|Year : 2013 | Volume
| Issue : 1 | Page : 43-47
Subacute effect of Vernonia amygdalina methanolic crude leaf extract on the hematological profile in albino Wistar rats
Thomas Nubila1, Nkoyo Imelda Nubila2, Ayodele Ukaejiofo3, Elvis Neba Shu2, Chukwubuzor U Okwuosa4, Chioli Pascal Chijioke2, Princess Orji1
1 Department of Medical Laboratory Sciences, Faculty of Medical Sciences, College of Medicine, University of Nigeria, Enugu Campus, Enugu, Enugu State, Nigeria
2 Department of Pharmacology, and Therapeutics, Faculty of Medical Sciences, College of Medicine, University of Nigeria, Enugu Campus, Enugu, Enugu State, Nigeria
3 Department of Medical Rehabilitation, Faculty of Health Sciences and Technology, University of Nigeria Teaching Hospital (UNTH), Ituku Ozalla, Enugu, Enugu State, Nigeria
4 Department of Histopathology, University of Nigeria Teaching Hospital (UNTH), Ituku Ozalla, Enugu, Enugu State, Nigeria
|Date of Web Publication||30-Dec-2013|
Department of Medical Laboratory Sciences, University of Nigeria, Enugu Campus, Enugu State
Source of Support: None, Conflict of Interest: None
Background: Vernonia amygdalina is among one of the most used medicinal plants with a higher safety and efficacy in managing a wide array of medical ailments. The objective of this study is to evaluate the subacute effects of the methanolic crude leaf extract of Vernonia amygdalina on the hematological profile in Albino Wistar rats. Materials and Methods: Thirty Albino rats were used for the study. They were equally grouped into five groups: A (control), B, C, D, and E groups. The test (B, C, D, and E) groups were gavaged with doses (10, 15, 25, and 40 mg/kg body weight, respectively) of Vernonia amygdalina leaf methanolic extract once daily, for nine days. Two milliliters of blood was collected from two animals each from each group on days three, six, and nine, for the determination of the hematological profile using a hematology analyzer. Results: Mean cell hemoglobin mean value showed a statistically significant increase (P < 0.05) in group C (15 mg/kg body weight). Similarly, a statistically significant increase (P < 0.01) was recorded in group E (40 mg/kg body weight) on day three, when compared with the control group A (untreated). However, only the mean value of the platelet count revealed a statistically significant decrease (P < 0.05) in group D (25 mg/kg body weight) on day six, when compared with the control. Conclusion: The subacute administration of Vernonia amygdalina methanolic crude leaf extract does not possess hematopoietic activities.
Keywords: Hemopoiesis, leaf extract, subacute, Vernonia amygdalina
|How to cite this article:|
Nubila T, Nubila NI, Ukaejiofo A, Shu EN, Okwuosa CU, Chijioke CP, Orji P. Subacute effect of Vernonia amygdalina methanolic crude leaf extract on the hematological profile in albino Wistar rats. Niger J Exp Clin Biosci 2013;1:43-7
|How to cite this URL:|
Nubila T, Nubila NI, Ukaejiofo A, Shu EN, Okwuosa CU, Chijioke CP, Orji P. Subacute effect of Vernonia amygdalina methanolic crude leaf extract on the hematological profile in albino Wistar rats. Niger J Exp Clin Biosci [serial online] 2013 [cited 2023 Mar 30];1:43-7. Available from: https://www.njecbonline.org/text.asp?2013/1/1/43/123963
| Introduction|| |
It is estimated that more than 40% of the prescribed medicines in the western world, even today, come directly or indirectly from plants.  In the developing countries of Africa, such as Nigeria, Cameroon, Kenya, Zimbabwe, and Malawi, the application of medicinal plants, especially in traditional medicines, is currently a well-acknowledged and established or viable profession. This is practically so in rural communities, where the services of modern healthcare may be limited or unaffordable. Medicinal plants are, therefore, an important source of self-reliance in tropical countries. 
Of late, there has been a great deal of interest worldwide in the role of complementary and alternative medicines for the treatment of various acute and chronic diseases,  especially in the developing countries. Of the various classes of phytochemicals, interest has been focused on the anti-inflammatory and antioxidant properties of polyphenols found in different botanical substances. Plants, vegetables, and spices used in folk and traditional medicine for nutritional and medicinal purposes have gained wide acceptance as one of the main sources in prophylactic and chemopreventive drug discoveries and development.  Indeed, most parts of plants are rich sources of phenolic and other related compounds and have been recognized to possess a wide range of potentials, including, antioxidant, antibacterial, anti-inflammatory, hepatoprotective, hematinics, and anti-carcinogenic actions. Many of the biological functions of flavonoids, phenolic compounds, and other complex substances have been attributed to their free radical scavenging, metal ion chelating, and antioxidant activities. ,
Vernonia amygdalina, commonly known as bitter leaf, is a shrub that grows up to three meters high in the African tropics and other parts of Africa, particularly, Nigeria, Cameroon, and Zimbabwe.  It is reputed to have several health and nutritional benefits. The organic fraction extracts of the plant are seen to possess cytotoxic effects toward human carcinoma cells of the nasopharynx.  They are effective against amoebic dysentery,  gastrointestinal disorders,  and have antimicrobial and antiparasitic activities. , The biologically-active compounds of Vernonia amygdalina are saponins and alkaloids,  terpenes, steroids, coumarins, flavonoids, phenolic acids, lignans, xanthones and anthraquinones,  edotides,  and sesquiterpenes.  Vernonia amygdalina has been shown to contain appreciable quantities of ascorbic acid and carotenoids. , Calcium, iron, potassium, phosphorus, manganese, copper, and cobalt have also been found in significant quantities in Vernonia amygdalina. ,,
Hemopoiesis refers to the formation of blood, particularly the mechanism responsible for the formation and maturation of red cells (erythrocytes), white cells (leucocytes), and platelets (thrombocytes). , In healthy individuals there is a constant balance between the rate of formation and destruction of these cells. Disruption of this critical balance, either in terms of changes in the rate of formation, disordered formation or increased utilization and destruction is essentially responsible for the final pathological state.  Red cell disorders are usually reflected in anemia. White cell disorders, in contrast, most often involve overgrowth, usually malignant. Hemorrhagic diathesis is the result of hemostatic derangement. 
Despite the wide and large consumption of Vernonia amygdalina leaves for either nutritional or medicinal purposes in the sub-Sahara countries, its influence on the hematological parameters has not been evaluated. The aim of this study, therefore, is to determine the subacute effects of the crude methanolic seed extract of Vernonia amygdalina on the hematological profile in male Albino Wistar rats.
| Materials and Methods|| |
Thirty (30) male, white Albino Wistar rats, three to four months old, and weighing 120-160 g were purchased from an Animal House in the University of Nigeria, Enugu Campus, in Enugu State, Nigeria. They were grouped into five groups, A, B, C, D, and E, according to similar body weight. Group A served as the control and received no treatment. The animals were fed and acclimatized for one week. They were handled according to the University of Nigeria Nsukka, Enugu State, Nigeria, Animal Study Review Board approved guidelines for the 'care and use of laboratory animals'.
Plant Identification, Extraction, and Administration
A sample of the leaf of V. amygdalina was authenticated at the Herbarium Section of the Department of Botany, University of Nigeria, Nsukka, Enugu State, Nigeria, by a taxonomist. A voucher specimen (UNH N N° 308 b ) was deposited at the herbarium for future reference. The leaf was bought from the Ogbete market in Enugu, Enugu State, Nigeria. After collection and washing with clean tap water, it was air-dried under shade for six days. This was ground into fine powder. Four hundred grams of the fine powder was dissolved in 2000 ml of 98% methanol as a solvent. This was shaken vigorously twice daily, for two days. The extract was strained using fine cheese cloth. The filtrate was then filtered using Whatman filter paper No 1. This was air-dried by evaporation in an open stainless metallic basin at room temperature. The semi-solid green residue was reconstituted using distilled water in graded doses (10, 15, 25 and 40 mg/kg body weight). This was orally administered to the test groups (B, C, D, and E), respectively, once daily for nine days.
Sample Collection and Analysis
Two milliliters of venous blood was painlessly collected, aseptically, from the retrobulbar plexus of the median canthus of the eye on days three, six, and nine after oral methanolic crude leaf administration, under thiopentone sodium anesthesia.  This was transferred into a tripotassium Ethylenediaminetetraacetic acid (K 3 EDTA) anticoagulant bottle. This was mixed well immediately by gentle inversion and used for the determination of the hematological parameters, using an automated hematology autoanalyzer (Sysmex KX-21N), following the manufacturers' guidelines. All the samples were analyzed within two hours of collection.
Data analysis was done using the GraphPad prism computer software. Students 't'-test and one-way analysis of variance (ANOVA) were used for comparison. A P-value < 0.05 was considered significant.
| Results|| |
There was no statistically significant difference (P > 0.05) in the three major blood cells (erythrocytes, leucocytes, and thrombocytes) in all the test groups, when compared with the control group [Table 1] and [Table 3]. The mean value of the mean cell hemoglobin showed a statistically significant increase (P < 0.05) in group C (15 mg/kg body weight). Similarly, a statistically significant increase (P < 0.01) was recorded in group E (40 mg/kg body weight) on day three, when compared with the control group A (untreated) [Table 1]. However, only the platelet count mean value revealed a statistically significant decrease (P < 0.05) in group D (25 mg/kg body weight) on day six, compared to the control [Table 2]. In addition, none of the parameters evaluated showed a statistically significant difference (P > 0.05) in the entire test groups when compared with the control group on day nine.
|Table 1: Comparison of the mean ± SE of the hematological parameters with the control group after three days of oral crude extract administration|
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|Table 2: Comparison of the mean ± SE of the hematological parameters with the control group after six days of oral crude extract administration|
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|Table 3: Comparison of the mean ± SE hematological parameters with the control group after nine days of oral crude extract administration|
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| Discussion|| |
Medicinal plants have played a key role in the world healthcare with about 80% of the Africans depending on phytomedicine, which has shown a wide range of uses in the treatment of diseases especially priority diseases of Africa such as human immunodeficiency virus infection/acquired immunodeficiency syndrome (HIV/AIDS), malaria, sickle-cell anemia, diabetes, and hypertension. These medicinal plants have a more beneficial effect than their synthetic counterparts by being safer, acceptable, affordable, culturally compatible, and suitable for chronic treatment. 
From the results of the present study, there was a marginal general increase, although not statistically significant (P > 0.05), in the major blood cells (red blood cells, white blood cells, and the platelets) in all the test groups, when compared with the control group, throughout the study period. This indicates that the crude leaf methanolic extract of Vernonia amygdalina is most likely to stimulate hemopoeisis, maybe in a chronic study. Also these hemopoietic activities can be attributed to the rich phytochemical compositions that include proteins, ,,, carbohydrates, ,, iron, , and most probably the B Vitamins, which are all required for the regulation and synthesis of blood cells (hemopoeisis). Even though the mechanism is unknown, this process can be achieved through the stimulation of cytokines such as interleukin 6 (IL-6) or growth factors such as erythropoietin (EPO) on the receptors on the pluripotent progenitor stem cells in the bone marrow microenvironment. However, there is no previous study or literature for comparison, to support these findings. This result agrees with that of Momoh et al., who has found no significant changes in the hematological parameters between the treated and control groups indicating that V. amygdalina is not hematotoxic at lower doses.
In conclusion, the Vernonia amygdalina methanolic crude leaf extract does not possess subacute hemopoietic activity and is not hematotoxic.
| References|| |
|1.||Okafor JC, Amaechi NO. Importance of tropical plants. In: The Association for Scientific Identification, Conservation and Utilization of Medicinal Plants of Nigeria. 1 st ed. Checklist of medicinal plants of Nigeria and their uses. Nigeria: Jamoe Publishers; 2005. p. 1-5. |
|2.||Aruoma OI, Sun B, Fujii H, Neergheen VS, Bahorun T, Kang KS, et al. Low molecular proanthocyanidin dietary biofactor Oligonol: Its modulation of oxidative stress, bioefficacy, neuroprotection, food application and chemoprevention potentials. Biofactors 2006;27:245-65. |
|3.||Seeff LB, Lindsay KL, Bacon BR, Kresina TF, Hoofnagle JH. Complementary and alternative medicine in chronic liver disease. Hepatology 2001;34:595-603. |
|4.||Winslow LC, Kroll DJ. Herbs as medicines. Arch Intern Med 1998;158:2192-9. |
|5.||Ifeoma II, Chukwunonso EC. Current perspectives on the medicinal potentials of Vernonia amygdalina Del. J Med Plant Res 2011;5:1051-61. |
|6.||Kupchan SM, Hemnigway RJ, Karim A, Werner D. Tumor inhibitors. XLVII. Vernodalin and vernomygdin, two new cytotoxic sesquiterpene lactones from Vernonia amygdalina Del. J Org Chem 1969;34:3908-11. |
|7.||Moundipa FP, Flore MG, Bilong FC, Bruchhaus I. In vitro amoebicidal activity of some medicinal plants of the Bamun region (Cameroon). Afr J Tradit Compliment Altern Med 2005;2:113-121. |
|8.||Akah PA, Ekekwe RK. Ethnopharmacology of some asteraceae family used in Nigerian traditional medicine. Fitoterapia 1995;66:351-5. |
|9.||Akinpelu DA. Antimicrobial activity of Vernonia amygdalina leaves. Fitoterapia 1999;70:432-4. |
|10.||Hladik C, Krief S, Haxaire C. Ethnomedicinal and bioactive properties of plants ingested by wild chimpanzees in Uganda. J Ethnopharmacol 2005;101:1-15. |
|11.||Muraina IA, Adaudi AO, Mamman M, Kazeem HM, Picard J, McGaw LJ, et al. Antimycoplasmal activity of some plant species from northern Nigeria compared to the currently used therapeutic agent. Pharm Biol 2010;48:1103-7. |
|12.||Cimanga RK, Tona L, Mesia K, Musuamba CT, De Bruyne T, Apers S, et al. In vitro anti- plasmodial activity of extracts and fractions from seven medicinal plants used in the Democratic Republic of Congo. J Ethnopharmacol 2004;93:27-32. |
|13.||Izevbigie EB. Discovery of water-soluble anticancer agents (edotides) from a vegetable found in Benin City, Nigeria. Exp Biol Med 2003;228:293-8. |
|14.||Udensi EA, Ijeh II, Ogbonna U. Effect of traditional processing on the phytochemical and nutrient composition of some local Nigerian leafy vegetables. J Sci Tech 2002;8:37-40. |
|15.||Ejoh RA, Nkonga DV, Innocent G, Moses MC. Nutritional components of some non-conventional leafy vegetables consumed in Cameroon. Pak J Nutr 2007;6:712-7. |
|16.||Bonsi ML, Osuji PO, Tuah AK, Umunna MN. Vernonia amygdalina as supplement of teff straw (Eragrostis tef) fed to Ethiopian Menz sheep. Agroforestry Syst 1995;31:229-41. |
|17.||Eleyinmi AF, Sporns P, Bressler DC. Nutritional composition of Gongronema latifolium and Vernonia amygdalina. Nutr Food Sci 2008;38:99-109. |
|18.||Orkin SH. Hematopoiesis: How does it happen? Curr Opin Cell Biol 1995;7:870-7. |
|19.||Orkin SH. Development of the hematopoietic system. Curr Opin Genet Dev 1996;6:597-602. |
|20.||Raphel SS, Culling CF, Hyde TA. In: Wood MJ, Mellor LD, Sergovich F, Thompson S, editors. Lynch's Medical Laboratory Technology. 3 rd ed. Philadelphia: WB Saunders Co.; 1976. p. 782-3. |
|21.||Kumar V, Cotran RS, Robbins SL. Basic Pathology. 6 th ed. Philadelphia: WB. Saunders; 1985. p. 1. |
|22.||Parasuraman S, Raveendran R, Kesavan R. Blood sample collection in small laboratory animals. J Pharmacol Pharmacother 2010;2:87-93. |
|23.||Okigbo RN and Mmeka EC. An appraisal of phytomedicine in Africa. KMITL Sci Tech J 2006;6:83-94. |
|24.||Igile GO, Oleszek W, Jurzysta M, Burda S, Fafunso M, Fasanmade AA. Flavonoids from Vernonia amygdalina and their antioxidant activities. J Agric Food Chem 1994;42:2445-8. |
|25.||Momoh MA, Adikwu MU, Ibezim EC and Attama AA. Effect of metaformin and Vernonia amygdalina leaf extract loaded PEGylated - mucin formulation an hematological, kidney and liver indices of healthy and Diabetes rats. J Pharm Res 2011;4:3455-9. |
[Table 1], [Table 2], [Table 3]