Full Length Research Paper
ABSTRACT
This study was designed to investigate the ability of ethanol fruit extract from Detarium microcarpum to protect erythrocytes against hemolysis and lipid peroxidation. To achieve this objective, hemolytic, anti-hemolytic and lipid peroxidation from cell membrane assays were used. Hemolytic and anti-hemolytic activities (regarding H2O2 induced hemolysis) were assessed by determination of free haemoglobin at 540 nm. Inhibition of lipid peroxidation was measured at 532 nm, using thiobarbituric acid reaction on sodium nitroprusside and ferric sulfate induced liposome peroxidation models. D. microcarpum fruit ethanol extract (100 µg/ml) did not exhibit any hemolytic effect but reduces significantly hemolysis from human and rat erythrocyte with inhibitory percentages more than 50 and 75%, respectively. Furthermore, the extract caused a significant decrease in both ferric sulfate and sodium nitroprusside inducing lipid peroxidation in each rat tissue liposomes investigated. D. microcarpum fruit ethanol extract protects erythrocytes against hemolysis and lipid peroxidation, probably due to its antioxidant potential. Therefore, animal tissues disorders caused by cell membrane lipid damage could be potentially managed/prevented by dietary intake of D. microcarpum fruit pulp.
Key words: Detarium microcarpum, anti-hemolytic activity, inhibition of lipid peroxidation
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
This study data demonstrated that fruit ethanol extract from D. microcarpum was an interesting radical fighter and seems benefitial in preventing cell membrane against oxidative damage. In the in vitro iron ion induced erythrocyte hemolysis model, hydroxyl radical is formed by hydrogen peroxide via Fenton reaction. This radical initiate the lipid peroxidation of bio membrane by electrophile attack, and the propagation of this peroxidation is conducted by alkoxyl and peroxyl radicals engendered, leading to haemoglobin liberation. Inhibition of hydrogen peroxide induced erythrocyte hemolysis by extract could be explained by the properties of extract to scavenge directly hydrogen peroxide by turning it into water molecule (James and Alewo, 2014)or to inhibit hydroxyl radicals formation via Fenton reaction, impeaching lipid peroxidation initiation (Su et al., 2009). Fruit extract could also stop lipid peroxidation propagation by alkoxyl and peroxyl radical scavenging. Verstraeten et al. (2004)reported in liposomes stability study, that flavonoids such as flavanols and related procyanidins can prevent the ferrous iron-mediated increase in membrane permeability. Together, these finding supported the hypothesis that these flavonoids can interact with the polar head group of lipids and consequently limit the incorporation of certain deleterious molecules that could affect membrane integrity and function. These finding suggested that the antihemolitic property of D. microcarpum fruit may be due in part to it richest in flavonoids content (Lamien-Meda et al., 2008). Rat erythrocytes were more protected than human erythrocytes against hydrogen peroxide toxicity according with previous studies (Pekiner, 2002). This author reported that membrane of rat erythrocyte was richer in lecithin contents than human erythrocyte, but the last one contained most rate of sphingomyelin. These finding suggested that sphingomyelin is more sensible to lipid peroxidation than lecithin. Regarding rat erythrocyte hemolysis test, extract showed higher erythrocyte protection than ascorbic acid. This finding suggested that ascorbic acid in certain conditions can exercise adverse effects on cell membrane stability. So, ascorbic acid in the presence of transition metals can react in turn to form radical, leading to cell membrane lipid peroxidation contributing to attenuate its membrane protective effect.
In iron ion and sodium nitroprusside induced liposomes lipid peroxidation model, iron ion can initiate liposomes lipid peroxidation by a direct one electron transfer on an unsaturated site in lipid carbon skeleton. Iron ion can also cause indirectly lipid peroxidation by hydroxyl radical formation via Fenton reaction and also accelerates peroxidation by decomposing lipid hydroperoxides into peroxyl and alkoxyl radicals that can themselves abstract hydrogen and perpetuate the chain reaction of lipid peroxidation (Ojo et al., 2014). In the in vitro sodium nitroprusside induced lipid peroxidation model, the hydrolysis of sodium nitroprusside in physiological middle releases Fe2+ causing lipid peroxidation via Fenton Reaction and the concomitant formation of oxide nitric radical can react together with other reactive oxygen species amplifying lipid peroxidation by secondary radical species formation (Akomolafe et al., 2012). The protected activity of extract against these two pro-oxidants could be explained by the properties of extract to chelate iron ion or to trap radical species formed by these pro-oxidants. Previously, demonstrated radicals scavenging effect of fruits from D. microcarpum could justify its lipid peroxidation inhibition properties (Lamien-Meda et al., 2008). Extract could also stop the propagation of lipid peroxidation by alkoxyl and peroxyl radicals’ neutralization. Extract inhibited more lipid peroxidation induced by iron ion than sodium nitroprusside (Figure 2a and b). This finding may be due to the probable synergic effect of Fe2+ and nitric oxide released from the hydrolysis of sodium nitroprusside on lipid peroxidation mediation (Khan, 2014). The highest sensibility of liver and brain to these pro-oxidants toxicity may be due to the presence of abundant polyunsaturated fatty acids in these organs. So, brain and liver contains higher amount of polyunsaturated fatty than heart, pancreas and kidney justifying their vulnerability to lipid peroxidation (De et al., 2008). These two organs are equally, potential sources of reactive oxygen species with a considerable reduction of antioxydant level comparatively to the other organs exposing their liposomes to lipid peroxidation (De et al., 2008). The use of many drugs is limited because of their cytotoxicity effects associated with cell membrane degradation and these actual results could encourage the dietary and medicinal intake of D. microcarpum fruit.
CONCLUSION
ACKNOWLEDGEMENTS
CONFLICT OF INTERESTS
REFERENCES
Akah PA, Nworu CS, Mbaoji FN, Nwabunike IA, Onyeto CA (2012). Genus Detarium: ethnomedicinal, phytochemical and pharmacological profile. Phytopharmacology 3:367-375. |
|
Akomolafe SF, Oboh G, Akindahunsi AA, Akinyemi AJ, Adeyanju O (2012). Inhibitory effect of aqueous extract of Moringa oleifera and Newbuoldia laevis leaves on ferrous sulphate and sodium nitroprusside induced oxidative stress in rat's tested in Vitro. ISRN Pharmacology 2013:119-128. |
|
Badmus JA, Odunola OA, Yekeen TA, Gbadegesin AM, Fatoki JO, Godo MO, Oyebanjo KS, Hiss DC (2013). Evaluation of antioxidant, antimutagenic, and lipid peroxidation inhibitory activities of selected fractions of Holarrhena floribunda (G. Don) leaves. Acta Biochimica Polonica 60:435-442. |
|
Bamisaye FA, Ajani EO, Nurain IO, Adebisi KE, Quadri RT, dMinari JB (2014). Evaluation of growth performance of rats fed with sweet detar, Detarium microcarpum fruit as supplementary feed ingredient. IOSR J. Environ. Sci. Toxicol. Food Technol. 8:115-121. |
|
Cavin A, Hay A, Marston A, Stoeckli-evans H, Scopelliti R, Diallo D, Hostettmann K (2006). Bioactive diterpenes from the fruits of Detarium microcarpum. J. Nat. Prod. 69:768-773. |
|
De S, Adhikari S, Tilak-Jain J, Menon VP, Devasagayam TPA (2008). Antioxidant activity of an aminothiazole compound: Possible mechanisms. Chem. Biol. Interact. 173:215-223. |
|
EEC (1986). Council Directive 86/609/EEC of 24 Novembre 1986 on the approximation of laws, regulation and administrative provisions of the member States regarding the protection of animals used for experimental and other Scientific purposes. Official J. Eur. Com. 358:1-29. |
|
James O, Alewo IM (2014). In vitro Antihemolytic Activity of Gymnema Sylvestre Extracts against Hydrogen Peroxide (H2O2) Induced Haemolysis in Human Erythrocytes. Am. J. Phytomed. Clin. Therapeut. 2:861-869. |
|
Khan A, Sabir SM, Nazar H, Hamid A, Khan U, Hussain A (2014). Antioxidant activities and inhibitory effects of dietary plants against sodium nitroprusside induced lipid peroxidation in the mouse brain and liver. Food Sci. Biotechnol. 23:1305-1311. |
|
Lamien-Meda A, Lamien CE, Compaoré MYM, Meda NTR, Kiendrebeogo M, Zeba B, Millogo JF, Nacoulma OG (2008). Polyphenol content and antioxidant activity of fourteen wild edible fruits from Burkina Faso. Molecules 13:581-594. |
|
Lima PPG, Vianello F, Corrêa CR, Arnoux R, Campos DS, Borguini MG (2014). Polyphenols in Fruits and Vegetables and Its Effect on Human Health. Food Nutr. Sci. 5:1065-1082. |
|
Obun CO, Yahaya SM, Kibon A, Ukim C (2010). Evaluation of Detarium microcarpum pulp meal as feed ingredient in the diets of growing rabbits. Am. J. Food. Nutr. 6:1822-1827. |
|
Ojo OA, Oloyede O, Tugbobo O, Olarewaju O, Ojo A (2014). Antioxidant and inhibitory effect of scent leaf (Ocimum gratissimum) on Fe2+ and sodium nitroprusside induced lipid peroxidation in rat Brain in vitro. Adv. Biol. Res. 8:8-17. |
|
Pekiner DB (2002). Fatty acid composition of red blood cell membrane phosphatidylethanolamine and phosphatidyletcholine in rat, rabbit, human and dog. J. Fac. Pharm. Ankara 31:169-182. |
|
Rauchová H, Vokurková M, Koudelová J (2012). Hypoxia-Induced Lipid Peroxidation in the Brain During Postnatal Ontogenesis. Physiol. Res. 61:89-101. |
|
Sasikumar JM, Maheshu V, Smilin AG, Gincy MM, Joji C (2012). Antioxidant and antihemolytic activities of common Nilgiri barberry (Berberis tinctoria lesch.) from south India. Int. Food Res. J. 19:1601-1607. |
|
Seifried HE, Anderson DE, Fisher EI, Milner JA (2007). A review of the interaction among dietary antioxidants and reactive oxygen species. J. Nutr. Biochem. 18:567-579. |
|
Su XY, Wang ZY, Liu JR (2009). In vitro and in vivo antioxidant activity of Pinus koraiensis seed extract containing phenolic compounds. Food Chem. 117: 681-686. |
|
Sumathy R, Sankaranarayanan S, Bama P, Ramachandran J, Vijayalakshmi M, Deecaraman M (2013). Antioxidant and antihemolytic activity of flavonoid extract from fruit peel of Punica granatum. Asian J. Pharm. Clin. Res. 6:211-214. |
|
Verstraeten SV, Oteiza PI, Fraga CG (2004). Membrane effects of Cocoa procyanidins in liposomes and jurkat T cells. Biol. Res. 37:293-300. |
|
Wahedi JA, David LD (2013). Effect of Detarium microcarpum fruit pulp on the haematological indices of rats in Mubi Adamawa state, Nigeria. Global J. Biol. Agric. Health Sci. 2:76-80. |
|
World Medical Association Declaration of Helsink (2013). Ethical Principles for Medical Research Involving Human Subjects. J. Am. Med. Assoc. 310(20):2191-2194. |
Copyright © 2024 Author(s) retain the copyright of this article.
This article is published under the terms of the Creative Commons Attribution License 4.0