HEPATO CURATIVE AND ANTIOXIDANT EFFECT OF ETHYL-ACETATE AND N-BUTANOL FRACTIONS OF Detarium microcarpum STEM BARK IN CCl4 INDUCED LIVER DAMAGE IN WISTAR RATS
ABSTRACT
The stem bark of Detarium microcarpum (Guill and Perr.) is used in traditional medicine for the treatment of liver disease in middle belt region of Nigeria. To substantiate this folkloric claim, ethyl-acetate and n-butanol fractions of Detarium microcarpum stem bark was investigated for its hepatocurative and antioxidant effect in CCl4 induced liver damage in rats. Aqueous extraction was carried out on Detarium microcarpum stem bark and the crude extract was further fractionated sequentially using ethyl-acetate and n-butanol solvents. In the in-vitro studies, phytochemical screening of the crude extract showed the presence of phenolic, flavonoids, tannins, saponins, alkaloids and glycosides while total phenolic content assay, total flavonoid content assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH), Reducing power and H2O2 free radical scavenging activities were carried out on ethyl-acetate and n-butanol fractions. The total phenol content for n-butanol and ethyl acetate fractions were 2.97±0.31 and 11.54±0.20 mg/g Gallic acid equivalents while total flavonoid content were 234.42±0.71 and 45.76±2.59 mg/g quercetin equivalents. Ethyl acetate fraction showed the highest DPPH free radical scavenging activity with 65.31% inhibition while n-butanol showed the highest reducing power and H2O2 free radical scavenging activities with 65.31% and 52.55% which informed the choice of n-butanol fraction for further studies. In the in-vivo studies, the LD50 of n-butanol fraction of Detarium microcarpum stem bark was >5000 mg/kg body weight of rats. CCl4 (1ml/kg body weight) as a 1:1(v/v) solution in olive oil was used to induce liver damage followed by subsequent treatment with n-butanol fraction of Detarium microcarpum stem bark at three different doses (100, 150 and 200 mg/kg bw/day) while silymarin (100 mg/kg bw/day) was used as standard drug for 28 days. The liver weight was significantly (p<0.05) increased in the negative control group when compared with the CCl4 treated groups. There was significant (p<0.05) reduction in the serum activities of alanine aminotransaminase (ALT), aspartate aminotransaminase (AST), alkaline phosphatase (ALP), direct and indirect bilirubin for CCl4 treated groups compared to the negative control group. Total protein (TP) and albumin (ALB) in the negative control group were reduced but not significantly (p>0.05) compared to the CCl4 treated groups. In endogenous antioxidant activities, there was significant (p<0.05) reduction of malondialdehyde (MDA) in CCl4 treated groups compared to the negative control group. A significant (p<0.05) increase was also observed in superoxide dismutase (SOD) and catalase (CAT) activities of CCl4 treated groups compared to the negative control group. These results may suggest hepato curative and antioxidant effects of Detarium microcarpum stem bark in CCl4 induced liver damaged animals.
CHAPTER ONE
INTRODUCTION
1.1 Preamble
Herbal medicines are herbal preparations produced by subjecting plant materials to extraction, fractionation, purification, concentration or other physical or biological processes which may be produced for immediate consumption or as a basis for herbal products (WHO, 2001). Notwithstanding the extent of significant advancement in modern medicine in recent decades, plants still make an important contribution to health care. Traditionally they are used worldwide for the prevention and treatment of disease. Herbal plants were prescribed even when their active compounds were unknown because of their effectiveness and relatively low cost (Bhawna and Kumar, 2010). This observation is particularly more relevant to people in the developing countries of the world where the majority of the populations are living in the rural areas.
The liver plays an important role in regulating various physiological processes. It is essential in the body for maintenance, performance and regulating homeostatic functions. It is involved with almost all the biochemical pathways for growth, fight against diseases, nutrient supply, energy provision and reproduction. In addition, it aids metabolism of carbohydrate, protein and fat, detoxification, secretion of bile and storage of vitamins (Ahsan et al., 2009). Because of its central role in drug metabolism, it is the most vulnerable tissue for drug toxicity (Sunil et al., 2012). The role played by the liver in the removal of substances from the portal circulation makes it susceptible to persistent attack by offending foreign compounds, culminating in liver dysfunction (Bodkhe and Ram, 2007). The liver secretes bile, prothrombin, fibrinogen, blood-clotting factors and heparin, a mucopolysaccharide sulfuric acid ester that prevents blood from clotting within the circulatory system (Bhawna and Kumar, 2010). Toxic chemicals, xenobiotics, alcohol consumption, malnutrition, anaemia, medications, autoimmune disorders (Marina, 2006), viral infections (hepatitis A, B, C, D, etc.) and microbial infections (Sharma and Ahuja, 1997) are harmful and cause damage to the hepatocytes.
Reactive oxygen species (ROS) are continuously generated during metabolic processes to regulate a number of physiological functions essential to the body (Valko et al., 2007). These reactive oxygen species are prone to withdraw electrons from biological macromolecules such as proteins, lipids, nucleic acids in order to gain stability in the biological system. This disruption may be attributed to a number of factors such as the inability of the cells to produce sufficient amounts of antioxidants, nutritional deficiency of minerals or vitamins (Abd Ellah, 2010). When the production of ROS exceeds the capability of the body to detoxify these reactive intermediates, oxidative stress would develop (Mena et al., 2009). Oxidative stress can be induced by variety of factors such as radiation or exposure to heavy metals and xenobiotics (e.g carbon tetrachloride). This may lead to drastic harm to the body such as membrane damage, mutations due to attenuation of DNA molecules, and disruption to various enzymatic activities in metabolism of the body (McGrath et al., 2001; Valko et al., 2006; Chanda and Dave, 2009).
Medicinal plants are important sources of antioxidants (Rice, 2004). Antioxidants stabilize or deactivate free radicals, often before they attack targets in biological cells (Nunes et al., 2012). Natural antioxidants either in the form of raw extracts or their chemical constituents are very effective in preventing the destructive processes caused by oxidative stress (Zengin et al., 2011). Recently interest in naturally occurring antioxidants has considerably increased for use in food, cosmetic and pharmaceutical products, because they are multifaceted in their multitude and magnitude of activity and provide enormous scope in correcting imbalance (Djeridane et al., 2006; Wannes et al., 2010). The beneficial medicinal effects of plant materials typically results from the combinations of secondary products present in the plant (Wink, 1999). Phytochemical constituents of medicinal plants (e.g. polyphenols, carotenoids, flavonoids, phenolics, vitamins C and E), act as antioxidants by preventing damages to cell membrane due to cellular oxidative processes that may result in diseases (Omoregie and Osagie, 2011). They are found in all parts of plants such as leaves, fruits, seeds, roots and bark (Mathew and Abraham, 2006).
Antioxidants are broadly divided into enzymic antioxidants and non enzymic antioxidants. Enzymic antioxidants include the superoxide dismutases, glutathione peroxidase and catalase (Klaunig and Kamendulis, 2004). Non-enzymic antioxidants, which include vitamin E, vitamin C, β-carotene, reduced glutathione, and coenzyme Q function to quench reactive oxygen species (Clarkson and Thompson, 2000). Antioxidants have various mechanisms such as prevention of chain initiation, binding of transition metal ion catalysts, decomposition of peroxides, prevention of continued hydrogen abstraction and radical scavenging (Rao et al., 2004). Many chemicals damage mitochondria, an intracellular organelle that produces energy, its dysfunction release excessive amount of oxidants which in turn damage hepatic cells. Activation of some enzymes in the cytochrome P450 system, such as CYP2E1, also leads to oxidative stress (Jaeschke et al., 2002).
Carbon tetrachloride (CCl4) is a well known hepatotoxin used in diverse experimental models (Singh et al., 2008). In addition to hepatic problems, it causes dysfunction of the kidneys, lungs, testis, brain, and blood by generating free radicals (Ozturk et al., 2003; Khan et al., 2009). Carbon tetrachloride (CCl4) is rapidly transformed to trichloromethyl radical (CCl3*) and its derivative trichloromethyl peroxy radical (CCl3OO*), generated by cytochrome P450 of liver microsomes (Brent and Rumack, 1993). These free radicals react with membrane lipids leading to their peroxidation (Singh et al., 2008). Membrane disintegration of hepatocytes with subsequent release of membrane associated enzymes and necrosis are some of the consequences of CCl4 induced liver damage.
1.2 Statement of Research Problem
Hepatotoxicity is one of the very common ailments resulting into serious debilities ranging from severe metabolic disorders to even mortality (Anil et al., 2010). Liver injury due to chemicals or infectious agents may lead to progressive liver fibrosis and ultimately cirrhosis and liver failure (Anand, 1999). According to the report published by USFDA, more than 900 drugs, toxins, and herbs have been reported to cause liver injury, and drugs account for 20 – 40% of all instances of hepatic failure (Soni et al., 2011).
Liver ailments represent a major global health problem. Chronic liver cirrhosis and drug induced liver injury is the ninth leading cause of deaths in western and developing countries (Bharani Srinivasan et al., 2009; Saleem et al., 2010).
In Nigeria, as in other parts of sub-saharan Africa, the major causes of liver cirrhosis include infections particularly chronic hepatitis B virus (HBV) infection (Otu, 1987 and Cook, 1980) and hepatitis C virus infection (Bojuwoye, 1996). There is increasing evidence that free radicals and reactive oxygen species play a crucial role in various steps that initiate and regulate the progression of liver diseases independently of the original agent (Jemal et al., 2007).
1.3 Justification
Liver diseases remain one of the major threat to public health and a worldwide problem (Asha and Pushpangadan, 1998). The use of medicinal plants with high level of antioxidant constituents has been proposed as an effective therapeutic approach for hepatic damages (Govind, 2011).
Many antioxidants have been used to protect organs from the free radical challenges. Several researches are attempting to explore the possibility of using herbs containing antioxidants as organ curative agents. In view of severe undesirable side effects of synthetic agents, there is a growing focus to follow systematic research methodology and to evaluate scientific basis for the traditional herbal medicines that are claimed to possess hepatocurative activity.
Though the stem bark of Detarium microcarpum is used traditionally in Nigeria as a valuable remedy to treat liver diseases, no detailed pharmacological investigation have been done to that respect. This is why the present study is being undertaken to investigate it‟s hepato curative and antioxidant activity.
1.4 AIM AND OBJECTIVES
1.4.1 Aim
The aim of this study is to investigate the hepatoprotective and antioxidant effect of ethyl acetate and n-butanol fractions of Detarium microcarpum stem bark in CCl4-induced liver damage in rats.
1.4.2 Specific objectives
To identify and quantify levels of some phytochemicals present in the crude extract, ethyl-acetate and n-butanol fractions of Detarium microcarpum stem bark.
To carry out in-vitro antioxidant activity (such as DPPH, reducing power and H202 radical scavenging activity) on ethyl-acetate and n-butanol fractions of Detarium microcarpum stem bark and determine the lethal dose (LD50) of the fraction with the highest activity.
To determine the effects of the fraction with the highest activity on some biochemical parameters (such as ALT, AST, ALP, Bilirubin, Total protein, Albumin, Urea and Creatinine) in CCl4 induced liver damage in rats.
To determine the effect of the fraction with highest activity on some endogenous antioxidant enzyme levels and lipid peroxidation in the tissues of the animals induced with CCl4.
Null Hypothesis
Ethyl-acetate and n-butanol fractions of Detarium microcarpum stem bark do not have any hepato curative and antioxidant effect on CCl4 induced liver damage in rats.
REFERENCES
Abbot, D. and Andrews, R.S. (1970). An Introduction to chromatography 2nd ed. Longman press, London. pp. 72-78.
Abd Ellah, M. (2010). Involvement of free radicals in animal diseases. Comparative Clinical Pathology. 19(6):615–619.
Abreu, P. and Relva, A. (2002). Carbohydrates from Detarium microcarpum bark extract. Carbohydrate Research. 337: 1663-1666.
Abreu, P.M, Martins, E.S, Kayser, O, Bindseil, K.U, Siems, K, Seemann, A and Frevert J. (1999). Antimicrobial, antitumor and antileischmania screening of medicinal plants from Guinea-Bissau. Phytomedicine 6 (3):187-195.
Abreu, P.M, Rosa, V.S, Araujo, E, Canda, A.B. Kayser, O, Bindseil, K.U, Siems, K and Seemann, A, (1998). Phytochemical analysis and antimicrobial evolution of
Detarium microcarpum bark extracts. Pharmaceutical and Pharmacology Letters 8, 107-108.
Abulude, F.O., Onibon, V.O. and Oluwatoba, F. (2004). “Nutritional and nutritional compositions of some tree barks”. Nigerian Journal of Basic and Applied Sciences, 13: 43-49.
Aebi, H. (1984). “Catalase in Vitro” Methods in Enzymology 105, 121-126.
Ahsan, M.R, Islam, K.M and Bulbul, I.J. (2009). Hepatoprotective activity of Methanol Extract of some medicinal plants against carbon tetrachloride-induced hepatotoxicity in rats. European Journal of Scientific Research. 37(2): 302-310.
Akah, P.A, Nworu, C.S, Mbaoji, F.N, Nwabunike, I.A and Onyeto, C.A. (2012). Genus Detarium: Ethnomedicinal, phytochemical and pharmacological profile. Phytopharmacology, 3(2) 367-375
Alexander, R.R. and J.M. Grifiths, (1993). Haemoglobin Determination by the Cyanomethaemoglobin Method. In: Alexander, R.R. and J.M. Grifiths (Eds.), Basic Biochemical Methods. John Wiley and Sons, New York, pp: 188-189.
Amin, A and Mahmoud-Ghoneim, D. (2009). Zizyphus spina-christi protects against carbon tetrachloride-induced liver fibrosis in rats. Food and Chemical Toxicology., 47(8): 2111-2119.
Anand, B. (1999). Cirrhosis of liver. West L Med.171:110-115.
Anhwange, B.A, Ajibola, V.O and Oniye, S. (2004). Chemical studies of the seeds of
Moringa oleifera (Lam) and Detarium microcarpum (Guill and Sperr). Journal of Biological Science. 4(6): 711-715.
Anil, K.V, Kumar, K.V, Satish, R., Rama, T. Anil, K, Babul, D. and Samhitha, J. (2010). Hepatoprotective Effect of Flemingia Strobilifera R.Br. on Paracetamol induced Hepatotoxicity in Rats. International Journal of PharmTech Research. 2(3), 1924-1931.
Ansah, C, Dadzeasah, P.E and Asiamah, E. (2013). Aqueous stem bark extract of Spathodeacampanulata (p. Beauv) Modulates carbon tetrachloride Induced hepatic damage in rats. American Journal of Pharmacology and Toxicology 8 (1): 39-50.
Asha, V.V. and Pushpangadan, P. (1998). Preliminary evaluation of the anti-hepatotoxic activity of Phyllanthus kozhikodianus, Phyllanthus maderspatensis and Solanum indicum. Fitoterapia 59: 255-259.
ASTME, (1987). Standard Method for Estimating Acute Oral Toxicity in Rats, American Society for Testing and Materials, Philadelphia, 1163.
Augustyniak, A, Wazkilwicz E and Skrzydlewaka, E. (2005). Preventive action of green tea from changes in the liver antioxidant abilities of different aged rats intoxicated with ethanol. Nutrition, 21: 925-932.
Balamurugan, M, Parthasarathi, K, Cooper, E.L and Ranganathan, L.S (2009). Anti-inflammatory and anti-pyretic activities of earthworm extract-Lampito mauritii (Kinberg). Journal of Ethnopharmacology 121(2): 330-332.
Baranisrinivasan, P, Elumalai, E. K, Sivakumar, C, Therasa, S. V and David, E. (2009). Hepatoprotective effect of Enicostemma littorale blume and Eclipta alba during ethanol induced oxidative stress in albino rats. International Journal of Pharmacology. 5(4) 268–272.
Bartels, H. Bohnzer, M. and Heierli, C. (1972). Serum creatinine determination without protein precipitation. Clinical chemistry Acta, 37: 193-197.
Bataller, R and Brenner, D.A. (2005). Liver fibrosis. Journal of Clinical Investigation. 115:209–218.
Battu, G.R and Kumar, B.M. (2012). In-vitro antioxidant activity of leaf extract of Aerva lanata Linn. International Journal of Pharmaceutical Science, 2(4):74-78
Baynes, J.W.D and Marek H. (2005). Medical Biochemistry. Elsevier Mosby. pp 411-421
Bhattacharya, H, Lun, L and Gomez, R. (2005): Biochemical effects to toxicity of CCl4 on rosy barbs (Puntius conchonius). Our Nature 3:20–25.
Bhawna, S and Kumar, S.U (2010). Hepatoprotective activtity of some indigenous plants. International Journal of PharmTech Research. 2(1), 568-572
Bigoniya, P, Singh, C. S and Shukla, A. (2009). A Comprehensive Review of Different Liver Toxicants Used in Experimental Pharmacology. International Journal of Pharmaceutical Sciences and Drug Research; 1(3): 124-135
Bodakhe, S.H and Ram, A. (2007). Hepatoprotective Properties of Bauhinia variegata Bark Extract. Yakugaku Zasshi., 127: 1503-1507.
Boham BA and Kocipai-Abyazan R. (1974).“Flavonoids and condensed tannins from leaves of Hawaiian Vaccinium vaticulatum and V. calycinium” Pacific Science. 48: 458-463.
Boigk, G, Stroedter, L, Herbst, H, Waldschmidt, J, Riecken, E. O and Schuppan, D. (1997). Silymarin retards collagen accumulation in early and advanced biliary fibrosis secondary to complete bile duct obliteration in rats. Hepatology 26(3): 643-649.
Bojuwoye B.J (1996). Chronic diseases in Nigeria: An overview. Dokita. 23(1):1-4.
Borar, S, Punia, P and Kalia, A.N. (2011). Anitoxidant potential of n-butanol fraction from extract of Jaminum mesnyi Hance leaves. Indian Journal of Experimental Biology. 49: 39-43.
Brent, J and Rumack, B. (1993). Role of free radicals in toxic hepatic injury. II. Are free radicals the cause of toxin-induced liver injury? Journal of Toxicology:Clinical Toxicology. 31: 173-96.
Bruce, RD (1987). A confirmatory study of Up-and-Down Method of Acute Oral Toxicity Testing. Fundamental and Applied Toxicology. 8, 97 – 100.
Burk, R.F, Lane, J.M and Patal K. (2008). Relationship of oxygen and gluthathione in protection against CCl4 induced hepatic microsomal lipid peroxidation and covalent binding in the rat. Journal of clinical investigation, 74; 1996-2001.
Cavin, A.L, Hay, A.E, Marston, A, Stoeckli-Evans, H, Scopelliti, R, Diallo, D and Hostettman, K (2006). Bioactive Diterpenes from the Fruits of Detarium microcarpum. Journal of Natural Products. 69(5): 768-773.
Chan, E.W.C, Lim, Y.Y and Omar, M. (2007). Antioxidant and antibacterial activity of leaves of Etlingera species (Zingiberaceae) in Peninsular Malaysia. Food Chemistry, 104 (4): 1586-1593.
Chanda, S. and Dave, R. (2009). “In vitro models for antioxidant activity evaluation and some medicinal plants possessing antioxidant properties: an overview,” African Journal of Microbiology Research, 3(13) 981–996.
Chun, O.K, Kim, D.O. and Lee, CY. (2003). Superoxide radical scavenging activity of the major polyphenols in fresh plums. Journal of Agriculture and Food Chemistry 51: 8067-8072.
Clarkson, P.M and Thompson, H.S. (2000) Antioxidants: what role do they play in physical activity and health? American Journal of Clinical Nutrition 72 66. (S): 637-46
Contu S. (2012). Detarium microcarpum. IUCN Red List of Threatened Species. (2).
Cook, G.C. (1980). Liver diseases in Tropical Gastroenterology. Oxford University Press, New York Delhi, pp. 95 – 117.
Cotran, R.S, Kumar, V and Robbins, S.L. (1994). Genetic Disorders. In: Cotran RS, Kumar V, Robbins SL. Pathologic Basis of Disease. Edn 5, W B Saunders Co, Philadelphia, pp143.
Crocenzi, F.A and Roma, M.G. (2006). Silymarin as a new hepatoprotective agent in experimental cholestasis: new possibilities for an ancient medication. Current Medicinal Chemistry 13(9): 1055-1074.
Cui, C.P, Wei, P, Liu, Y, Zhang, D.J, Wang, L.S and Wu, C.T (2009). The protective role of Hematopoietin Cn on liver injury induced by carbon tetrachloride in rats. Hepatology Research., 39(2): 200-206
Cullen, J.H. (2005). Mechanistic classification of liver injury. Toxicologic Pathology 33: 6-8.
Dalziel, J.M. 1955. The Useful Plants of West Africa. Crown for Oversea Government and Admin. 4 Mill Bank London, pp: 186-188.
Dehmlow, C.; Erhard, J. and de Groot, H. (1996). Inhibition of Kupffer cell functions as an explanation for the hepatoprotective properties of silibinin. Hepatology, 23(4) 749-754.
Djeridane, A, Yousfi, M, Nadjemi, B, Boutassouna, D, Stocker, P and Vidal, N. (2006). Antioxidant activity of some Algerian medicinal plants extracts containing phenolic compounds. Food Chemistry. 97:654–660.
Doumas, B.T, Watson, W and Homer G. (1997). Albumin standards and the measurement of serum albumin with bromcresol green. Clinica Chimica Acta. 258(1) : 21-30.
Duncan, D.B. (1955). Multiple range and multiple F-test. Biometrics, 11, 1–42.
Ebi, G.C and Afieroho, O.E. (2011). Phytochemical and antimicrobial studies on Detarium microrcarpum Guill and Perr (Caesalpiniaceae) seeds coats. African Journal of Biotechnology 11(3), 457-462.
Ejizu, C.I. (1986). Ofo: Igbo ritual symbols. Fourth Dimension publishers, Enugu, Nigeria pp 190.
El- Shafey, A.A.M, Seliem, M.M.E, El-Mahrouky, F, Gabr, W.M, and Kandil R.A. (2011). Some physiological and biochemical effects of Oshar extract and abamectin biocide on male albino rats. Journal of American Science. 7(12): 254-261
El-Olemy, M.M, Al-Muhtadi, F.J and Afifi, A.F.A. (1994). Experimental phytochemistry: A laboratory manual. King Saud University Press, Saudi Arabia, 21-27.
Encyclopaedia Britannica, (2003) Inc.; www.britannica.com
Etuk, E.U, Agaie, B.M, Ladan, M.J and Garba, I. (2009). The modulatory effect of Cochlospermum tinctorium a rich aqueous root extract on liver damage induced by carbon tetrachloride in rats. African Journal of Pharmacy and Pharmacology. 3(4). 151-157.
Famhy, S.R and Hamdi, S.A.H. (2011). Curative effect of the egyptian marine Erugosquilla massavensis extract on carbon tetrachloride-induced oxidative stress in rat liver and erythrocytes. European Review for Medical and Pharmacological Sciences. 15: 303-312.
Fawcett, J.K and Scott, J.E. (1960): A rapid and precise method for the determination of urea. Journal of Clinical Pathology. 13:156-159.
Field, K. M, Dow, C and Michael, M. (2008). Liver function in oncology: biochemistry and beyond. The Lancet Oncology. 9 (11) 1092–1101.
Fraga, C. G., Leibovitz, B. E. and Tappel, A. L. (1988). Lipid peroxidation measured as thiobarbituric-reactive substances in tissue slices: characterization and comparison with homogenates and microsomes. Free Radical Biology and Medicine. 4:155-161.
Fraschini, F. Demartini, G. and Esposti, D. (2002). Pharmacology of silymarin. Clinical Drug Investigation, 22(1) 51–65.
Friedman, LS, Martin, P, Munoz, S.J. (1996). Liver Function Tests and the Objective Evaluation of the Patient with Liver disease. In: Zakin D, Boyer TD. Hepatology: A Textbook of Liver Disease. Edn 3, W B Saunders Co, Philadelphia, pp 791-833.
Fromenty, B. and Pessayre, D. (1995). Inhibition of mitochondrial beta-oxidation as a mechanism of hepatotoxicity. Pharmacology and Therapeutics 67:101–154.
Garba, S, Andrew, K and Gayus, J. (2013). Antibacterial activities of total alkaloids extracted from some medicinal plants. Topclass Journal of Herbal Medicine. 2(9) 217-222.
Govind, P. (2011). Medicinal plants against liver diseases. Internatioal Journal of Pharmaceutical Research. 2:115–121.
Gülçin, I. Alici H.A. and Cesur, M, (2005). Determination of in-vitro antioxidant radical scavenging activities of propofol. Chemical Pharmarceutical Bulletin. 53:281– 285.
Gupta S.K (2004). Drug screening methods. First ed. New Delhi: jaypee Brothers, medical Publishers; pp 463-64.
Gutierrez-Ruiz, M.C, Gomez-Quiroz, L.E. (2007). Liver fibrosis: searching for cell model answers. Liver International 27:434–439.
Halliwell, B. (1995). How to characterize an antioxidant: an update. Biochemical Society Symposia. 61: 73-101.
Harborne, J.B (1973). Phytochemical Methods: A Guide to modern Techniques of Plant analysis. Chap,amA. And Hall London, pp 279.
Harsh, M. (2005). Textbook of Pathology. Edn 5, Jaypee Brothers: Medical Publishers
(P) Ltd., New Delhi, pp 22-24 and 608-668.
Iful, S. (2008). Studies on the antivenorm activities of the aqueous extracts of Paullinia pinnata and Detarium microcarpum. Ph.D. Thesu University of Jos, Nigeria.
Iniaghe, O.M., Malomo, S.O and Adebayo, J.O. (2008). Journal of Medicinal Plants Research. 2 (10) 301-305.
Irvine, F.R. (1961). The Woody Plants of Ghana with Special References to Their Uses. Oxford University Press London. pp 863.
Jaeschke, H, Gores, G.J, Cederbaum, A.I, Hinson, J.A, Pessayre, D and Lemasters, J.J. (2002). Mechanisms of hepatotoxicity. Toxicological Science; 65: 166-176.
Jan, S, Khan, M.R, Rashid, U and Bokhari, J. (2013). Assessment of Antioxidant Potential, Total Phenolics and Flavonoids of Different Solvent Fractions of Monotheca Buxifolia Fruit. Public Health Research Perspective. 4(5): 246–254.
Jemal, A, Siegel R and Ward, E. (2007). Cancer statistics. Cancer Journal for Clinicians. 57:43–66.
Jendrassik, L. and Grof, P. (1938). Colorimetric Method of Determination of bilirubin. Biochemische Zeitschrift. 297:81-82
Jensen, J.E, Steinberg, S.E, Freese, P and Marino, E. (2004). Liver function tests. Journal of Digestive Disorder. 6: 1-3.
Johnson, D.E. and Kroening, C. (1998). Mechanism of carbon tetrachloride toxicity in cultured rat hepatocytes. Pharmacology and Toxicology., 83: 231-239.
Junnila, M, Rakho, A, Sukura, A and Lindberg, L.A. (2000). Reduction of carbon tetrachloride induced hepatotoxic effects by oral administration of betaine in male hans-wistar rats: a morphometric histological study. Veterinary Pathology. 37: 231-238
Kaboré, C. (2005). Aménagement des forêts au Sahel–Point sur vingt années de pratiques au Burkina Faso. Ouagadougou, Burkina Faso. Ministère de l’Environnement et de l’Eau.
Kalava, S.V. and Menon, S.G. (2012). Protective efficacy of the extract of volvariella volvacea (bulliard ex fries) singer. against carbon tetrachloride induced hepatic injury. International Journal of Pharmaceutical Sciences and Research, Vol. 3(8): 2849-2856.
Kaplan, M.M. (1993). Laboratory tests. In: Schiff L, Schiff ER. Diseases of the Liver. Edn 7, J B Lippincott, Philadelphia, pp 108-144.
Kaplowitz, N.T.Y, Simon, F.R and Stolz, A. (1986). Drug induced hepatotoxicity. Annals of Internal Medicine. 104: 826-839. 105
Keay, R.W.J, Onochie C.F.A, and Standfield, D.P (1964). Nigeria Trees vol. 1, Dept. of Forest Research, Ibadan Nigeria.
Khan, M.R and Siddique, F. (2012). Antioxidant effects of Citharexylum spinosum in CCl4 induced nephrotoxicity in rat. Experimental and Toxicologic Pathology. 64:349-55.
Khan R.A, Khan M.R and Sahreen S. (2012). CCl4–induced hepatotoxicity: Protective effect of rutin on p53, CYP2E1 and the antioxidantive status in rats. BMC Complementary and Alternative Medicine, 12:178.
Khan, M.R., Rizvi, W., Khan, G.N., Khan, R.A and Shaheen, S. (2009): Carbon tetrachloride-induced nephrotoxicity in rats: protective role of Digera muricata. Journal of Ethnopharmacology. 122: 91-99.
Khanum, R, Mazhar, F and Jahangir, M. (2015). Antioxidant evaluations of polar and non-polar fractions of Cajanus cajan seeds. Journal of Medical Plants and Research. 9(6) 193-198.
Kim, H.Y, Kim, J.K, Choi, J.H, Jung, J.Y, Oh, W.Y, Kim, D.C and Lee, S.M (2010). Hepatoprotective effect of pinoresinol on carbon tetrachloride-induced hepatic damage in mice. Journal of Pharmacology and Science., 112(1):105-112.
King, E.J and Amstrong, A.R, (1980). Method of King and Amstrong In; Varley, H, Gowenlock, AH, Bell, M. Practical Clinical Biochemisty, 5th Edn., William Heinmann Medical Books Ltd, London. Pp 897.
Klaunig, J.E, Kamendulis, L.M (2004). The role of oxidative stress in carcinogenesis. Annual Review of Pharmacology and Toxicology. 44:239-267.
Kmiec, Z. (2001) Cooperation of liver cells in health and disease. Advances in Anatomy, Embryology and Cell Biology, 161, III-XIII, 1-151.
Kouyaté, A.M and Lamien, N. (2011). “Detarium microcarpum, sweet detar,” Conservation and Sustainable Use of Genetic Resources of Priority Food Tree Species in sub-Saharan Africa. Biodiversity International 4: 4.
Kouyaté A.M and van Damme P. (2006). “Medicinal plants/Plantes médicinales: Detarium microcarpum Guill. & Perr.” Prota 11 (1).
Kouyate, AM (2005). Aspects ethnobotaniques et etude de la variability morphologique, biochimique et phenologique de Detarium microcapum. Guill and perr. Au Mali. PhD thesis faculty of Agriculture and Applied Biological Sciences. University of Gent. Gent. Belguim pp. 207.
Lewis, J.H. (2000). Drug-induced liver disease. Medical Clinics North America 84:1275–1311.
Li ,X, Wu, X and Huang, L. (2009). Correlation between antioxidant activities and phenolic contents of Radix Angelicae sinensis (Danggui). Molecule 14, 5349-5361.
Lim, H.K, Kim, H.S, Choi, H.S, Oh, S, Jang, C.G and Choi, J. (2000). Effects of acetyl bergenin against D-galactosamine induced hepatotoxicity in rats. Pharmacological Research 42:471.
Liston, H.L, Markowitz, J.S and Devane, C.L. (2001). Drug glucuronidation in clinical psychopharmacology. Journal of Clinical Psychopharmacology 21(5): 500-515.
Liu, J, Tan, H, Sun, Y, Zhou, S, Cao, J and Wang F (2009). The preventive effects of heparin-superoxide dismutase on carbon tetrachloride-induced acute liver failure and hepatic fibrosis in mice. Molecular and Cellular Biochemistry., 327(1-2): 219-228.
Lorke, D. (1983). A new approach to practical acute toxicity testing. Archives of Toxicology. 54, 275
Lowry, O.H, Rosebrough, N.J, Farr, A.L and Randall, R.J. (1951). Protein measurement with the Folinphenol reagent. Journal of Biology and Chemistry., 193, 265–267.
Mahmood, N, Pizza, C, Aquino, P, de Tommasi, N, Piacente, S, Colman, S, Buma, A and Itay, A.J. (1993). Inhibition of HIV infection by flavonoids. Antiviral Research 22, 189-199.
Manoj, B. and Aqued, K. (2003). Protective effect of Lawson Alba L. against CCl4-induced hepatic damage in albino rats. Indian Journal of Experimental Biology. 4:85-87.
Marina, N. (2006). Hepatotoxicity of antiretrovirals: incidence, mechanisms and management. Journal of Hepatology. 44 (1) S132–S139.
Martin, J.P, Dailey, M and Sugarman, E. (1987). Negative and positive assays of superoxide dismutase based on hematoxylin autooxidation. Archives of Biochemistry and Biophysics 255 329–336.
Mathew, S and Abraham, T. (2006). In-vitro antioxidant activity and scavenging effects of Cinnamomum verum leaf extract assayed by different methodologies. Food and Chemical Toxicology 44: 198-206.
Mazumder, U.K, Gupta, M, Chakraborty, G and Monda, S.K. (2006). In-vitro antioxidant activity of Diospyros mamabarica Kostel bark. Indian Journal of Experimemtal Biology. (44) 39-44.
McGrath, L.T, McGleenon, B.M, Brennan, S. McColl, D, McIlroy, S. and Passmore, A.P (2001). “Increased oxidative stress in Alzheimer’s disease as assessed with 4-hydroxynonenal but not malondialdehyde,” Quarterly Journal of Medicine. 94(9) 485–490.
Medina, J and Moreno-Otero, R. (2005). Pathophysiological basis for antioxidant therapy in chronic liver disease. Drugs, 65(17):2445-2461.
Mena, S. Ortega, A and Estrela, J.M. (2009). “Oxidative stress in environmental-induced carcinogenesis,” Mutation Research, 674(1-2) 36–44.
Mir, M.A, Sawhney, S.S and Jassal, M.M.S (2013). Qualitative and quantitative analysis of phytochemicals of Taraxacum officinale. Wudpecker Journal of Pharmacy and Pharmacology. 2(1) 001 – 005.
Mojzisova, G and Kuchta, M. (2001). Dietary flavonoids and risk of coronary heart disease. Physiological Research. 50: 529-535.
Muller, D, Enderle, G.J, Low, O, Dietze, E and Krell, H. (1996). Bile ductular proliferation and altered leukotriene elimination in thioacetamide-induced fibrosis of rat liver. Journal of Hepatology 25:547–553.
Navarro, V.J and Senior, J.R. (2006). Drug-related hepatotoxicity. New England Journal of Medicine. 354:731-9.
Nazarian, R. M., Van Cott, E. M., Zembowicz, A and Duncan, L.M. (2009). Warfarin-induced skin necrosis. Journal of the American Academy of Dermatology 61, 325-332.
Nebert, D.W and Dalton, T.P (2006). The role of cytochrome P450 enzymes in endogenous signalling pathways and environmental carcinogenesis. Nature Reviews Cancer, 6, 947 60.
Nebert, D.W and Gonzalez, F.J. (1987). P450 genes: structure, evolution, and regulation. Annual Review of Biochemistry, 56, 945-93.
Njoku, O.U, Obioma, U and Franck, E.U (1999). Investigation on some nutritional and toxicological properties of Afzelia africana and Detarium microcarpum seed oil. Bolle Chimico Farmaceutico 138, 165-168.
Nunes, P.X, Silva, S.F, Guedes, R.J and Almeida S. (2012). Biological oxidations and antioxidant activity of natural products, Phytochemicals as nutraceuticals – Global Approaches to Their Role in Nutrition and Health. Croatia: Tech publishers. pp 1-20.
Obdoni, B.O and Ochuko, P.O. (2001). “Phytochemical studies and comparative efficacy of the crude extracts of some Homostatic plants in Edo and Delta States of Nigeria” Global Journal of Pure and Applied Science. 8: 203-208
Obun, C.O. Yahaya, S.M, Kibon, A.A. Olafadehan, O.A. and Alison, S.D. (2010). Evaluation of Detarium microcarpum pulp meal as food ingredient in rabbits diets. Electronic Journal of Environmental, Agriculture and Food Chemistry, 9 (2) 308 – 314.
Ogasawara, J, Watanabe-Fukunaga, R and Adachi, M. (1993). Lethal effect of the anti-Fas antibody in mice. Nature 364:806–809.
Ogeturk, M, Kus, I, Colakoglu, N, Zararsiz, I, Ilhan, N and Sarsilmaz, M. (2005): Caffeic acid phenethyl ester protects kidneys against carbon tetrachloride toxicity in rats. Journal of Ethnopharmacology. 97,273-280.
Ogeturk, M, Kus, I, Kavakli, A, Zararsiz, I, Ilhan, N and Sarsilmaz, M. (2004). Effect of melatonin on carbontetrachloride induced changes in rat serum. Journal of Physiology and Biochemistry. 60 (3) 205–210.
Okolo, C.E, Akah, P.A and Uzodinma, S.U. (2012). Antidiabetic activity of root extract of Detarium microcarpum. (Fabicase) Guill & Perr. Phytopharmacology 3 (1), 12-18.
Okorie, O, Okonkwo, T.J.N, Nwachukwu, N. and Okeke I. (2010). Potentials of Detarium microcarpum (guill and Sperr) seed oil as a matrix for the formulation of haloperidol injection. 5(1).
Okorie, O. (2004). Some Physicochemical and in vitro Bioadhesive Properties of Defatted Detarium microcarpum Guill and Sperr (Family:Caesalpiniaceae) Gum. Ph. D. Thesis (Unpublished), Department of Pharmaceutical Technology and Industrial Pharmacy, University of Nigeria, Nsukka, Enugu State, Nigeria.
Olajire, A. A and Azeez, L (2011). Total antioxidant activity, phenolic, flavonoid and ascorbic acid contents of Nigerian vegetables. African Journal of Food Science and Technology. 2(2) 022-029.
Olaleye, M, Afolabi, C, Adebayo, A, Ogunboye, A and Akindahunsi A, (2010). Antioxidant activity and hepatoprotective property of leaf extracts of Boerhaavia diffusa Linn against acetaminophen-induced liver damage in rats, Food and Chemical Toxicology. 48: 2200-2205
Olugbuyiro, J.A.O and Akinbohun, O.F (2012). In-vitro activity of Bryophyllum pinnatum and Detarium microcarpum plants against Mycobacterium tuberculosis and other bacteria. Natural Products Research Bulletin 1: 012-018.
Olugbuyi, J.A.O, Moody, J.O and Hamann, M.T. (2012). In-vitro activity of Bryophyllum pinnatum and Detarium microcarpum plants against Mycobacterium tuberculosis and other bacteria. Natural Products Research Bulletin 1: 012-018.
Olugbuyi J.A.O, Moody, J.O and Hamana, M.T. (2009). Inhibitory activity of Detarium microcarpum extract against hepatitis C virus. African Journal of Biomedical Research 12(2), 149- 151.
Omoregie, E.S. and Osagie, A.U. (2011). Effect of Jatropha tanjorensis leaves supplement on the activities of some antioxidant enzymes, vitamins and lipid peroxidation in rats. Journal of Food Biochemistry, 35(2): 409-424.
Orisakwe, O.E, Obi, E, Udemezue, O.O and Orish, V.N. (2003) Effect of halofantrine on testicular architecture and testosterone level in guinea pigs. European Bulletin of Drug Research., 11(4) 105-109.
Otu, A.A (1987). Hepatocellular carcinoma, hepatic cirrhosis and hepatitis B virus infection in Nigeria. Cancer. 60(10): 2581 – 2585
Oyaizu, M., (1986). Studies on products of browning reaction: Antioxidantive activity of products of browning reaction prepared from glucosamine. Japanese Journal of Nutrition, 44: 307-315.
Ozturk, F, Ucar, M, Ozturk, I.C, Vardi, N and Batcioglu, K. (2003). Carbon tetrachloride-induced nephrotoxicity and protective effect of betaine in Sprague-Dawley rats. Urology, 62(2): 353-356.
Palani, S, Raja, S, Venkadesan, D, Karthi, S, Sakthivel, K and Senthil, K.B. (2009). Antioxidant activity and hepatoprotective potential of Terminalia pallida. Archives of Applied Science and Research. 1 (1) 18–28.
Pari, L and Kumar, N.A. (2002). Hepatoprotective activity of Moringa oleifera on antitubercular drug-induced liver damage in rats. Journal of Medical Foods. 5: 171-177.
Parr, S. and Bolwell, G (2000). Phenols in the plant and in man. The potential for possible nutritional enhancement of the diet by modifying the phenols content or profile. Journal of Science of Food and Agriculture., 80: 985-1012.
Patel, A, Patel, A, Patel, A, Patel, N.M. (2010). Estimation of Flavonoid, Polyphenolic Content and in-vitro Antioxidant Capacity of leaves of Tephrosia purpurea Linn. (Leguminosae). International Journal of Pharmaceutical Science and Research. 1(1) 66-77.
Patel, T, Roberts, L.R, Jones, B.A and Gores, G.J. (1998). Dysregulation of apoptosis as a mechanism of liver disease: an overview. Seminars in Liver Disease 18(2): 105-114.
Pessayre, D, Berson, A, Fromenty, B and Mansouri, A. (2001). Mitochondria in steatohepatitis. Seminar in Liver Disease. 21:57–69.
Post-White, J, Ladas, E.J and Kelly, K.M. (2007). Advances in the use of milk thistle (Silybum marianum). Integrative Cancer Therapies 6(2): 104-109.
Proskuryakov, S.Y, Konoplyannikov, A.G and Gabai, V.L. (2003). Necrosis: a specific form of programmed cell death? Experimental Cell Research 283, 1-16.
Raffray, M and Gerald M.C. (1997). Apoptosis and necrosis in toxicology: A continuum or distinct modes of cell death? Pharmacology & Therapeutics 75, 153-177.
Rao, C.V, Ojha, S.K, Radhakrishnan, K, Govindrajan, R, Rastogi, S, Mehrotra, S and Pushpangadan, P. (2004). Antiulcer activity of Utleria saclicifolia rhizome extract. Journal of Ethnopharmacology. 91: 243-249.
Reitman, S and Frankel A. (1957). Colorimetric method for determination of serum glutamate oxaloaectate and glutamic pyruvate transaminase. American Journal of Clinical Pathology. 28: 56-58.
Riaz, T, Abbasi, M.A, Aziz, U.R Shahzadi, T, Ajaib, M and Khan K.M. (2012). Phytochemical screening, free radical scavenging, antioxidant activity and phenolic content of Dodonaea viscose Jacq. Journal of Serberia Chemical Society 77 (4) 423–435
Rice, E. (2004). Flavonoids and isoflavones: absorption, metabolism and bioactivity. Free Radical Biology and Medicine. 36: 827-828.
Rodriguez-Antona, C and Ingelman-Sundberg, M. (2006). Cytochrome P450 pharmacogenetics and cancer. Oncogene 25, 1679-91.
Ross, A.W and Wilson, K.J.W (2005). Anatomy and Physiology in Health and illness, Ninth Edition, published by Elsevier Ltd., 307-310, 333-336.
Saleem, T, Chetty, C, Ramkanth, S, Rajan, V, Kumar, K and Gauthaman, K. (2010). Hepatoprotective Herbs–A Review. International Journal of Research in Pharmaceutical Science. 1:1-5.
Saller, R. Melzer, J. Reichling, J. Brignoli, R. and Meier, R. (2007). An updated systematic review of the pharmacology of silymarin. Forschende Komplementmedizin. 14(2) pp. 70- 80.
Samir, S. (2001). Hepatotprotective Natural Products. Journal of Advance Pharmaceutica Research and Bioscience. 2(5), 110-111
Savadogo, P.., Tigabu, M.., Sawadogo, L and Odén, P.C. (2007). Woody species composition, structure and diversity of vegetation patches of a Sudanian savanna in Burkina Faso. Bois et Forêts des Tropiques 294(4): 5–20.
Şener, G, Şehirli, A.Ö and Ayanoğlu-Dülger, G. (2003). Protective effects of melatonin, vitamin E, and N-acetylcysteine against acetaminophen toxicity in mice: a comparative study. Journal of Pineal Research. (35) 61-8
Shah, N.A, Khan, M.R, Sattar, S, Ahmad, B and Mirza, B. (2014). HPLC-DAD analysis, antioxidant potential andanti-urease activity of Asparagus gracilis collected from District Islamabad. BMC Complementary and Alternative Medicine, 14:347.
Sharma, M.P and Ahuja, V. (1997). Amoebic liver abscess: Clinicians perspective. Bombay Hospital Journal 39, 615-619.
Sherlock, S.D. and James, D. (2002) Diseases of the liver and biliary system. Blackwell Publishing. pp 26-34.
Singh, N, Kamath, V, Narasimhamurthy, K and Rajini, PS. (2008). Protective effects of potato peel extract against carbon tetrachloride-induced liver injury in rats. Journal of Environmental Toxicology and Pharmacology. 6:242–246.
Singh, N and Rajini, P.S. (2004). Free radical scavenging activity of an aqueous extract of potato peel. Food Chemistry 85, 611-616.
Smith, G, Stubbins, M.J, Harries, L.W and Wolf, C.R. (1998). Molecular genetics of the human cytochrome P450 monooxygenase superfamily. Xenobiotica 28: 1129-1165.
Sofowora, A (1993). Medicinal Plants and Traditional Medicine in Africa. Chichester John Wiley and sons New York, pp. 97-145.
Soni, M, Mohanty, P.K and Jaliwala, Y.A. (2011). Hepatoprotective activity of fruits of Prunus Domestica. International Journal of Pharma And Bioscience; 2(2): 439-453.
Sreelatha, S, Padma, R and Umadevi, M. (2009). “Protective Effects of Coriandrum sativum Extracts on Carbontetrachloride induced Hepatotocity in Rats,” Food and Chemical Toxicology. (47) 702-708.
Srivastava, A and T. Shivanandappa, (2010). “Hepatoprotective Effect of the Root
Extract of Decalepishamiltonii against Carbon Tetrachloride-Induced Oxidative Stress in Rats,” Food Chemistry, 118 (2) 411-417.
Sunil, M, Kunduri, R.D, Sashi, B.B and Jyotirmoyee J. (2012). Protective and curative effect of poly herbal formulation containing indigenous medicinal plants against various hepatotoxic agents in rats. Asian Pacific Journal of Tropical Biomedicine (2012) S1377 – S1381.
Szymonik-Lesiuk, S, Czechowska, G, Stryjecka-Zimmer, M, Słomka, M, Madro, A, Celiński, K and Wielosz M (2003). Catalase, superoxide dismutase, and glutathione peroxidase activities in various rat tissues after carbon tetrachloride intoxication. Journal of Hepatobiliary Pancreatic Surgery., 10(4): 309-315.
Tatiya, A.U, Surana, S.J, Sutar, M.Pand Gamit, N.H. (2012). Hepatoprotective effect of poly herbal formulation against various hepatotoxic agents in rats. Pharmacognosy Research 4:50-56.
Teselkin, Y.O, Babenkova, I.V, Kolhir, V.K, Baginskaya, A.I, Tjukavkina, N.A, Kolesnik, Y.A and Eichholz, A.A. (2000). Dihydroquercetin as a means of antioxidative defence in rats with tetrachloromethane hepatitis. Phytotherapy Research. 14 (3): 160.
Tortora, G.J and Grabowski, S.R (2003). Principle of Anatomy and Physiology. (10), 875- 879.
Tosun, M, Ercisli, S, Sengul, M, Oezr, H, Polat, T and Ozturk, E. (2009). Antioxidant properties and total phenolic content of salvia species from Turkey. Biological Research. 42, 175-181.
Trease, G.E, Evans, and W.C. (1989). A Textbook of Pharmacognosy. 13th edn. Bailliere Tindall Ltd.: London. pp 60-75.
Ujah, O. F, Ujah, I. R, Johnson, J.T and Oka, V.O. (2013). Effect of ethanolic leaf extract of Moringa olifera leaf on haematological and biochemical parameters of wistar rats. Journal of Natural Product and Plant Resources. 3 (2):10-14.
Usha, K., Mary, K.G and Hemalatha, P. (2008). Hepatoprotective effect of Hygrophila spinosa and Cassia occidentalis on carbon tetrachloride induced liver damage in experimental rats. Indian Journal of Clinical Biochemistry, 22(2): 132-135.
Valko, M. Rhodes, C.J, Moncol, J, Izakovic, M and Mazur M. (2006). “Free radicals, metals and antioxidants in oxidative stress-induced cancer,” Chemico-Biological Interactions, 160 (1) 1–40,
Valko, M. Leibfritz, D. Moncol, J. Cronin, M.T.D, Mazur, M. and Telser, J. (2007).
“Free radicals and antioxidants in normal physiological functions and human disease,” International Journal of Biochemistry and Cell Biology, 39 (1) 44–84.
Van-Burden, T and Robinson, W. (1981). Formation of complexes between protein and Tannin acid. Journal of Agriculture and Food Chemistry. 1: 77.
Vautier, H, Sanon, M and Moctar, S, (2007). SEED LEAFLET No. 122. Forest & Landscape Denmark. Hørsholm Kongevej 11. DK-2970 Hørsholm. (www.SL.ku.dk). Accessed August 26, 2010, at 15:37.
Vinson, J.A and Zhang, J. (2005). Black and Green tea equally inhibit diabetic cataracts in a Streptozotocin-induced rat model of Diabetes. Journal of Agriculture and Food Chemistry. 53, 3710 – 371.
Wallace, A.H. (2007). Principles and Methods of Toxicology. Healthcare USA, Inc., New York, 5th ed., pp. 369-453.
Wannes, W.A, Mhamdi, B, Sriti, J, Jemia, M.B, Ouchikh, O, Hamdaoui, G, Kchouk, M.E and Marzouk, B. (2010). Antioxidant activities of the essential oil and methanol extracts from myrtle (Myrtus communis var. italica L.) leaf, stem and flower. Food and Chemistry Toxicology, 48:1362–1370.
Waugh, A, Grant, A, Ross, A.W and Wilson, K.J.W (2001). Anatomy and Physiology in Health and Illness. Edn 9, Churchill Livingstone, Spain, pp 307-311.
Werck-Reichhart, D and Feyereisen, R. (2000). Cytochromes P450: a success story. Genome Biology. 1(6): 3003.1-3003.9
WHO (2001). Legal Status of Traditional Medicine and Complementary/ Alternative medicine: A world wide review. WHO Publishing 1, Geneva.
Williams, R.J, Spencer J.P and Rice-Evans, C. (2004). Flavonoids: Antioxidants or signalling molecules? Free Radical Biology and Medicine., 36: 838-849.
Wink, M. (1999). Introduction to Biochemistry, role and biotechnology of secondary products. In: M Wink, Ed, and Biochemistry of secondary product Metabolism. CRC Press, Boca Ratom, FL;PP. 1-16
www.westafricanplants.senckenberg.de/images/pictures/detarium_microcarpum_ms_21 51_552_ff9925.jpg. Accessed May 18, 2015, at 6:30.
Yousef, M.I, Awad, I.T and Mohamed, E.H. (2006). Deltamethrin-induced oxidative damage and biochemical alterations in rat and its attenuation by Vitamin E. Toxicology, 227(3):240-247.
Zatloukalova, J. (2008). Deregulation of cell proliferation and apoptosis by xenobiotics and cytostatics (Unpublished P.hD thesis). Masarykova Univerzita, Czech Republic.
Zayachkivska, O.S, Konturek, S.G, Drozdowicz, D, Konturek, P.C, Brzozowski, T and Ghegotsky, M.R. (2005). Gastroprotective effects of flavonoids in plant extracts.
Journal of Physiology and Pharmacology., 56: 219-231.
Zengin, G, Cakmak, Y.S, Guler, G.O and Aktumsek, A. (2011). Antioxidant properties of methanolic extract and fatty acid composition of Centaurea urvillei DC. subsp. hayekiana Wagenitz. Records of Natural Products , 5:123–132.
Zhao, J. Lahiri-Chatterjee, M, Sharma, Y and Agarwal, R. (2000). Inhibitory effect of a flavonoid antioxidant silymarin on benzoyl peroxide-induced tumor promotion, oxidative stress and inflammatory responses in SENCAR mouse skin.
Carcinogenesis 21(4): 811-816.
Zimmerman, H.J. (1999). Hepatotoxicity: The Adverse Effect of Drugs and other Chemicals on the Liver. 2nd Ed. Philadelphia: Lippincott, Williams & Wilkins. pp 427-456.
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