In vitro Antisickling properties, Free radicals quenching potential, protective properties against oxidative mediated ion toxicity of combinations of Theobroma cocoa beans (sterculaceae)

Prosper Biapa, Floriane Tammo, Jesse Juenkou Tekumu, Prudence Nya Nkwikeu, Natacha Lena Yembeau, Christian Fotsing Kengne, Phelix Bruno Telefo, Anatole Constant Pieme


Sickle cell anaemia induces an abnormal haemoglobin synthesis and reactive oxygen species. The present work aims to determine the antioxidant and antisickling activities of Theobroma cocoa extracts combination. Cocoa beans Extracts combinations of Santchou/Obala (CSOH), Bertoua/Mbalmayo (CMBH) were prepared using water/ethanol mixture. The antioxidant capacity of extracts was determined by measuring FRAP, DPPH° and ABTS°. The protective activity of extracts was estimated by evaluating some oxidative stress parameters. Osmotic fragility resistance and antisickling properties of the combination were investigated. CSOH and CMBH respectively revealed a high content of polyphenols, flavonoids. Extracts scavenged DPPH° and ABTS° radicals, respectively with IC50 values between 4.07 μg/mL and 13.15 μg/mL. CMBH showed the best antisickling reduction activity (54.56% and 70.07% after 2h30 and 24h respectively). The haemolysis percentage of erythrocytes decreased with an increase of the extract concentrations. Extracts combinations decreased and increased respectively the MDA and GSH levels as well as normalizing the enzymatic activities. Extracts combinations of cocoa have antioxidants and antisickling properties and could be used as a potential agent against sickle cell anaemia.


Sickle cell anemia, oxidative stress, antioxidant, antisickling,plants

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ORPHANET. Encyclopédie Orphanet Grand Public. La Drépanocytose. Tiré de Pub/fr/Drepanocytose-FRfrPub125v01.pdf. 2011; Consulté le 16 Avril 2017.

Catonné Y.; Mukisimukasa M.; Rouvillain J.; Ribeyre D. Manifestations ostéo-articulaires de la drépanocytose. Maîtrise Orthopédique. 2004; pp. 135.

Hebbel R.; Morgan W.; Eaton J. Accelerated autoxidation and heme loss due to instability of sickle haemoglobin. Proceeding of the National Academy Sciences USA 85 (1988) 237-241.

Aubry P.; Gaüzère B. Hémoglobinoses actualité. Médecine Tropicale. 2014; Consulté le 16 Mai 2017.

DREPAVIE. Intégrer le problème de santé publique de la drépanocytose lors de l'EPU du 2013; Consulté le 16 Avril 2017.

Markus S.A.; Oliver S.; Ayse Hulya O.B.; Gabriele M.B. La drépanocytose en Suisse, 2e partie: Mesures thérapeutiques et prophylactiques. Revue médicale Suisse 8(34) (2008) 606–608.

Steinberg M. Pathophysiologically based drug treatment of sickle cell disease. Trends in Pharmacological Sciences 27(4) (2006) 204-210.

Gbadamosi I.; Adeyemi S.; Adeyemi A.; Moody J. In vitro antisickling activities of two indigenous plant recipes in Ibadan. International Journal of Phytomedecines 4 (2012) 205-211.

Mpiana P.; Misakabu F.; Tshibangu D.; Ngbolua K.; Mwanangombo D. Antisickling activity and membrane stabilizing effect of anthocyanins extracts from Adansonia digitata L. barks on sickle blood cells. International Blood Research & Reviews 2(5) (2014) 198-212.

Nanfack P.; Biapa N.; Pieme C.; Ama M.; Moukette B.; Ngogang Y. The in vitro antisickling and antioxidant effects of aqueous extract Zanthoxyllum heitti on sickle cell disorder. BMC Complement Alternative Medicine 13(162) (2013) 1-7.

N’draman-Donou E.; Fofié Y.; Adjambri E.; Mélèdje M.F.; Sawadogo D. Caractérisation et évaluation in vitro de l’effet antifalcémiant des graines de Cajanus cajan (Fabacées) sur les drépanocytes à Abidjan-Cote d’ Ivoire. International Journal of Biological and Chemical Sciences 9(5) (2015) 2300-2308.

Nguelewou F. Evaluation in vitro des effets antifalcémiants et antioxydants des extraits hydro-éthanoliques de Theobroma cacao et du chocolat noir. Mémoire de Master soutenue à l’Université de Dschang, 2016.

Yembeau L.N. Evaluation in vitro des propriétés antifalcémiantes et antioxydantes des extraits hydro-éthanoliques de Theobroma cacao des régions de l’Est et du Sud du Cameroun. Mémoire de Master soutenue à l’Université de Dschang , 2017.

Benhammou N.; Bekkara F.; Panovska T. Biological activities of the essential oil and ethanolic extract of Inula viscosa from the Tlemcen region of Algeria. Advances in Food Sciences 30 (3) (2008) 132-139.

Benzie I.; Strain J. The ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: the FRAP assay. Analytical Biochemistry 239 (1996) 70-76.

Molyneux P. The use of the stable free radical diphenylpicryl-hydrazyl (DPPH) for estimating antioxidant activity. Songklanakarin Journal of Science and Technology 26 (2) (2004) 212-219.

Re R.; Pellegrini N.; Proteggente A.; Pannala A.; Yang M.; Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine‎ 26 (1999) 1231-1237.

Jaja S.I.; Kehinde M.O.; Ghenebitse S.; Mojiminyi A.I. Effect of vitamin C on arterial blood pressure, irreversible sickled cells and osmotic fragility in sickle cell anaemia subjects. Nigerian journal of physiological sciences16 (1-2) (2000) 14-18.

Gornall A.; Bardawill C.; David M. Determination of serum proteins by means of the biuret reaction. Journal of Biological Chemistry 177(2) (1949) 751-766.

Wilbur K.; Bernheim F.; Shapiro O. Determination of lipid peroxidation. Archives of Biochemistry and Biophysics 24 (1949) 305-310.

Ellman G. Tissue sulphydryl groups. Archives of Biochemistry and Biophysics 82 (1959) 70-77.

Sinha A. Colorimetric assay of catalase. Analytical Biochemistry 47 (1972) 389–394.

Misra H.; Fridovich I. The role of superoxyde dismutase anion in the oxydation of epinephrine and a simple assay for superoxide dismutase. Journal of Biological Chemistry 247(10) (1972) 3170-3175.

Singleton V.; Orthofer R.; Lamuela R. Analysis of total and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods in Enzymology 299 (1999) 152-178.

Zhishen J.; Mengcheng T.; Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry 64 (4) (1999) 555– 559.

Luczkiewicz M.; Cisowski W.; Kaiser P.; Ochocki R.; Piotrowski A. Comparative analysis of phenolic acids in mistletoe plants from various hosts. Acta poloniae pharmaceutica 58 (2001) 373-379.

Vicas S.; Rugina D.; Sosaciu C. Antioxidant activities of Viscum album’s leaves from various host trees. Bulletin of University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca 65 (2008) 327-332.

Thilagam E.; Kumarappan C.; Parimaladevi B.; Mandal S. Antioxidant Activity of Senna surattensis leaves. International Journal of advances in Pharmacy, Biology and Chemistry 1 (2012) 454–460.

Bahorun T.; Luximon-Ramma A.; Crozier A.; Aruoma O. Total phenol, flavonoid, proanthocyanidins and vitamin C levels and antioxidant activities of Mauritian vegetables.‎ Journal of the Science of Food and Agriculture 84 (2004) 1553-1561.

Adeolu A.; Florence O.; Anthony J.; Patrick J. Antioxidant activities and phenolic contents of the methanol extracts of the stems of Acokanthera oppositifolia and Adenia gummifera. BMC Complementary and Alternative Medicine 8 (2008) 1-7.

Bidié A.; N’Guessan B.; Yapo A.; N’Guessan J. ; Djaman A. Activités antioxydantes de dix plantes médicinales de la pharmacopée ivoirienne. Sciences Nat 8 (1) (2011) 1-11.

Messaadia A. L’effet protecteur d’un antioxydant naturel contre le stress oxydatif induit par le nitrate d’ammonium chez les rats. Thèse en vue de l’obtention d’un Diplôme de Doctorat 3ème Cycle, Université badjimokhtar – annaba, 2014.

Santos S.; Flores P.; Valadez B.; Fernandez R.; Martinez V.; Salas C. Polyphenolic content, free radical scavenging activity and isolation of Tiliroside from Heliocarpus terebinthinaceus (Tiliaceae) Seeds. Journal of Biological Sciences 14 (5): (2014) 376-380.

Mukherjee S.; Pawar N.; Kulkarni O.; Nagarkar B.; Thopte S.; Bhujbal A.; et al. Evaluation of free-radical quenching properties of standard Ayurvedic formulation Vayasthapana rasayana. BMC Complementary and Alternative Medicine 11 (2011) 11:38.

Seck M.; Sall C.; Gueye P.; Seck I.; Dioum M.; Lembachar Z.; Gueye R.; Fall D.; Fall M.; Dieye T. Etude de l’activité antifalcémiante d’extraits de racines de Leptadenia hastate decne. (Asclepiadacae). International Journal of Biological and Chemical Sciences 9(3) (2015) 1375-1383.

Egunyomi A.; Moody J.; Eletu O. Antisickling activities of two ethnomedicinal plant recipes used for the management of sickle cell anaemia in Ibadan, Nigeria. African Journal of Biotechnology 8 (1) (2009) 020-025.

Ngbolua K. Evaluation de l’activité antidrepanocytaire et antipaludique de quelques taxons végétaux de la RDC et de Madagascar. Thèse de Doctorat, Université de Kinshasa, 2012.



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