JBM  Vol.5 No.6 , June 2017
Biochemical and Haematological Effects of the Leaf Extract of Newbouldia laevis in Alloxan-Induced Diabetic Rats
Abstract: Dichloromethane-methanol (1:1) extract (DME) of N. laevis leaves was prepared by cold maceration. The effects of the extract on the haematological and some biochemical parameters of alloxan-induced diabetic rats were investigated. The results showed that the oral administration of the extract (250, 500, 1000 mg/kg) caused a significant (P < 0.5) and dose-dependent increase in red blood cell count (RBC) and its indices, as well as a significant (p < 0.05) and dose-dependent reduction in the platelet count and the white blood cells (WBC). The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were significantly (p < 0.05) decreased. This effect was not dose related. The serum levels of total bilirubin, urea and creatinine were significantly (p < 0.05) decreased. The serum total protein and total antioxidant status (TAS) significantly (p < 0.05) increased dose dependently. Overall, administration of DME has significant ameliorative effect on alloxan-induced anaemia and other haematological alterations in diabetes and this may be of immense benefits in the management of diabetes and its associated haematological complications. Improved liver and kidney functions as well as improved antioxidant status are beneficial in the management of chronic diseases such as diabetes.
Cite this paper: Osigwe, C. , Akah, P. and Nworu, C. (2017) Biochemical and Haematological Effects of the Leaf Extract of Newbouldia laevis in Alloxan-Induced Diabetic Rats. Journal of Biosciences and Medicines, 5, 18-36. doi: 10.4236/jbm.2017.56003.

[1]   Ogbe, F.M.D., Eruogunn, O.L. and Wagboe, M. (2009) Plants Used for Female Reproductive Healthcare in Oredo Local Government Area, Nigeria. Scientific Research and Essays, 4, 120-130.

[2]   Hutchison, J. and Dalziel, J.M. (1963) Flora of West Tropical Africa. Vol. II, Crown Agents for Overseas Government and Administration, London, 435-436.

[3]   Oliver-Bever, B. (1986) Medicinal plants in Tropical West Africa. Cambridge University Press, Cambridge, 117-168.

[4]   Burkill, H.M. (1997) The useful plants of West Tropical Africa. 2nd Edition, Vol. 4 (Families M-R), Royal Botanic Gardens, Kew.

[5]   Iwu, M.M. (2000) Handbook of African Medicinal Plants. CRC Press, Inc., London, 19.

[6]   Ogunlesi, M., Okiei, W., Ofor, E. and Awonuga, O. (2009) Determination of the Concentration of Zinc and Vitamin C in Oysters and Some Medicinal Plants Used to Correct Male Factor Infertility. Journal of Natural Products, 2, 89-97.

[7]   Klotoe, J.R., Dougnon, T.V., Koudouvo, K., Ategbo, J.M., Koko, F., Akoegninou, A., Aklikokou, K., Dramane, K. and Gbeassor, M. (2013) Ethnopharmacological Survey on Antihemorrhagic Medicinal Plants in South of Benin. European Journal of Medicinal Plants, 3, 40-51.

[8]   Akinmoladun, A.C., Obutor, E.M. and Farombi, E.O. (2010) Evaluation of Antioxidant and Free Radical Scavenging Capacities of Some Nigerian Indigenous Medicinal Plants. Journal of Medicinal Food, 13, 444-451.

[9]   Eyong, K.O., Folefoc, G.N., Kuete, V., Beng, V.P., Krohn, K., Hussain, H., Nkengfack, A.E., Saeftel, M., Sarite, S.R. and Hoerauf, A. (2006) Newbouldiaquinone A: A Naphtoquinone-Anthraquinone Ether Coupled Pigment as a Potential Antimicrobial and Antimalarial Agent from Newbouldia laevis. Phytochemistry, 67, 605-609.

[10]   Amos, S., Binda, L., Vongtau, H., Chindo, B. and Abbah, J. (2002) Sedative Effects of the Methanolic Leaf Extract of Newbouldia laevis in Mice and Rats. Bollettino Chimico Farmaceutico, 141, 471-475.

[11]   Ainooson, G.K., Woode, E., Obiri, D.D. and Koffour, G.A. (2009) Antinociceptive Effects of Newbouldia laevis (P. Beauv) Stem Bark Extract in a Rat Model. Pharmacognosy Magazine, 5, 49-54.

[12]   Hassan, S.W., Salawu, K., Ladan, M.J., Hassan, L.G., Umar, R.A. and Fatihu, M.Y. (2010) Hepato-Protective, Antioxidant and Phytochemical Properties of Leaf Extracts of Newbouldia laevis. International Journal of PharmTech Research, 2, 573-584.

[13]   Kuete, V., Wabo, H.K., Eyon, K.O., Feussi, M.T., Wiench, B., Krusche, B. and Effert, T. (2011) Anticancer Activities of Six Selected Natural Compounds of Some Cameroonian Medicinal Plants. PLoS ONE, 6, e21762.

[14]   Bafor, E., Sanni, U. and Nworgu, Z.A. (2010) In Vitro Determination of the Mechanism of the Uterine Stimulatory Effect of Newbouldia laevis. Pharmaceutical Biology, 48, 808-815.

[15]   Awemu, G.A., Okunrobo, L.O. and Awah, F.M. (2012) Wound Healing and Antiulcer Activities of the Ethanol Extract of Newbouldia laevis Root Bark. Journal of Pharmacy & Bioresources, 9, 29-33.

[16]   Joppa, K.M., Vovor, A., Eklu-Gadegbeku, K., Agbonon, A., Aklikokou, K. and Gbeassor, M. (2008) Effect of Morinda lucida Benth. (Rubiaceae) and Newbouldia leavis P. Beauv. (Bignoniaceae) on Sickling of Red Blood Cells. Medecine Tropicale: Revue du Corps de Sante Colonial, 68, 251-256.

[17]   Owolabi, O.J., Amaechina, F.C. and Okoro, M. (2011) Effect of Ethanol Leaf Extract of Newbouldia laevis on Blood Glucose Levels of Diabetic Rats. Tropical Journal of Pharmaceutical Research, 10, 249-254.

[18]   Osigwe, C.C., Akah, P.A., Nworu, C.S., Okoye, T.C. and Tchimene, M.K. (2015) Antihyperglycemic Studies on the Leaf Extract and Active Fractions of Newbouldia laevis (Bignoniaceae). Pharmacology & Pharmacy, 6, 518-532.

[19]   Osigwe, C.C., Akah, P.A., Nworu, C.S. and Okoye, F.B.C. (2017) Apigenin: A Methanol Fraction Component of Newbouldia laevis Leaf, as a Potential Antidiabetic Agent. The Journal of Phytopharmacology, 6, 38-44.

[20]   Adesokan, A.A., Oyewole, O.I. and Turray Babara, M.S. (2009) Kidney and Liver Function Parameters in Alloxan Induced Diabetic Rats Treated with Aloe barbadensis Juice Extract. Sierra Leone Journal of Biomedical Research, 1, 33-37.

[21]   Onyije, F.M., Avwioro, O.G. and Waritimi, E.G. (2012) Evaluation of Nephrotoxic Effect of Bauhinia monandra on the Kidney of Alloxan-Induced Diabetic Rats. Journal of Pharmacy and Clinical Sciences, 4, 7-9.

[22]   Mansi, K. and Lehham, J. (2008) Effects of Artemisia sieberi Bessa a Herba-Alba on Heart Rate and Some Hematological Values in Normal and Alloxan-Induced Diabetic Rats. Journal of Basic and Applied Sciences, 4, 57-72.

[23]   Ajagbonna, O.P., Onifade, K.I. and Suleiman, U. (1999) Hematological and Biochemical Changes in Rats Given Extract of Calotropis procera. Sokoto Journal of Veterinary Sciences, 1, 36-42.

[24]   Yakubu, M.T., Akanji, M.A. and Oladiyi, A.T. (2007) Hematological Evaluation in Male Albino Rats Following Chronic Administration of Aqueous Extract of Fadogia agrestis Stem. Pharmacognosy Magazine, 3, 34-38.

[25]   National Institute of Health (NIH) (1985) Guide for the Use of Laboratory Animals DHHS, PHS NIH. Pub. No. 85-23.

[26]   Lorke, D. (1983) A New Approach to Acute Toxicity Testing. Archives of Toxicology, 54, 275-287.

[27]   Harbone, J.B. (1989) Phytochemical Methods. A Guide to Modern Technique of Plant Analysis. 3rd Edition, Chapman and Hill, London, 285.

[28]   Akah, P.A., Alemji, J.A., Salawu, O.A., Okoye, T.C. and Offiah, N.V. (2009) Effects of Vernonia amygdalina on Biochemical and Haematological Parameters in Diabetic Rats. Asian Journal of Medical Sciences, 1, 108-113.

[29]   Frode, T.S. and Medeiros, Y.S. (2008) Animal Models to Test Drugs with Potential Antidiabetic Activity. Journal of Ethnopharmacology, 115, 173-183.

[30]   Ezekwesili, C.N., Ogbunugafor, H.A. and Ezekwesili-Ofili, J.O. (2012) Antidiabetic Activity of Aqueous Extracts of Vitex doniana Leaves and Cinchona calisaya Bark in Alloxan-Induced Diabetic Rats. International Journal of Tropical Disease & Health, 2, 290-300.

[31]   Weatherburn, M.W. (1967) Urease-Berthelot Colorimetric Method for in Vitro Determination of Urea. Analytical Chemistry, 39, 971-974.

[32]   Bartels, H. and Bohmer, M. (1972) Quantitative Determination of Creatinine. Clinica Chimica Acta, 37, 193.

[33]   Jendrassik, L. and Grof, P. (1938) Simplified Photometric Methods for the Determination of Bilirubin. Biochemical Journal, 297, 81-89.

[34]   Rec, G.S.C. (1972) Colorimetric Method for Serum Alkaline Phosphatase Determination. Journal of Clinical Chemistry and Clinical Biochemistry, 10, 182-184.

[35]   Reitman, S. and Frankel, S. (1957) A Colorimetric Method for the Determination of Serum Glutamic Oxaloacetic and Glutamic Pyruvic Transaminases. American Journal of Clinical Pathology, 28, 56-63.

[36]   Gomall, A.G., Bardawill, C.S. and David, M.M. (1949) Determination of Serum Proteins by Means of the Biuret Reaction. The Journal of Biological Chemistry, 177, 751-766.

[37]   Miller, N.J., Rice-Evans, C., Davies, M.J., Gopinathan, V. and Milner, A. (1993) A Novel Method for Measuring Antioxidant Capacity and Its Application to Monitoring the Antioxidant Status in Premature Neonates. Clinical Science, 84, 407-412.

[38]   Kotharia, R. and Bokariya, P. (2012) A Comparative Study of Haematological Parameters in Type 1 Diabetes Mellitus Patients and Healthy Young Adolescents. International Journal of Biological and Medical Research, 3, 2429-2432.

[39]   Oyedemi, S.O., Adewusi, E.A., Aiyegoro, O.A. and Akinpeanolu, D.A. (2011) Antidiabetic and Hematological Effect of Aqueous Extract of Stem Bark of Afzelia africana (Smith) on Streptozocin-Induced Diabetic Wister Rats. Asian Pacific Journal of Tropical Biomedicine, 1, 353-358.

[40]   Arun, G.S. and Ramesh, K.G. (2002) Improvement of Insulin Sensitivity by Perindopril in Spontaneously Hypertensive and Streptozotocin Diabetic Rats. Indian Journal of Pharmacology, 34, 156-164.

[41]   Kolanjiappan, K., Manoharan, S. and Kayalvizhi, M. (2002) Measurement of Erythrocyte Lipids, Lipid Peroxidation, Antioxidants and Osmotic Fragility in Cervical Cancer Patients. Clinica Chimica Acta, 326, 143-149.

[42]   Mahmoud, A.M. (2013) Hematological Alterations in Diabetic Rats—Role of Adipocytokines and Effect of Citrus Flavonoids. EXCLI Journal, 12, 647-657.

[43]   Akomas, S.C., Okafor, A.I. and Ijioma, S.N. (2014) Glucose Level, Haematological Parameters and Lipid Profile in Ficus sur Treated Diabetic Rats. Comprehensive Journal of Agricultural and Biological Science, 2, 5-11.

[44]   Mohammed, A., Adelaiye, A.B., Bakari, A.G. and Mabrouk, M.A. (2009) Antidiabetic and Some Hematological Effects of Ethyl Acetate and n-Butanol Fractions of Ganoderma lucidium Aqueous Extract in Alloxan-Induced Diabetic Wistar Rats. International Journal of Medical Sciences, 1, 530-535.

[45]   Verma, N., Amresh, G., Sahu, P.K., Mishra, N., Singh, A.P. and Rao, C.V. (2012) Antihyperglycemic Activity, Antihyperlipidemic Activityy, Hematological Effects and Histopathological Analysis of Sapinus mukorossi Ganertan Fruits in Streptozocin Induced Diabetic Rats. Asian Pacific Journal of Tropical Medicine, 5, 518-522.

[46]   Ohlsson, A. and Aher, S.M. (2012) Early Erythropoietin for Preventing Red Blood Cell Transfusion in Preterm and/or Low Birth Weight Infants. The Cochrane Database of Systematic Reviews, No. 9, CD004863.

[47]   Abu-Zaiton, A.S. (2010) Antidiabetic Activity of Ferula asafoetida Extract in Normal and Alloxan Induced Diabetic Rats. Pakistan Journal of Biological Sciences, 13, 97-100.

[48]   Ogbonnia, S.O., Mbaka, G.O., Nkemehule, F.E., Emordi, J.E., Okpagu, N.C. and Ota, D.A. (2014) Acute and Subchronic Evaluation of Aqueous Extracts of Newbouldia laevis (Bignoniaceae) and Nauclea latifolia (Rubiaceae) Roots Used Singly or in Combination in Nigeria Traditional Medicine. British Journal of Pharmacology and Toxicology, 5, 55-62.

[49]   Kolawole, O.T., Akanji, M.A. and Akiibinu, M.O. (2013) Toxicological Assessment of Ethanolic Extract of the Leaves of Newbouldia laevis (P. Beauv). American Journal of Medicine and Medical Sciences, 3, 74-80.

[50]   Usman, H. and Osuji, J.C. (2007) Phytochemical and In-Vitro Antimicrobial Assay of the Leaf Extract of Newbouldia laevis. African Journal of Traditional, Complementary and Alternative Medicines, 4, 476-480.

[51]   Mahmoud, A.M., Ahmed, O.M., Ashour, M.B. and Abdel-Moneim, A. (2012) Hesperidin and Naringin Attenuate Hyperglycemia-Mediated Oxidative Stress and Proinflammatory Cytokine Production in High Fat Feed/Streptozocin Induced Type 2 Diabetic Rats. Journal of Diabetes and Its Complications, 26, 483-490.

[52]   Mahmoud, A.M., Ahmed, O.M., Ashour, M.B. and Abdel-Moneim, A. (2013) Upregulation of PPARγ Mediates the Antidiabetic Effects of Flavonoids in High Fat Diet Fed-Streptozocin Induced Type 2 Diabetic Rats. International Journal of Bioassays, 2, 756-761.

[53]   Ngogang, J.Y. (2005) Haematinic Activity of Hibiscus cannabinus. African Journal of Biotechnology, 4, 833-837.

[54]   Mohammed, R.K., Ibrahim, S., Atawodi, S.E., Eze, E.D., Suleiman, J.B. and Malgwi, I.S. (2013) Anti-Diabetic and Haematological Effects of n-Butanol Fraction of Alchornea Cordifolia Leaf Extract in Streptozotocin-Induced Diabetic Wistar Rats. Scientific Journal of Biological Sciences, 2, 45-53.

[55]   Uko, E.K., Erhabor, O., Isaac, I.Z., Abdulrahaman, Y., Adias, T.C., Sani, Y., Shehu, R.S., Liman, H.M., Dalltu, M.R. and Mainastra, A.S. (2013) Some Haematological Parameters in patiEnts with Type 1 Diabetes in Sokoto, North Western Nigeria. Journal of Blood & Lymph, 3, 2165-7831.

[56]   Tanigushi, A., Fukushima, M., Seino, Y., Sakai, M. and Yoshi, S. (2003) Platelet Count Is Independently Associated in Insulin Resistance in Non-Obese Japanese Type 2 Diabetic Patients. Metabolism, 52, 1246-1249.

[57]   Jesri, A., Okonufa, E.C. and Egan, B.M. (2005) Platelet and White Blood Cell Counts Are Elevated in Patients with the Metabolic Syndrome. The Journal of Clinical Hypertension (Greenwich), 7, 705-711.

[58]   Willams, W.J., Bentler, E., Erslev, A.J. and Lictman, M.A. (1983) Thrombocytosis. In: Heamatology, 3rd Edition, McGraw-Hill, New York, 1342-1344.

[59]   Edet, A.E., Patrick, E.B. and Olarufemi, E.A. (2013) Haematological Parameters of Alloxan-Induced Diabetic Rats Treated with Ethanol Extracts and Fractions of Nauclea latifolia Leaf. European Scientific Journal, 9, 203-210.

[60]   Rawi, S.M., Mourad, I.M. and Sayed, D.A. (2011) Biochemical Changes in Experimental Diabetes before and after Treatment with Mangifera indica and Psidium guava Extracts. International Journal of Pharmacy and Biomedical Sciences, 2, 29-41.

[61]   Gometi, S.A., Ogugua, V.N., Odo, C.E. and Joshua, P.E. (2014) Effects of Some Antidiabetic Plants on the Hepatic Marker Enzymes of Diabetic Rats. African Journal of Biotechnology, 13, 905-909.

[62]   Miikue-Yobe, T.F.B., Uwakwe, A.A. and Akaninwuor, J.O. (2015) Effect of Aqueous Leaf Extract of Heinisia crinata on Hematological and Some Biochemical Indices of Toxicity in Streptozocin Induced Diabetic Rats. International Journal of Innovation Science and Research, 2, 116-126.

[63]   Patrick, E.E., Item, J.A., Eyong, U.E. and Godwin, E.E. (2008) The Antidiabetic Efficacy of Combined Extracts from Two Continental Plants: Azadiracta indica and Vernonia amygdalina (African Bitter Leaf). American Journal of Biochemistry and Biotechnology, 4, 239-244.

[64]   Hasssan, S.W., Ayeni, G., Wasagu, R.S.U., Umar, R.A., Agaie, B.M. and Farouk, U.Z. (2015) Hepatoprotective Effect of Leaf Extracts of Newbouldia laevis on Carbon Tetrachloride-Intoxicated Rats. Journal of Molecular Global Biosciences, 5, 2258-2281.

[65]   Chen, H.-Y. and Yen, G.-C. (2007) Antioxidant Activity and Free Radical Scavenging Capacity of Extracts from Guava (P. guajava L.) Leaves. Food Chemistry, 101, 686-694.

[66]   Anaduaka, E.G., Ogugua, V.N., Chidozie, V. and Okonkwo, C.C. (2014) Ethanol Extracts of Newbouldia laevis Stem and Leaves Modulate Serum Marker Enzymes and Antioxidant Enzymes Activities in Diabetic Rats. African Journal of Biotechnology, 13, 2265-2272.

[67]   Imo, C., Uhegbu, F.O., Ifeanacho, N.G., Egbeigwe, O. and Ezekwe, A.S. (2014) Biochemical and Histopathological Changes Associated with Methanolic Leaf Extract of Gongronema latifolia in Acetaminophen-Induced Hepatic Toxicity in Wistar Albino Rats. International Journal of Biomolecules and Biomedicine, 4, 1-7.

[68]   Narasmihanaidu, K. and Ponnaian, S.M.P. (2006) Antihyperglycaemic and Antioxidant Effect of Rutin, a Polyphenolic Flavonoid in Streptozocin-Induced Diabetic Wistrar Rats. Basic & Clinical Pharmacology & Toxicology, 98, 97-103.

[69]   Yumnam, A.D., Vrushabendra, S.B.M., Vishwanath, S.K.M. and Ranu, R.R. (2012) Antidiabetic Activity of Echinochloa crusgalli (L.) P. Beauv Grains Extract in Alloxan Induced Diabetic Rats. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 3, 1257-1275.

[70]   Oyagbemi, A.A., Saba, A.B. and Arowolo, R.O.A. (2008) Safety Evaluation of Prolonged Administration of Stresroak in Grower Cockerels. International Journal of Poultry Science, 7, 574-578.

[71]   Shah, N.A. and Khan, M.R. (2014) Antidiabetic Effect of Sida cordata in Alloxan-Induced Diabetic Rats. BioMed Research International, 2014, Article ID: 671294.

[72]   Rana, S., Singh, R. and Verma, S. (1996) Protective Effects of Few Antioxidants on Liver Function in Rats Treated with Cadmium and Mercury. Indian Journal of Experimental Biology, 34, 177-179.

[73]   Sunmonu, T.O. and Afolayan, A.J. (2013) Evaluation of Antidiabetic Activity and Associated Toxicity of Artemisia afra Aqueous Extract in Wistar Rats. Evidence-Based Complementary and Alternative Medicine, 2013, Article ID: 929074.

[74]   Alarcon, C., Wicksteed, B., Prentki, M., Corkey, B.E. and Rhodes, C.J. (2002) Succinate Is a Preferential Metabolic Stimulus-Coupling Signal for Glucose-Induced Proinsulin Biosynthesis Translation. Diabetes, 51, 2496-2504.

[75]   Abdulazeez, M.A., Ibrahim, K., Bulus, K., Babvoshia, H.B. and Abdullahi, Y. (2013) Effect of Combined Use of Ocimum gratissimum and Vernonia amygdalina Extract on the Activity of Angiotensin Converting Enzyme, Hypolipidemic and Antioxidant Parameters in Streptozocin-Induced Diabetic Rats. African Journal of Biochemistry Research, 7, 165-173.

[76]   Antu, K.A., Riya, M.P., Mishra, A., Anilkumar, K.S., Chandrakanth, C.K., Tamrakar, A.K., Srivastava, A.K. and Raghu, K.G. (2014) Antidiabetic Property of Symplocos cochinchinensis Is Mediated by Inhibition of Alpha Glucosidase and Enhanced Insulin Sensitivity. PLoS ONE, 9, e105829.

[77]   Cao, G., Booth, S.L., Sadowski, J. A. and Prior, R.L. (1998) Increase in Human Plasma Antioxidant Capacity Following Consumption of Controlled Diets High in Fruits and Vegetables. The American Journal of Clinical Nutrition, 68, 1081-1087.

[78]   Akinosun, O.M. and Bolajoko, E.B. (2007) Total Antioxidant Status in Type 2 Diabetic Patients: Experience at University College Hospital (UCH) Ibadan Nigeria. Nigerian Journal of Clinical Practice, 10, 126-129.

[79]   Saeed, M.K., Dend, Y. and Dai, R. (2008) Attenuation of Biochemical Parameters in Streptozocin-Induced Diabetic Rats by Oral Administration of Extracts and Fractions of Cephalotaxus sinensis. Journal of Clinical Biochemistry and Nutrition, 42, 21-28.