Antiplasmodial, antioxidant and immunomodulatory activities of ethanol extract of Vernonia amygdalina del. Leaf in Swiss mice

Document Type : Original Research Article


1 Department of Biochemistry, Faculty of Life Sciences, University of Benin, PMB 1154, Benin City, Nigeria

2 In vivo Animal Testing Facility, Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow, India


Objective: Vernonia amygdalina (V.  amygdalina) leaf is locally employed in the Southern region of Nigeria in the treatment of malari
a infection. This study evaluated the in vivo antiplasmodial, antioxidant and immunomodulatory effect of ethanol extract of V.  amygdalina leaf.
Materials and Methods: The active principles of the dried leaf were extracted with ethanol. For quality validation, chemical finger-print of the extract was performed through high performance thin layer chromatography (HPTLC). The extract was assessed for antiplasmodial activity by the standard four-day suppressive test on Plasmodium berghei (ANKA) infected male Swiss mice (six weeks old) placed into five groups of six animals each.
Result: The absorption spectra from the HPTLC revealed several peaks suggesting presence of some bioactive compounds. Results from the in vivo study showed that the ethanol extract of the plant leaf was significantly active against P. berghei in a dose-dependent manner with the minimum and maximum activity observed in the mice treated orally with 100mg/kg (% inhibition of 23.7%) and 1000 mg/kg (% inhibition of 82.3 %) of the extract, respectively, on day four of the study. There was also a dose-dependent decrease (p<0.05) in some oxidative stress indices including nitric oxide and lipid peroxidation levels in the extract treated groups as against the non-treated infected group which had high levels of these parameters. The pro-inflammatory cytokines (TNF-α and IFN-ɣ) levels were also considerably low in the extract treated groups relative to the non-treated infected group.
Conclusion: The results suggest that ethanol extract of V. amygdalina leaf was active, with some immunomodulatory effect, against P. berghei infection.


Main Subjects

Abdulelah HAA, Zainal-Abidin BAH. 2007. In vivo anti-malarial tests of Nigella sativa (black seed) in different extracts. Am J Pharmacol Toxicol, 2: 46-50.
Abosi AO, Raseroka BH. 2003. In vivo antimalarial activity of Vernonia amgydalina. Br J Biomed Sci, 60: 89-91.
Adegbolagun OM, Emikpe BO, Woranola IOO, Ogunremi Y. 2014. Synergistic effect of aqueous extract of Telfaria occidentalis on the biological activities of artesunate in Plasmodium berghei infected mice. Afr Health Sci, 14: 111-118.
Akinpelu DA, 1999. Antimicrobial activity of Vernonia amygdalina leaves. Fitoterapia 70: 432-435.
Ayoola GA, Folawewo AD, Adesegun SA, Abioro OO, Adepoju-Bello AA, Coker HAB. 2008. Phytochemical and antioxidant screening of some plants of apocynaceae from South West, Nigeria. Afr J Plant Sci, 2: 124-128.
Atawa CB, Adamu AM, Gefu JO, Ajanusi OJ, Abdu PA, Chiezey AP, Atawa JN, Bowman DD. 2003. In vitro screening of 2 Nigerian medicinal plants (Vernonia amygdalina and Annona senegalensis) for anthelmintic activity. Vet Parasitol, 113: 73-81.
Awasthi A, Kumar A, Upadhyay SN, Yamada T, Matsunaga Y. 2003. Nitric oxide protects against chloroquine resistant Plasmodium yoeli nigeriensis parasites in vitro. Exp Parasitol, 105: 184-191.
Bashawri LAM, Mandil AA, Bahnassy AA, Ahmed MA. 2002. Malaria: Hematological aspects. Ann. Saudi Mediterr, 22: 372-377.
Brunet LR. 2001. Nitric acid in parasitic infections. Int Immunopharmacol, 1: 1457-1467.
Challand S, Willcox M. 2009. A clinical trial of the traditional medicine Vernonia amygdalina in the treatment of uncomplicated malaria. J Altern Com Med, 15: 1231-1237.
Chung I, Seo S, Kaug E, Park W, Moon H. 2009. Anti-malarial activity of 6-(8’Z-pentadecenyl)-salicyclic acid from Viola websteri in mice. Malarial J, 8: 151-154.
Clark IA, Budd AC, Alleva LM, Cowden WB. 2006. Human malarial disease: A consequence of inflammatory cytokine. Malarial J, 5: 85-116.
Das BS, Nanda NK. 1999. Evidence for erthrocyte lipid peroxidation in acute falciparum malaria. Trans Royal Soc Trop Med Hyg, 93: 58-62.
Depinay N, Franetich JF, Gruner AC, Mauduit M, Chavatte JM, Sauerwein RW, Siksik J, Hannoun L, Snounou G, Renia L, Luty AJF, Germert GV. 2011. Inhibitory effect of TNF-α in malarial pre-erythrocytic stage development: Influence of host hepatocyte/parasite combinations. PLOS One, 6: 17464-17472.
Dikasso D, Makonnen E, Debella A, Abebe D, Urga K, Makonnen W, Melaku D, Assefa A, Makonnen Y. 2006. In vivo anti-malarial activity of hydroalcoholic extracts from Asparagus africanus Lam. In mice infected with Plasmodium berghei. Ethiop J Health Dev, 20: 112-118.
Egwunyenga AO, Isamah G, Nmorsi OP. 2004. Lipid peroxidation and ascorbic acid levels in Nigeria children with acute falciparum malaria. Afr J Biotechnol, 3: 560-563.
Erasto P, Grierson DS, Afolayan AJ. 2006. Bioactive sesquiterpenes lactones from the leaves of Vernonia amygdalina. J Ethnopharmacol, 106: 117-120.
Erasto P, Grierson DS, Afolayan AJ. 2007. Evaluation of antioxidant activity and fatty acid profile of the leaves of Vernonia amygdalina growing in South Africa. Food Chem, 104: 636-642.
Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. 1982. Analysis of nitrate, nitrite and [15N] nitrate in biological fluids. Anal Biochem, 126: 131-138.
Golenser J, Chevion M. 1989. Oxidant species and malaria host parasite relationships in normal and abnormal erythrocytes. Seminars  Haematol, 26: 313-325.
Gutteridge JMC, Wilkins S. 1982. Copper-dependent hydroxyl radical damage to ascorbic acid: Formation of a thiobarbituric acid reactive product. Fed Eur Biomed Societies Letts, 327-330.
Heddini A. 2002. Malaria pathogenesis: A jigsaw with an increasing number of pieces. Int J Parasitol, 32: 1587-1598.
Kraft C, Jenett-Siems K, Siems K, Jakupovic J, Mavi S, Bienzle U, Eich E. 2003. In vitro antiplasmodial evaluation of medicinal plants from Zimbabwe. Phytother Res, 17: 123-128.
Kumari GN, Masilamani S, Ganesh MR, Aravind S, Sridhar SR. 2003. Zaluzanin D: a fungistatic sesquiterpenes from Vernonia arborea. Fitoterapia, 74: 479-482.
Omoregie ES, Osagie AU, 2011. Effect of J. tanjorensis leaves supplement on the activities of some antioxidant enzymes, vitamins and lipid peroxidation in rats. J Food Biochem, 35: 409-424.
Omoregie ES, Pal A, Sisodia B. 2011. In vitro antimalarial and cytotoxic activities of leaf extracts of Vernonia amygdalina (Del.). Nig J Basic Appl Sci, 19: 121-126.
Osamor VC, 2010. The etiology of malaria scourge: A comparative study of endemic nations of Africa and Asia. J Biol Sci, 10: 440-447.
Park W, Lee S, Moon H. 2008. Antimalarial activity of a new stilbene glycoside from Parthenocissus tricuspidata in mice. Antimicrob Agents  Chemother, 52: 3451-3453.
Peters W, Robinson BL. 1992. The chemotherapy of rodent malaria XLVII: studies on pyronaridine and other mannich base antimalarials. Ann Trop Med Parasitol, 86: 455-465.
Richards AL. 1997. Tumour necrosis factor and associated cytokines in the host’s response to malaria. Int J Parasitol, 27: 1251-1263.
Sinha S, Mishra SK, Sharma S, Patibandla PK, Mallick PK, Sharma SK, Mohanty S, Pati S, Mishra S, Ramteke BK, Bhatt RM, Joshi H, Dash AP, Ahuja RC, Awashi S., Venkatesh V, Habib S. 2008. Polymorphisms of TNF-enhancer and gene for Fcy RIIa correlate with the severity of falciparum malaria in the ethnically diverse Indian population. Malaria J, 7: 13-24.
Sobolewski P, Gramaglia I, Francgos J, Intaglietta M, Heyde HC. 2005. Nitric oxide bioavailability in malaria. Trends Parasitol, 21: 415-422.
Syamsudin T, Wahyuono S, Darmono T, Mustofa D. 2007. In vitro and in vivo antiplasmodial activities of stem bark extracts of Garcinia parvifolia Miq. (Guttiferae). Int J Trop Med, 2: 41-44.
Tona L, Cimanga RK, Mesia K, Musuamba CT, De Bruyne T, Apers S, Hernans N, Van Miert S, Pieters L, Totte J, Vlietinck AJ. 2004. In vitro antiplasmodial activity of extracts and fractions from seven medicinal plants used in the Democratic Republic of Congo. J Ethnopharmacol, 93: 27-32.
Torres D. 2009. Early production of ɣ-interferon in clinical malaria: Role of interleukin-18 and interleukin-12. Clin Infect Disease, 48: 1481-1482.
Trager W, Jensen JB. 1997. Continuous culture of Plasmodium falciparum: Its impact on malaria research. Int J Parasitol, 27: 989-1006.
WHO. 2005. The world malaria report from WHO and UNICEF. World Health Organisation, Geneva.
Wright CW. 2005. Traditional antimalarials and the development of novel antimalarial drugs. J Ethnopharmacol, 100: 67-71.
Yeo TW, Lampah DA, Tjitra E, Gitawati R, Kenangalem E, Piera K, Granger DL, Lopansri BK, Weinberg JB, Price RN, Duffull SB, Celermajer DS, Anstey NM. 2009. Relationship of cell-free hemoglobin to impaired endothelial nitric oxide bioavailability and perfusion in severe falciparum malaria. J Infect Disease, 200: 1522-1529.