|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 On day 20 of extract administration, blood samples were withdrawn from each rat in each group after an overnight fast for the estimation of some hematological parameters and blood glucose level.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 Mar 23];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.
| References|| |
|1.||Calson IJ. Ethnomedical field research, medicinal plants and tropical public health. Rainforest Med Bull 1998;5:8-15. |
|2.||Samy RP, Gopalakrishnakone P. Current status of herbals and their future perspective. Nature Proceeding: 2007. Available from: http://hdl.handle.net/10101/npre.2007.1176.1 [Last accessed on 2014 Mar 29]. |
|3.||WHO (World Health Organisation) (2003). Traditional Medicine. Available from: www.who.int/media centre/fact sheet/fs134/en/ [Last retrieved 2008 Jun 9]. |
|4.||Nubila T, Ukaejiofo EO, Nubila NI, Shu EN, Okwuosa CU, Ukaejiofo AC, et al. Effects of methanolic seed extract of Telfairia occidentalis on blood coagulation in Albino rats. Niger J Exp Clin Biosci 2013;1:10-3. |
|5.||Ofem OE, Ikpi DE, Essien NM. Increased bile flow rate and altered composition of bile induced by ethanolic leaf extract of Azadirachta indica (neem) in rats. Niger J Exp Clin Biosci 2013;1:18-22. |
|6.||Das BK, Mukherjee SC, Murjani O. Acute toxicity of neem (AI) in Indian carps. J Aquac Trop 2003;17:23-33. |
|7.||Sonibare MO, Isiaka AO, Taruka WM, Williams NS, Soladoye M, Emmanuel O. Constituents of neemleaves. Nat Prod Commu 2006:23-6. |
|8.||Kausik BI, Ranajet KB. Biological activities and medicinal properties of neem plant. Curr Sci 2002;82:1336-45. |
|9.||Koul O, Isman MB, Ketkar CM. Properties and uses of neem, Azadirachta indica. Can J Bot 1990;68:1-11. |
|10.||Chatterjee A, Pakrashi S, Eds. The treatise on Indian medicinal plants. New Delhi: Publications and Information Directorates; 1994. p. 76. |
|11.||Mitra CR, Patel MS. Neem. Indian Central oil seeds committee, Hyderabad; 1963; p. 69-94. |
|12.||Schumutterer H. The Neem Tree, source of unique Natural producs for intergrated pest management, medicine, industry and other purposes. 2 nd ed. Germany: VCH Weinhein; 2002. p. 68-76. |
|13.||Dixit VP, Sihn R, Tarok R. Effect of neem seed oil on the blood glucose concentrationof normal and alloxan diabetic rats. J Ethnophamacol 1986;17:95-8. |
|14.||Halim EM. Lowering of blood sugar by water extract of Azadirachta indica and Abroma augusta in diabetic rats. Indian J Exp Biol 2003;41:636-40. |
|15.||Gupta S, Kafaria M, Gupta PK, Murganandan S, Yashroy RC. Protective role of extracts of neem seeds in diabetes caused by streptozotocin in rats. J Ethnopharmacol 2004;90:185-9. |
|16.||Khosla P, Bhanwra S, Singh J, Seth S, Srivastava RK. A study of hypoglycaemic effects of Azadirachta indica (neem) in normal and alloxan diabetic rabbits. Indian J Physical Pharmacol 2008;44:69-74. |
|17.||Ekaidem IS, Akpan HD, Usoh IF, Etim OE, Ebong PE. Effects of ethanolic extract of Azadirachta indica leaves on lipid peroxidation and serum lipids of diabetic Wistar rats. Acta Biol Szegedensis 2007;51:17-20. |
|18.||Biu AA, Yusufu SD, Rabo JJ. Acute toxicity study on neem (azadirachta indica, Juss) leaf aqueous extract in chicken (gallus gallus domesticus). Afr Sci 2010;11. |
|19.||Itemobong SE, Atanghwo IJ, Akapan HD, Usoh IU, Etim OE, Ebong PE. Effects of ethanol extract of azadirachta indica on some immunological and haematological parameters of diabetic wistar rats. Afr J Pharm Pharmacol 2010;4:104-8. |
|20.||Mallie JP, Boudzoumou P. Functional renal maturation in rats neonates after prenatal exposure to furosemide. Pediatr Nephrol 1996;10:458-60. |
|21.||Raji Y, Morakinyo AO, Oloyo AK, Akinsomisoye OS, Kunle-Alabi OT, Esegbue PR, et al. Impact of chloroform extract of Carica papaya seeds on estrous circle and fertility in female albino rats. J Med Sci 2005;5:337-43. |
|22.||American Physiological Society. Guiding principles for research involving animals and human beings. Am J Physiol 2002;283:R281-3. |
|23.||Parshad O, Singh P, Gardner M, Fletcher C, Rickards E, Choo-Kang E. Effect of aqueous neem (Azadirachta indica) extract on testosterone and other blood constituents in male rats. A pilot study. West Indian Med J 1994;43:71-4. |
|24.||Mbah AU, Udeinya IJ, Shu EN, Chijioke CP, Nubila T, Udeinya F, et al. Fractionated neem leaf extract is safe and increases CD4+ cell levels in HIV/AIDS patients. Am J Ther 2007;14:369-74. |
|25.||O'Farrill-Santoscoy F, O'Farrill-Cadena M, Fragoso-Morales LE. Evaluation of treatment ofiron deficiency anemia in pregnancy. Ginecol Obstet Mex 2013;81:377-81. |
|26.||Townsley DM. Hematologic complications of pregnancy. Semin Hematol 2013;50:222-31. |
|27.||Hanieh S, Ha TT, Simpson JA, Casey GJ, Khuong NC, Thoang DD, et al. The effect of intermittent antenatal iron supplementation on maternal and infant outcomes in rural Viet Nam: A cluster randomised trial. PLOS Med 2013;10:e1001470. |
|28.||Raja SB, Murali MR, Kumar NK, Devaraj SN. Isolation and partial characterisation of a novel lectin from aegle marmelos fruit and its effect on adherence and invasion of shigellae to HT29 cells. PloS One 2011;6:e16231. |
|29.||Sen P, Mediratta PK, Ray A. Effects of Azadirachta indica A Juss on some biochemical, immunological and viscera parameters in normal and stressed rats. Indian J Exp Biol 1992;30:1170-5. |
|30.||Ray A, Banerjee BD, Sen P. Modulation of humoral and cell-mediated immune responses by Azadirachta indica (Neem) in mice. Indian J Exp Biol 1996;34:698-701. |
|31.||Shailey S, Basir SF. Strengthening of antioxidant defense by Azadirachta indica in alloxan diabetic rat tissues. J Ayurveda Integr Med 2012;3:130-5. |
|32.||Atangwho IJ, Ebong PE, Eyong EU, Asmawi MZ, Ahmad M. Synegistic antidiabeticactivity of Vernonia amygdalina and Azadirachta indica: Biochemical effects and possible mechanism. J Ethnopharmacol 2012;141:878-87. |
|33.||Akhtar N, Khan BA, Majid A, Khan HM, Mahmood T, Gulfishan, et al. Pharmaceutical and Biopharmaceutical evaluation of extracts from different plant parts of indigenous origin for their hypoglycaemic responses in rabbits. Acta Pol Pharm 2011;68:919-25. |
|34.||Jelodar GA, Maleki M, Motadayen MH, Sirus S. Effect of fenugreek, onion and garlic on blood glucose and histopathology of pancreas of alloxan-induced diabetic rats. Indian J Med Sci 2005;59:64-9. |
|35.||Chattopadhyay RR, Chattopadhyay RN, Nandy AK, Podder G, Maitra SK. Preliminary report on antihyperglycemic effect of a fraction of fresh leaves of Azadirachta indica (Beng. Neem). Bull Calcutta School Trop Med 1987;35:29-35. |
[Table 1], [Table 2], [Table 3]
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