In Vitro Biological Activities of Drepanoalpha® Ethanolic Extract, A Justicia Secunda and Moringa Oleifera-Based Phytomedicine Proposed for The Symptomatic Treatment of Sickle Cell Disease

Benjamin Gbolo Zoawe; Amandine Nachtergael; Damien S. T. Tshibangu; Nicole M. Misengabu; Victoire Nsabatien; Patrick B. Memvanga; Dorothée D. Tshilanda; Jean-Paul K. N. Ngbolua; Pius T. Mpiana; Pierre Duez

doi:10.18196/jfaps.v3i2.16000

In Vitro Biological Activities of Drepanoalpha® Ethanolic Extract, A Justicia Secunda and Moringa Oleifera-Based Phytomedicine Proposed for The Symptomatic Treatment of Sickle Cell Disease

Benjamin Gbolo Zoawe, Amandine Nachtergael, Damien S. T. Tshibangu, Nicole M. Misengabu, Victoire Nsabatien, Patrick B. Memvanga, Dorothée D. Tshilanda, Jean-Paul K. N. Ngbolua, Pius T. Mpiana, Pierre Duez

Abstract

Sickle cell disease (SCD) is an autosomal recessive blood disorder characterized by red blood cells that assume an abnormal, rigid sickle shape under low-oxygen conditions. These sickle-shaped erythrocytes tend to lyse, aggregate, and obstruct small blood vessels, leading to major complications. The present study aims to investigate properties that may underlie the activity of Drepanoalpha^â, an antisickling herbal formulation developed in the Democratic Republic of Congo (DRC) for the prevention and symptomatic treatment of sickle cell disease crises. The Drepanoalpha^® Ethanolic Extract (DEE) is a dry extract (drug-extract ratio, DER, 100/11) prepared from ethanol (96 %, v/v) percolation of a 1:1 mixture of 2 food plants, Justicia secunda Vahl and Moringa oleifera Lam. Sickling was classically measured by light microscopy on diluted washed erythrocytes obtained from homozygote patients; erythrocytes were treated with 2 % Na₂S₂O₅ in the presence of DEE (suspension in 9 ‰ NaCl), 9 ‰ NaCl (negative control) or disodium cromoglycate (DSCG, positive control). For all tested conditions, the sickle hemoglobin polymerization, the Fe²⁺/Fe³⁺ ratio, and the median corpuscular fragility were measured by spectrophotometry. The DEE reversed sickling by 89.1 %, comparable to DSCG (87.7 %; 60.3 µg/mL), inhibiting sickle cell hemoglobin polymerization of 77.8 % and 74.4 %, respectively. The Fe²⁺/Fe³⁺ ratio was improved by 18.0 % for DEE and 15.9 % for DSCG. The median corpuscular fragility values were 0.602, 0.714, and 0.732 for NaCl 9 ‰, DSCG, and DEE, respectively. The measured in vitro parameters validate an effective antisickling effect of DEE and confirm the value of this improved traditional herbal formulation for the management of SCD.

Keywords

antisickling activity; drepanoalpha®; erythrocytes; Fe2+/Fe3+ ratio; hemoglobin polymerization; sickle cell disease

Full Text:

PDF

References

Gbolo BZ, Asamboa LS, Bongo GN, Tshibangu DST, Kasali F, Memvanga P, et al. Bioactivity and Chemical Analysis of Drepanoalpha : An Anti-Sickle Cell Anemia Poly-Herbal Formula from Congo-Kinshasa. Am J Phytomedicine Clin Ther. 2017;5(1):1–7. https://doi.org/10.9734/JOCAMR/2017/37350

Aufradet E. Drépanocytose et activité physique : conséquences sur les mécanismes impliqués dans l’adhérence vasculaire, l’inflammation et le stress-oxydatif. Thèse en vue de l’obtention d’un doctorat en Sciences et techniques des activités physiques et sportives, Université de Lyon, France. [Lyon]: University of Lyon; 2012.

Imaga NA. Phytomedicines and nutraceuticals: Alternative therapeutics for sickle cell anemia. Sci World J. 2013;2013:1–12. https://doi.org/10.1155/2013/269659

Okoh MP, Alli LA, Tolvanen MEE, Nwegbu MM. Herbal Drug use in Sickle Cell Disease Management; Trends and Perspectives in Sub-Saharan Africa - A Systematic Review. Curr Drug Discov Technol. 2019;16(4):372–85. https://doi.org/10.2174/1570163815666181002101611

Verlhac S, Kandem A, Bernaudin F, Vasile M. L’accident vasculaire cerebral chez l’enfant drepanocytaire. Efficacite du protocole de prevention par doppler transcranien. J Radiol. 2007;88(10):1453. https://doi.org/10.1016/S0221-0363(07)81397-7

Shongo MY a. P, Mukuku O, Lubala TK asol., Mutombo AM ulang., Kanteng GW akam., Umumbu WS ombod., et al. Sickle cell disease in stationary phase in 6-59 months children in Lubumbashi: epidemiology and clinical features. Pan Afr Med J. 2014;19(71):1–7.

Tshilolo L, Aissi LM, Lukusa D, Kinsiama C, Wembonyama S, Gulbis B, et al. Neonatal screening for sickle cell anaemia in the Democratic Republic of the Congo : experience from a pioneer project on 31 204 newborns. J Clin Pathol. 2009;62:35–8. https://doi.org/10.1136/jcp.2008.058958

Abere TA, Egharevba CO, Chukwurah IO. Pharmacognostic evaluation and antisickling activity of the leaves of Securinega virosa Roxb. ex Willd. ( Euphorbiaceae). African J Biotechnol. 2014;13(40):4040–5.

Joseph Kahumba, Tsiry Rasamiravaka, Philippe Ndjolo Okusa, Amuri Bakari, Léonidas Bizumukama, Jean-Baptiste Kalonji, Martin Kiendrebeogo, Christian Rabemenantsoa, Mondher El Jaziri, Elizabeth M. Williamson PD. Traditional African medicine: from ancestral know-how to bright future. Science (80- ). 2015;350(6262):871–871.

WHO. WHO Traditional Medicine Strategy: 2014-2023. WHO (World Heal Organ ) Libr Cat Data, Geneva. 2013;78.

Manya MH, Keymeulen F, Ngezahayo J, Bakari AS, Mutombo EK, Kahumba BJ, et al. Antimalarial herbal remedies of Bukavu and Uvira areas in DR Congo: An ethnobotanical survey. J Ethnopharmacol. 2020;249:1–28. https://doi.org/10.1016/j.jep.2019.112422

Mahavy CE, Duez P, Eljaziri M, Rasamiravaka T. African Plant-Based Natural Products with Antivirulence Activities to the Rescue of Antibiotics. Antibiotics. 2020;9:1–30. https://doi.org/10.3390/antibiotics9110830

Chanda S, Parekh J, Vaghasiya Y, Dave R, Baravalia Y, Nair R. Medicinal Plants - From Traditional Use to Toxicity Assessment:A Review: Int J Pharm Sci Res. 2015;6(7):2652–70.

Mpiana PT, Ngbolua K-N, Tshibangu STD. Les alicaments et la drépanocytose : une mini-revue. Comptes Rendus Chim. 2016;19:884–9. https://doi.org/10.1016/j.crci.2016.02.019

Ngbolua K, Mpiana PT. The Possible Role of a Congolese polyherbal formula ( Drepanoalpha ) as source of Epigenetic Modulators in Sickle Cell Disease : A Hypothesis. J Adv Med Life Sci Res. 2014;2(1):1–3.

Kitadi JM, Lengbiye EM, Gbolo BZ, Inkoto CL, Muanyishay CL, Lufuluabo GL, et al. Justicia secunda Vahl species : Phytochemistry, Pharmacology and Future Directions : a mini-review. Discov Phytomedicine. 2019;6(4):157–71. https://doi.org/10.15562/phytomedicine.2019.93

Koffi NG, Henri KK, Djakalia O. Plants used to treat anaemia , in traditional medicine , by Abbey and Krobou populations , in the South of Côte-d ’ Ivoire. J Appl Sci Res. 2010;6(8):1291–7.

Mpiana PT, Bokota MT, Tshibangu DST, Ngbolua KN, Atibu EK, Kwembe JTK, et al. Antisickling activity of three species of justicia from Kisangani ( DR Congo ): Int J Biol Chem Sci [Internet]. 2010;4(6):1953–61.

Ngbolua K-N. An Updated review on the Bioactivities and Phytochemistry of the Nutraceutical Plant Moringa oleifera Lam (Moringaceae) as valuable phytomedicine of multi-purpose. Discov Phytomedicine. 2018;5(4):52–63. https://doi.org/10.15562/phytomedicine.2018.71

Leone A, Spada A, Battezzati A, Schiraldi A, Aristil J, Bertoli S. Cultivation, genetic, ethnopharmacology, phytochemistry and pharmacology of Moringa oleifera leaves: An overview. Int J Mol Sci. 2015;16(6):12791–835. https://doi.org/10.3390/ijms160612791

Maldini M, Maksoud SA, Natella F, Montoro P, Petretto GL, Foddai M, et al. Moringa oleifera: Study of phenolics and glucosinolates by mass spectrometry. J Mass Spectrom. 2014;49(9):900–10. https://doi.org/10.1002/jms.3437

Tshingani K, Donnen P, Mukumbi H, Duez P, Dramaix-Wilmet M. Impact of Moringa oleifera lam. Leaf powder supplementation versus nutritional counseling on the body mass index and immune response of HIV patients on antiretroviral therapy: A single-blind randomized control trial. BMC Complement Altern Med. 2017;17(1):1–13. https://doi.org/10.1186/s12906-017-1920-z

Shah SK, Jhade DN, Chouksey R. Moringa oleifera Lam. A study of ethnobotany, nutrients and pharmacological profile. Res J Pharm Biol Chem Sci. 2016;7(5):2158–65.

Nwaoguikpe R, Ujowundu C, Igwe C, Dike P. The Effects of Moringa oleifera Leaves Extracts on Sickle Cell Hemoglobin. J Sci Res Reports. 2015;4(2):123–32. https://doi.org/10.9734/JSRR/2015/12905

MPiana PT, Misakabu FM, Kitadi JM, Ngbolua KN, Tshibangu DST, Lombe BK, et al. Antisickling activity and physico-chemical stability of anthocyanin extracts from Hypoxis angustifolia Lam (Hypoxidaceae) Bulbs. In: Nohuru Motohashi, editor. Occurrences, Structure, Biosynthesis, and Health Benefits Based on Their Evidences of Medicinal Phytochemicals in Vegetables and Fruits. 3rd ed. Yew York: Nova Science Publishers,Inc; 2014. p. 97–114.

Bennett RN, Mellon FA, Foidl N, Pratt JH, Dupont MS, Perkins L, et al. Profiling glucosinolates and phenolics in vegetative and reproductive tissues of the multi-purpose trees Moringa oleifera L. (Horseradish tree) and Moringa stenopetala L. J Agric Food Chem. 2003;51(12):3546–53. https://doi.org/10.1021/jf0211480

COE (Council of Europe). High-performance thin-layer chromatography of herbal drugs and herbal drug preparations (2.8.25). 10th ed. EDQM, editor. Vol. 1, European Pharmacopoeia. Strasbourg Cedex: EDQM; 2019. 4370 p.

Kotue TC, Djote WNB, Marlyne M, Pieme AC, Kansci G, Fokou E. Antisickling and Antioxidant Properties of Omega-3 Fatty Acids EPA/DHA. Nutr Food Sci Int J [Internet]. 2019;9(1):555752. Available from: https://juniperpublishers.com/nfsij/NFSIJ.MS.ID.555752.php

Noguchi CT, Schechter AN. Inhibition of Sickle Hemoglobin Gelation by Amino Acids and Related Compounds. Biochemistry. 1978;17(25):5455–9. https://doi.org/10.1021/bi00618a020

Bizumukama L, Ferster A, Gulbis B, Kumps A, Cotton F. In vitro inhibitory effects of disodium cromoglycate on ionic transports involved in sickle cell dehydration. Pharmacology. 2009;83(5):318–22. https://doi.org/10.1159/000215598

Brugnara C, De Franceschi L. New therapeutic approaches to sickle cell disease. Hématologie. 2006;12(4):239–45.

Ngbolua KN, Mudogo V, Mpiana PT, Malekani MJ, Herintsoa, Rafatro, Ratsimamanga, S. Urverg, Takoy, L, Rakotoarimana, H and Tshibangu DST. Evaluation de l ’ activité anti-drépanocytaire et antipaludique de quelques taxons végétaux de la République démocratique du Congo et de Madagascar. Ethnopharmacologia. 2013;50:7–12.

Ravelojaona M. Analyse histologique des répercussions musculaires , structurales , énergétiques et microvasculaires chez des hommes et des femmes drépanocytaires. Thèse de Sciences Présentée et soutenue publiquement pour l’obtention du diplôme de Doctorat en Biologie Médecine Santé, Spécialité : Biologie et Physiologie de l’Exercice. UNIVERSITÉ JEAN MONNET SAINT ETIENNE, France. [Saint-Etienne]: Université Jean Monnet - Saint-Etienne; 2014.

Mpiana PT, Mudogo V, Tshibangu DS., Ngbolua KN, Shetonde OM, Mangwala PK, et al. In vitro antisickling activity of anthocyanins extract of a congolese plant: Alchornea cordifolia. M. Arg. J Med Sci. 2007;7(7):1182–6. https://doi.org/10.3923/jms.2007.1182.1186

Mpiana PT, Tshibangu DST, Shetonde OM, Ngbolua KN. In vitro antidrepanocytary actvity (anti-sickle cell anemia) of some congolese plants. Phytomedicine. 2007;14(2–3):192–5. https://doi.org/10.1016/j.phymed.2006.05.008

Mpiana PT, Mudogo V, Nyamangombe L, Kakule MK, Ngbolua KN, Atibu EK, et al. Antisickling activity and photodegradation effect of anthocyanins extracts from Alchornea cordifolia ( Schumach & Thonn ) and Crotalaria retusa. Ann Africaines Med. 2009;2(6):239–45.

Mpiana PT, Tshibangu DS, Ngbolua K, Tshilanda DD, Atibu EK. Antisickling Activity of Anthocyanins of Jatropha curcas L. In: Plants RP in M, editor. Chemistry and Medicinal Value. RPMP; 2007. p. 101–5.

Nwaoguikpe R, Braide W. The antisickling effects of some micronutrients and antioxidant vitamins in sickle cell disease management. J Med Med Sci. 2012;3(5):334–40. https://doi.org/10.4314/ijbcs.v3i5.51079

Ngbolua KN. Evaluation de l’activité anti-drépanocytaire et anti-paludique de quelques plantes de la République Démocratique du Congo et écotypes de Madagascar. Thèse présentée et soutenue pour obtenir le grade de Docteur en Sciences. Université de Kinshasa, RDC. Kinshasa; 2012.

4Mpiana PT, Ngbolua KTNN, Bokota MT, Kasonga TK, Atibu EK, Tshibangu DST, et al. In vitro effects of anthocyanin extracts from Justicia secunda Vahl on the solubility of haemoglobin S and membrane stability of sickle erythrocytes. Blood Transfus. 2010;8(4):248–54.

Abdullahi B. in Vitro Anti-Sickling Effect of Crude and Partially Purified Fractions of Methanolic Extract of Steculia Setigera Leaf on Human Sickled Red Blood Cells. Sci World J. 2018;13(4):81–6.

Richard AM, Diaz JH, Kaye AD. Reexamining the risks of drinking-water nitrates on public health. Ochsner J [Internet]. 2014;14(3):392–8. Available from: http://www.ncbi.nlm.nih.gov/pubmed/25249806%0Ahttp://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=PMC4171798

Nanfack P, Biapa N, Pieme C, Ama-Moor V, Moukette B, Yonkeu JN. The in vitro antisickling and antioxidant effects of aqueous extracts Zanthoxyllum heitzii on sickle cell disorder. BMC Complement Altern Med. 2013;13(162):1–7. https://doi.org/10.1186/1472-6882-13-162

Kiefer I, Prock P, Lawrence C, Bayer P, Rathmanner T, Kunze M, et al. Supplementation with Mixed Fruit and Vegetable Juice Concentrates Increased Serum Antioxidants and Folate in Healthy Adults. J Am Coll Nutr. 2004;23(3):205–11. https://doi.org/10.1080/07315724.2004.10719362

Tomc J, Debeljak N. Molecular pathways involved in the development of congenital erythrocytosis. Genes (Basel). 2021;12(1150):1–20. https://doi.org/10.3390/genes12081150

Kambale J, Ngolua K, Mpiana P, Mudogo V, Tshibangu D, Wumba D, et al. Evaluation in vitro de l’activité antifalcémiante et effet antioxydant des extraits d’Uapaca heudelotii Baill. (Euphorbiaceae). Int J Biol Chem Sci. 2013;7(2):523–34. https://doi.org/10.4314/ijbcs.v7i2.9

Mpiana PT, Misakabu FM, Tshibangu DST, Ngbolua KN, D.T. M. Antisickling Activity and Membrane Stabilizing Effect of Anthocyanins Extracts Antisickling Activity and Membrane Stabilizing Effect of Anthocyanins Extracts from Adansonia digitata L . Barks on Sickle. Int Blood Res Rev. 2014;2(5):198–212. https://doi.org/10.9734/IBRR/2014/10539

Gbolo ZB, Tshibangu STD, Memvanga BP, Bongo NG, Kasali MF, Ngbolua KN, et al. Assessment of the Efficacy and Tolerance of Drepanoalpha® in the Management of Sickle Cell Disease in Kinshasa (DR Congo): About Ten Cases. Int J Med Pharm Case Reports. 2017;9(2):1–10. https://doi.org/10.9734/IJMPCR/2017/33658

Gbolo ZB, Tshibangu D, Asamboa L, Bongo G, Kasali F, Feza V, et al. Sickle Cell Anemia Therapeutic Approach Based on Drepanoalpha ® : About 34 Sickle Cell Anemia Therapeutic Approach Based on Drepanoalpha ® : About 34 Cases. J Complement Altern Med Res. 2017;4(2):1–8. https://doi.org/10.9734/JOCAMR/2017/37350

DOI: https://doi.org/10.18196/jfaps.v3i2.16000