|Year : 2014 | Volume
| Issue : 1 | Page : 54-58
Effects of aqueous leaf extract of azadirachta indica on some haematological parameters and blood glucose level in female rats
Eghosa E Iyare, Nancy N Obaji
Department of Physiology, College of Medicine, University of Nigeria, Enugu Campus, Nigeria
|Date of Web Publication||1-Jul-2014|
Eghosa E Iyare
Department of Physiology, College of Medicine, University of Nigeria, Enugu Campus
Source of Support: None, Conflict of Interest: None
Background: Azadirachta indica A. juss (AI; Family: Meliaceae) is one of the most useful medicinal plants containing different alkaloids that includes nimbitin, azadirachtin and salanin whose individual effects contribute to the general medicinal properties of the plant. In Nigeria, pregnant and lactating women have been observed consuming aqueous extract of AI and anecdotal reports from them suggest that they consume the extract because of the folkloric belief that it is potentially harmless, it is hematopoietic and protects them from malaria. Aim: This study was designed to investigate the effects of consumption of AI during pregnancy and lactation on some hematological parameters and blood glucose level. Materials and Methods: Sixty female rats weighing 150-200 g were used for this study. The rats were divided into three groups (non pregnant, pregnant and lactating groups) of 20 rats each. Each group was subdivided into four subgroups based on the dose of AI administered. Subgroup I served as control and received distilled water throughout the experiment while subgroups II-IV received 200 mg/kg, 400 mg/kg and 600 mg/kg body weight of the extract respectively for 21 days. On day 20 of extract administration, blood samples were withdrawn from each rat in each group after an overnight fast for the estimation of some hematological parameters and blood glucose level. Results: There were significant increases (P < 0.05 for each) in the packed cell volume (PCV), hemoglobin concentration (Hb), red blood cell (RBC), white blood cell (WBC), and platelet (PLT) and a significant dose-dependent decrease (P < 0.05) in blood glucose level in all groups. Conclusion: It is concluded that the results of the present study seem to justify the folkloric use of AI as a hematopoietic agent with the potential of ameliorating the burden of anemia and hyperglycemia in women especially during pregnancy.
Keywords: Azadirachta indica, anemia, blood glucose, haematological parameters
|How to cite this article:|
Iyare EE, Obaji NN. Effects of aqueous leaf extract of azadirachta indica on some haematological parameters and blood glucose level in female rats. Niger J Exp Clin Biosci 2014;2:54-8
|How to cite this URL:|
Iyare EE, Obaji NN. Effects of aqueous leaf extract of azadirachta indica on some haematological parameters and blood glucose level in female rats. Niger J Exp Clin Biosci [serial online] 2014 [cited 2019 Aug 21];2:54-8. Available from: http://www.njecbonline.org/text.asp?2014/2/1/54/135731
| Introduction|| |
Since ancient times, plants have played key roles in traditional health care systems and also form the basis of a significant percentage of allopathic and modern drugs in industrialized nations of the world. , In Africa alone, up to 80% of the population uses traditional medicine for primary healthcare and in industrialized parts of the world its counterpart or adaptations also exist in various forms called Complementary and Alternative Medicine (CAM).  The medicinal values of these plants used in traditional medicine lie in some complex chemical substances that produce a definite physiological action on the human body. 
Azadirachta indica A. juss (AI; Family: Meliaceae) is a popular medicinal plant originally grown in India  but is now being cultivated in almost every part of the world including Nigeria , where it is popularly called "Dogonyaro". It is one of the most useful medicinal plants.  It is a large evergreen tree growing 10-11 m tall. The leaves are divided into numerous leaflets each resembling a full-grown leaf. , The plant contains different alkaloids which include; nimbitin, azadirachtin and salanin  , whose individual effects contribute to the general medicinal properties of the plant.
In traditional medicine a decoction made from the bark, leaf, root, fruits and flowers is used in the treatment of a variety of ailments including blood morbidity, biliary afflictions, and itching, skin as well as peptic ulcers. ,
Apart from their traditional uses, there are several reports on the biological activities and pharmacological actions of AI based on modern scientific investigations. ,,,,,, Blood glucose lowering activity of AI seed oil and leaf extracts have been reported in various models of diabetic animals. ,,, Ethanol extracts of AI leaves have been shown to demonstrate anti-lipid peroxidative, anti-hyperglycemic and anti-hypercholesterolemic activites as well as reduced serum triglyceride level in diabetic rat model.  Also, a significant decrease in some hematological parameters in chickens fed with AI leaves  and no significant difference in some hematological parameters following the extract use in diabetic rats  have been reported.
In Nigeria decoctions and aqueous extract of AI are commonly used in the treatment of malaria. Some women have been observed consuming aqueous extract of AI during pregnancy and lactation and anecdotal reports from them suggest that they consume the extract because of the belief that it is potentially harmless, it is hematopoietic and protects them from malaria. There is however, paucity of data on the effects of consumption of AI during pregnancy and lactation on some hematological parameters and blood glucose levels. The present study was designed to investigate whether or not the consumption of aqueous AI during pregnancy and lactation would affect hematological parameters and blood glucose level in rats.
| Materials and methods|| |
Plant Materials and Extract Preparation
Fresh matured leaves from the AI tree located at the University of Nigeria, Enugu campus were harvested and identified by a botanist, Prof. M. O. Nwosu, at the University of Nigeria where a voucher specimen (UNH No. 521 A ) was deposited for further reference. The leaves were then washed and air-dried. The dried leaves were homogenized using an electric blender. The powder was then exhaustively extracted at 60 o C for 8 hours, using soxhlet extractor according to the method of Biu et al.  The extract was then concentrated in a water bath (40 o C) yielding 340 g of brown substance which was then stored in a refrigerator (4 o C) until use. When needed, this was reconstituted in distilled water to give the required doses of 200, 400, and 600 mg/kg body weight used in the present study. Water was used in the extraction as against ethanol and acetone-water  , because we wanted to simulate as close as possible the mode of preparation locally.
Experimental Animals and Extract Administration
Sixty matured virgin female rats weighing between 150-200 g were used for this study. The rats were housed in well-ventilated cages and acclimatized for 3 weeks in the animal house of the Department of Physiology under controlled environmental conditions. The animals were provided standard rat pellet feed and tap water ad libitum. The rats were randomly divided into three broad groups (nonHpregnant, pregnant, and lactating groups) of 20 rats each. Each group was further subdivided into four subgroups depending on the dose of AI administered. In the non-pregnant group, the rats were subdivided into the four subgroups as stated above immediately after acclimatization and the extract administration commenced immediately. In the pregnant group, the estrus cycle was monitored for each rat by examining the daily vaginal smears under light microscopy. At proestrus, male rats of proven fertility were introduced into female cages to allow mating. Mating was proved successful when spermatozoa were observed in the vaginal smear of the female rats the following morning and this was regarded as day 1 of pregnancy.  On day 1 of pregnancy, the rats were then subdivided into four subgroups as stated above and the extract administration commenced immediately. For the lactation group, the rats were subdivided into four subgroups as stated above on the day of delivery and the extract administration also commenced.
Subgroup I rats served as the control group and received distilled water throughout the experiment while subgroups II-IV rats received 200 mg/kg, 400 mg/kg and 600 mg/kg body weight of the extract respectively by gavage using an oral cannula with blunt curved end for 21 days. On day 20 of extract administration, blood samples were withdrawn from the orbital sinus of each rat in each group after an overnight fast, according to the method of Raji et al., for estimation of blood glucose level and some hematological parameters.
Fasting blood glucose level was estimated by glucose oxidase-peroxidase reactive strips (Accu-check, Roche Diagnostic, USA) while the blood parameters; packed cell volume (PCV), Hemoglobin Concentration (Hb), white blood cell count (WBC), red blood cell count (RBC) and platelet count (PLT); were analyzed using an Automated Hematologic Analyzer (Sysmex, KX-21, Japan).
All procedures used in this study conformed to the guiding principles for research involving animals as recommended by the Declaration of Helsinki and the Guiding principles in the Care and Use of animals. 
Data were expressed as means ± standard error of mean (M ± SEM) and were subjected to one way analysis of variance (ANOVA) followed by a post hoc Students' Neuman-Keuls test. Statistical significance was considered at P < 0.05.
| Results|| |
AI significantly increased (P < 0.05 for each) the PCV, Hb, RBC, and WBC values compared with control values at all doses investigated [Table 1]. AI at 200 mg/Kg did not significantly increase (P > 0.05) the RBC value compared with control. The platelets were not significantly different (P > 0.05) among the various groups. The blood glucose level was significantly reduced (P < 0.05 for each) at all doses of AI investigated when compared with the control.
|Table 1: Effect of aqueous extract of AI on hematological parameters and blood glucose level in non-pregnant rats|
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AI at all doses investigated significantly increased (P < 0.05 for each) all blood parameters (PCV, Hb, RBC, WBC, and platelets) measured [Table 2]. AI at all doses investigated also significantly decreased (P < 0.05 for each) the blood glucose level.
|Table 2: Effect of aqueous extract of AI on hematological parameters and blood glucose level in pregnant rats|
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AI at 200 mg/kg did not significantly affect (P > 0.05) the blood parameters measured whereas the 400 mg/kg and 600 mg/kg significantly increased (P < 0.05 for each) the blood parameters measured with the effects on PCV and RBC being dose-dependent [Table 3]. AI at the doses investigated significantly decreased (P < 0.05 for each) the blood glucose level in a dose-dependent manner.
|Table 3: Effect of aqueous extract of AI on hematological parameters and blood glucose level in lactating rats|
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| Discussion|| |
The female rats treated with aqueous leaf extract of AI at different stages of reproductive life (non pregnant, pregnant and lactation) showed an increase in the PCV, Hb, RBC, and WBC. This observation is in agreement with the report of Parshad et al. Also, Mbah et al,. working with HIV/AIDS patients, also observed that an acetone/water leaf extract of AI (IRAB) showed a significant increase in some hematological parameters. Since pregnancy is a potentially anaemic state , and pregnancy is often complicated by malaria in Nigeria, this observed increase in hematological parameters in the present study suggests that the extract may have potentials at ameliorating the burden of anaemia in pregnancy. Indeed, this may justify the folkloric use of this plant during pregnancy. The significance of the observed increased hematological parameters in the present study cannot be over emphasised as anemia has been reported to affect over 500 million women  and in pregnancy it is associated with impaired maternal and infant outcome.
The increased blood parameters could be related to the reported constituents of the extract (flavonoids and quercetin) that have been shown to have hematopoietic properties.  Also, AI has been reported to boost the body's macrophage response, which stimulates the body's lymphatic system and also boosts the body's production of WBCs. ,
The effect of AI at decreasing the blood glucose level in normal and experimentally induced diabetic animals is well established. ,, The results of the present study also showed that the extract significantly lowered the blood glucose level in these rats. This suggests that AI at the doses tested, also possess glucose lowering effect in the normal non-pregnant, pregnant and lactating state. Several mechanisms through which AI decreases the blood glucose level have been suggested by several authors. , Jelodar et al., suggested that the glucose-lowering properties of the extract may be related to the ability of the extract to stimulate sufficient production of insulin by the pancreas, that aided in the peripheral utilization of glucose in the cells or the possible ability of the extract to regenerate the β-cells. Chattopadhyay et al., on the other hand suggested that AI's possible mechanism is by inhibiting the action of epinephrine on glucose metabolism resulting in increased utilization of peripheral glucose.
In conclusion, the results of the present study seem to justify the folkloric use of AI as a hematopoietic agent with the potential of ameliorating the burden of anemia and hyperglycemia in women especially during pregnancy.
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[Table 1], [Table 2], [Table 3]
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