConnell, “Molecular motion in spin-labeled phospholipids and membranes,” Journal of the American Chemical Society, vol. 93, no. 2, pp. 314–326, 1971.
[49]
R. Anandan, R. Deepa Rekha, and T. Devaki, “Protective effect of Picrorhiza kurroa on mitochondrial glutathione antioxidant system in D-galactosamine-induced hepatitis in rats,” Current Science, vol. 76, no. 12, pp. 1543–1545, 1999.
[50]
J. Eritsland, “Safety considerations of polyunsaturated fatty acids,” American Journal of Clinical Nutrition, vol. 71, supplement 1, pp. 197S–201S, 2000.
[51]
B. M. Cavanaugh, Nurse’s Manual of Laboratory and Diagnostics Tests, F. A. Davis Company, Philadelphia, Pa, USA, 4th edition, 2003.
