Augmentation of antioxidant system: Contribution to antimalarial activity of Clerodendrum violaceum leaf extract

  • Balogun, Elizabeth Abidemi ;
  • Zailani, Ahmed Hauwa ;
  • Adebayo, Joseph Oluwatope
  • Received : 2014.03.10
  • Accepted : 2014.11.17
  • Published : 2014.11.30


Reactive oxygen species are known to mediate various pathological conditions associated with malaria. In this study, the antioxidant potential of Clerodendrum violaceum leaf extracts, an indigenous antimalarial remedy, was evaluated. Total phenol, flavonoid, selenium, vitamins C and E contents of Clerodendrum violaceum leaf extracts were determined. The free radical scavenging activities of the extracts against DPPH, superoxide anion and hydrogen peroxide coupled with their reducing power were also evaluated in vitro. Moreover, responses of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) in a rodent malaria model to a 4-day administration of Clerodendrum violaceum leaf extracts were also evaluated. The methanolic extract was found to contain the highest amounts of antioxidant compounds/element and also demonstrated the highest free radical scavenging activity in vitro. The results showed a significant decrease (p < 0.05) in SOD and CAT activities with a concurrent significant (p < 0.05) increase in GPx and GR activities in both erythrocytes and liver of untreated Plasmodium berghei NK65-infected animals compared to the uninfected animals. The extracts were able to significantly increase (p < 0.05) SOD and CAT activities and significantly reduce (p < 0.05) GPx and GR activities in both the liver and erythrocytes compared to those observed in the untreated infected animals. The results suggest the augmentation of the antioxidant system as one of the possible mechanisms by which Clerodendrum violaceum extract ameliorates secondary effects of malaria infection, alongside its antiplasmodial effect in subjects.


Clerodendrum;antioxidant;Plasmodium;oxidative stress


  1. Adebayo JO, Yakubu MT, Egwim, CE, Owoyele BV, Enaibe BU. Effect of ethanolic extract of Khaya senegalensis stem bark on some biochemical parameters on rat Kidney. J Ethnopharmacol. 2003;88:69-72.
  2. AOAC. Official methods of Analysis of A.O.A.C International, George WL ed, (Washington, DC, USA; Association of Official Analytical Chemists), 2005.
  3. Aebi, HJ. Catalase. In: Methods of enzymatic analyses, Bergmeyen HV ed. (New York, USA: Academic Press), pp. 673-683, 1998.
  4. Ahmed I, Mehmood Z, Mohammad F. Screening of some Indian medicinal plants for their antimicrobial properties. J Ethnopharmacol. 1998;62:183-193.
  5. Alfin-Slater RB, Morris RS. Vitamin E and lipid peroxidation. Adv Lipid Res. 1963;1:183-210.
  6. Atamna H, Ginsburg H. Origin of reactive oxygen species in erythrocytes infected with Plasmodium falciparum. Mol Biochem Parasitol. 1993;61:231-241.
  7. Balakrishnan SD, Anuradha CV. Exercise, depletion of antioxidants and antioxidant manipulation. Cell Biochem Funct. 1998;16:269-275.<269::AID-CBF797>3.0.CO;2-B
  8. Balogun EA, Adebayo JO, Zailani AH, Kolawole OM, Ademowo OG. Activity of ethanolic extract of Clerodendrum violaceum leaves against Plasmodium berghei in mice. Agric Biol J N Am. 2010;1:307-312.
  9. Bannister J, Bannister W, Rotilio G. Aspects of the structure, function and applications of superoxide dismutase. CRC Crit Rev Biochem. 1987;22:111-118.
  10. Becker K, Tilley L, Vennerstrom JL, Roberts D, Rogerson S, Ginsburg H. Oxidative stress in malaria parasite infectederythrocytes:host parasite interactions. Int J Parasitol. 2004;34:163-189.
  11. Braganza JM, Scott P, Bilton G, Schofield D, Chaliner C, Shiel N, Hunt LP, Bottigheiri T. Evidence for early oxidative stress in acute pancreatitis: Clues for correction. Int J Pancreatol. 1995;17:69-81.
  12. Claudio F, Marcu VGL, Dulinea SPA, Emerson SL. The role of platelet and plasma markers of antioxidant status and oxidative stress in thrombocytopenia aming patients with vivax malaria. Mem Inst Oswaldo Cruz. 2008;103:517-521.
  13. Chanda S, Dave R. In vitro models for antioxidant activity evaluation and some medicinal plants possessing antioxidant properties: an overview. Afr J Microbiol Res. 2009;3:981-996.
  14. Chang CC, Yang MH, Chern JC. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Analysis. 2002;10:178-182.
  15. Chelikani P, Fita I, Loewen P. Diversity of structures and properties among catalases. Cell Mol Life Sci. 2004;61:192-208.
  16. Cook NC, Samman S. Flavonoids-Chemistry, metabolism, cardioprotective effects and dietary sources. J Nutr Biochem. 1996;7:66-76.
  17. Das BS, Nanda WK. Evidence for erythrocyte lipid peroxidation in acute falciparum malaria. Trans Roy Soc Trop Med Hyg. 1999;93:58-62.
  18. De Tullio MC. The Mystery of Vitamin C. Nature Education. 2010;3:48-51.
  19. Dreher I, Schmutzler C, Jakob F, Kohrle J. Expression of selenoproteins in various rat and human tissues in cell lines. J Trace Elements Med Biol. 1997;42:83-91.
  20. Farombi EO, Owoeye O. Antioxidative and chemopreventive properties properties of Vernonia amygdalina and Garcinia biflavonoid: A review. Int J Environ Res Public Health. 2011;8:2533-2555.
  21. Goldberg DM, Spooner RJ. Methods of enzymatic analysis. Vol. 3. Bergmeyen HV ed. 3rd ed. (Deerfield Beach, USA: Verlog Chemie), pp 258-265, 1983.
  22. Hasan SMR, Mokarram HMD, Akter R, Jamila M, Ehsanul HM, Rahman S. DPPH free radical scavenging activity of some Bangladesh medicinal plants. J Med Plants Res. 2009:3:875- 879. san%20et%20al.pdf
  23. Gornall AG, Bardawill CT, David MM. Determination of serum protein by means of biuret reaction. J Biol Chem. 1949;177:751-766.
  24. Gyamfi MA, Yonamire M, Aniya Y. Free radical scavenging action of medicinal herbs from Ghana: Thonningia sanguine on experimentally induced liver injuries. Gen Pharmacol. 1999;32:661-667.
  25. Halliwell B, Gutteridge, JM. The antioxidants of human extracellular fluids. Arch Biochem Biophys. 1990;280:1-8.
  26. Hayes J, Flanagan J, Jousey I. Glutathione transferases. Annu Rev Pharmacol Toxicol. 2005;45:51-58.
  27. Herrera E, Barbas C. Vitamin E: action, metabolism and perspectives. J Physiol Biochem. 2001;57:43-56.
  28. Hunt NH, Stocker R. Oxidative stress and the redox status of malaria-infected erythrocytes. Blood Cells. 1990;16:499-526.
  29. Jacob HS, Jandl JH. Effects of sulfhydryl inhibition on red blood cell. 3. Glutathione in the regulation of the hexose monophosphate pathway. J Biol Chem. 1966;241:4243-4250.
  30. Kulkarni AG, Suryakar AN, Sardeshmukh AS, Rathi DB. Studies on biochemical changes with special reference to oxidant and antioxidants in malaria patients. Indian J Clin Biochem. 2003;18:136-149.
  31. Marklund S, Marklund G. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superozide dismutase. Eur J Biochem. 1974;47:469-474.
  32. McCombie SC. Self-treatment for malaria: the evidence and methodological issues. Health Policy Plan. 2002;17:333-344.
  33. Ngaha EO, Akanji MA, Madusolomo MA. Studies on correlation between chloroquine - induced tissue damage and serum changes in rats. Experientia. 1989;45:143-146.
  34. Moon RC, McCormick DL, Mehta RG. Inhibition of carcinogenesis by retinoids. Cancer Res. 1983;43:2469-2475.
  35. Muller S. Redox and antioxidant systems of the malaria parasite Plasmodium falciparum. Mol Microbiol. 2004;53:1291-1305.
  36. Nabavi SM, Ebrahimzadeh MA, Nabavi SF, Hamidinia A, Bekhradnia AR. Determination of antioxidant activity, phenol and flavonoids content of Parrotia persica Mey. Pharmacologyonline. 2008;2:560-567.
  37. Noda Y, Anzai K, Mori A, Kohno M, Shimnei M, Packer L. Hydroxyl and superoxide anion radical scavenging activities of natural source antioxidants using the computerized JES FR30 ESR Spectrophotometer system. Biochem Mol Biol Inter. 1997;42:35-44.
  38. Padayatty S, Katz A, Wang Y, Eck P, Kwon O, Lee J, Chen, S, Corpe C, Dutta S, Levine M. Vitamin C as an antioxidant:Evaluation of its role in disease prevention. J Am Coll Nutr. 2003;22:18-35.
  39. Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med. 1967;70:158-169.
  40. Peters W. Drug resistance in Plasmodium berghei. Vinka and lips. Exp Parasitol. 1965;17:80-89.
  41. Postma NS, Mommers EC, Eling WMC, Zuidema J. Oxidative stress in malaria: Implications for prevention and therapy. Pharm World Sci. 1996;18:121-129.
  42. Rao NM. Medical Biochemistry (New Delhi, India: New Age International (P) Limited, Publishers), pp 565-568, 2006.
  43. Shimada K, Fujikawa K, Yahara K, Nakamura T. Antioxidative properties of xanthan on the autooxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem. 1992;40:945-948.
  44. Rice-Evans CA, Miller NJ, Paganga GC. Antioxidant properties of phenolic compounds. Trends Plant Sci. 1997;2:152-159.
  45. Sardesai VM. Role of antioxidants in health maintenance. Nutr Clin Pract. 1995;10:19-25.
  46. Shahidi F, Wanasundara PK. Phenolic antioxidants. Crit Rev Food Sci Nutr. 1992;32:67-103.
  47. Sohail M, Kaul A, Raziuddin M, Adak T. Decreased glutathione-S-transferase activity: diagnostic and protective role in vivax malaria. Clin Biochem. 2007;40:377-382.
  48. Spallhoiz JE, Boylan LM, Larsen HS. Advances in understanding Selenium role in the immune system. Ann N Y Acad Sci. 1990;587:123-139.
  49. Terashima K, Takaya Y, Niwa M. Powerful antioxidative agents based on garcinoic acid from Garcinia Kola. Bioorg Med Chem. 2002;10:1619-1635.
  50. Tona L, Mesia K, Ngimbi NP, Chrimwami B, Okond'ahoka Cimanga K, Bruyne T, Apers S, Hermans N, Totte J, Pieters L, Vlietinck AJ. In-vivo antimalarial activity of Cassia occidentalis. Morinda morindoides and Phyllanthus niruri. Annal Trop Med Parasitol. 2001;95:47-57.
  51. vander Jagt DL, Hunsaker LA, Campos NM, Scaletti JV. Localization and characterization of haemoglobin-degrading aspartic proteinases from the malarial parasite P. falciparum. Biochim Biophys Acta. 1992;1122:256-264.
  52. WHO. Promoting the role of traditional medicine in health systems: a strategy for the African region, WHO-Regional Office for Africa, Harare. 2001. Available at: (accessed on 9th March 2000)
  53. Yalcin O, Karatas F, Erulas FA, Ozdemir E. The levels of glutathione peroxidase, vitamin A, E, C and lipid peroxidation in patients with transitional cell carcinoma of the bladder. BJU Int. 2004;93:863-866.
  54. Yaping Z, Wenli Y, Dapu W, Xiaofeng L, Tianxi H. Chemiluminescence determination of free radical scavenging abilities of tea pigments and comparison with tea polyphenols. Food Chem. 2003;80:115-118.
  55. Yen GH, Chen HY. Antioxidant activity of various tea extracts in relation to their antimutagenicity. J Agric Food Chem. 1995; 43:27-32.
  56. Youdim KA, Dobbie MS, Kuhnle G, Proteggente AR, Abbott NJ, Rice-Evans C. Interaction between flavonoids and the blood brain barrier: in vitro studies. J Neurochem. 2003;85:180-192.
  57. Zelko IN, Mariani TJ, Folz RJ. Superoxide dismutase multigene family: A comparison of the CuZn-SOD (SOD 1), Mn-SOD (SOD 2) and ECSOD (SOD 3) gene structures, evolution and expression. Free Radic Biol Med. 2002;33:337-349.
  58. Zhao GR, Xiang ZJ, Ye TX, Yuan YJ, Guo ZX. Antioxidant activities of Salvia miltiorrhiza and Panax notoginseng. Food Chem. 2006;99 767-774.
  59. Zhou YC, Zheng RL. Phenolic compounds and an analog as superoxide anion scavenger and antioxidants. Biochem Pharm. 1991;42:1177-1199.


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