Local Blend of Plant Improves Blood Parameters In Anaemia-induced Albino Wistar Rats.

Kevin E. Aghatise

Department of Medical Laboratory Science, Igbinedion University Okada

Ivie Osula and Anslem O. Ajugwo

Department of Medical Laboratory Science, Madonna University Nigeria

All Correspondences to: Kevin E. Aghatise, Department of Medical Laboratory Science, Igbinedion University Okada

ABSTRACT

Anaemia constitutes a serious health problem in many tropical countries including Nigeria because of the prevalence of malaria and other parasitic infections which possibly leads to decrease of hemoglobin. Telfairia occidentalis (fluted pumpkin), Vernonia amygdalina (bitter leaf) and Gongrone malatifolium (utazi leaf) have been reported to be very good in building the constituents of the blood and also replacing them. This study was designed to investigate the effect of fluted pumpkin, bitter leaf and utazi leaf extracts on haematolgical parameters of anaemia induced albino wistar rats. Fifteen Wistar albino rats with weight range from (122-190g) obtained from the animal house of Madonna University, Elele Nigeria were separated into three groups of five rats each for the study. Rats in group B and C were injected 400mg/kg 2,4 dinitrophenylhydrazine for five days to induce anaemia, while rats in group A were fed with grower’s mash and water throughout the experiment. All analysis were done using the standard methods. The result of this study show that, there was a significant (p<0.05) difference when the PCV was compared among the Groups, the control groups had 41.0+2.65% anemia induced group had 25.3+6.8% and the treatment group recorded 33.6+4.9%. The anemia induce group recorded a reduced PCV but was raised in the treatment group and this was statistically significant (p<0.05). There was also a significant difference when the Haemoglobin concentration was compared among the groups, the control group had 14.1+0.7g/dl, anaemia-induced group has 8.3+1.9g/dl and the treatment group had 10.+2.15g/dl. There was no significant difference (p>0.05) in all other parameters estimated. The result of this research indicate that aqueous leaf extract of fluted pumpkin, bitter leaf and utazi leaf could elevate the packed cell volume, red blood cells and hemoglobin concentration in anaemia-induced rats. It can be concluded that aqueous leaf extract of the combination of fluted pumpkin, bitter leaf and utazi leaf are a potential blood booster.

Keywords: anaemia-induced, blood booster, haematological, dinitrophenylhydrazine.

INTRODUCTION

The use of herbal products for medicinal benefits has an important role in nearly every culture on earth. Herbal medicine was practiced by ancient people of Africa, Asia, Europe and the Americas (Abosi and Raseroka, 2003). Over 50% of all modern clinical drugs are of natural product origin and natural products play an important role in drug development programs of the pharmaceutical industry. The consumption of a variety of local herbs and vegetables by man is believed to contribute significantly to the improvement of human health, in terms of prevention, or cure of disease because plants have long served as a useful and rational source of therapeutic agents (babalolaet al., 2003). Regular consumption of plant foods are associated with numerous health benefits rooted in their various physiological effects as a result of their physiochemical and nutritional constituent (Hunter and Fletcher, 2002).

Green leafy vegetables are particularly important in

promoting health because of their rich sources of nutrients (Gupta and Prakash, 2009). Telfairia occidentalis (fluted pumpkin) is a common tropical green leafy vegetables native to many African countries especially Eastern Nigeria (Burkett, 1968). It thrives in humid climate and well drained soils and is usually cultivated in garden and family farms around homes. Telfairia occidentalis is traditionally used by an estimated 30 to 35 million people indigenous people in Nigeria, including the Efik, Ibibio, and Urhobo (Akoroda, 1990). However, it is predominantly used by the Igbo tribe, who continue to cultivate it for food sources and traditional medicines (Okoli and Mgbeogu, 1983). A reoccurring subject in the Igbo’s folklore, the fluted gourd is noted to have healing properties and was used as a blood tonic, to be administered to the weak or ill (Akoroda, 1990). Bitter leaf is a medicinal plant of the family Asterceae. It is a small perennial shrub that grows in tropical Africa. Vernonia amygdalina is commonly called bitter leaf because of its bitter taste and it can be propagated either by cutting or seedling. Today, the plant is widely known throughout the continent and nearly 85% of Nigerians cultivates the plant due to its nutritional and medicinal values. It is locally called ‘ewuro; in Yoruba land (Abosi and Raseroka, 2003). The leaves of the plant may be consumed either as a vegetable (macerated leaves in soup) or aqueous extract as tonic for the treatment of various illnesses (Abosi and Reseroka, 2003).

Gongrone malatifolium also called utazi belongs to class of medicinal plants that are beneficial in preventing and treating certain disease and ailments that are detrimental to human health. Gongrone malatifolium leaf, which can be chewed infused or used for cooking is mainly used in the western part of America for nutritional and medicinal reason. Pharmacological studies suggest that utazi has both analgesic, antimicrobial, antibacterial, anti ulcer and antioxidant properties (Eguyoni et al., 2009). Utazi plant is highly medicinal in nature, which suggest why its health benefits cannot be overemphasized. Researchers agree that the fundamental ingredients used for medical purposes are stored in the various parts of the utazi plants such as leaves, fruits, roots etc (Eguyoni et al., 2009). The vital medicinal ingredients stored in the various parts of the utazi leaf can be extracted either through blending the fresh leaves, chewing the utazi seed, leaves or fruits, infusing either the dry or fresh utazi leaves and decoction. However these various methods of preparing and using utazi leaves for medical purpose mainly depends on the part of utazi plants where the active medicinal ingredient are present (Eguyoni et al., 2009).

Anaemia is decrease in the amount of red blood cells (RBCs) or hemoglobin in the blood. It can also be defined as a lowered ability of the blood to carry oxygen (Rodak and Bernadtte, 2007). When anaemia sets in slowly, the symptoms are often vague and may include: feeling tired, weakness, shortness of breath or a poor ability to exercise. Anaemia that comes on quickly often has greater symptoms, which may include; confusion, feeling like one is going to pass out, loss of consciousness, or increased thirst. Anaemia must be significant before a person becomes noticeably pale (Rodak and Bernadette, 2007).Use of plants has long been known to be used as local remedy for treatment of anaemia especially pumpkin leaf which is used in villages to treat anaemia especially when quality drugs cannot be accessed. This work was carried out to determine the effects of combination of herbal plants in anaemia-induced condition using haematological parameters.

Materials and Methods

Animal Model/Experimental design

Fifteen (15) Wistar strain albino rats with weight range of

(122 – 190g) obtained from the animal house, Madonna University Elele, Nigeria were used for the study. The rats were housed in wire meshed cage under standard conditions (temperature 25 – 29°C, 12 hours light and 12 hours darkness cycles) and fed with standard rat pelleted diet and water. They were housed in 3 meshed cages containing five

1. rats in each cage. The rats were made to acclimatize for 1 week before the experiment began. They were allowed to feed on standard feed and water freely throughout the period the experiment lasted.

Ethical Approval

The research was approved by the ethical committee of the institution. The standard, rules and regulations of use of animal for research purposes was strictly adhered to as approved by the committee.

Order of Placement

Group1: The rats in this group served as control and were

administered with feed and water only. Group2: The rats in this group served as test and were administered with feed, water and 1 ml of phenyl hydrazine for five days. Group3: The rats in this group served as treatment group and were administered with feed, water, 1 ml of phenyl hydrazine for five days and subsequently treated with 0.5 ml of a blend of the local plants for another five days. Preparation and Administration of the plant blend

Large quantities of fresh leaves of Telfairia occidentalis (fluted pumpkin), Vernonia amygdalina (bitter leaf) and Gongrone malatifolium (utazi leaf) were purchased from the Central Market Elele. Identification and authentication of the plants was done by Mr Eze of the Faculty of Science. The leaves were washed and dried air-dried for eight days until there was no sign of moisture and grinded into powder. The powder was dissolved in 100 ml of deionized water and administered to the treatment group 0.5 ml in the morning and evening for 5 days.

Sample Collection

2ml of blood sample was collected by ocular puncture from each of the animal model using capillary tube and was dispensed into commercially prepared concentrations of ethylene diamine tetra acetic acid containers.

Sample Analysis

Blood samples collected were analyzed within six hours of collection for Hb, PCV, RBC count, Total WBC and differential analysis using standard manual methods (Cheesebrough, 2000)

RESULTS

Mean±S.D of hematological parameters of the control group (A), test group (B) and treatment group ©

PARAMETERS	GROUP A	GROUP B	GROUP C	P-VALUE
PCV (%)	41.0+2.65	25.3+6.8	33.6+4.9	P<0.05
HB (g/dl)	14.1+0.7	8.3+1.9	10.5+2.15	P<0.05
RBC (X1012/l)	6.0+0.56	4.4+0.62	5.2+1.0	p>0.05
TWBC X109/1)	9.3+1.9	13.7+5.4	11.7+2.11	p>0.05
Platelet (x109/1)	272+24.0	261+39.5	302+39.2	p>0.05
NEUTROPHIL (%)	56.7+3.51	63.7+6.11	55.7+5.86	p>0.05
LYMPHOCYTE (%)	34+4.0	29.7+7.23	33.7+11.06	p>0.05
EOSINOPHIL (%)	6.0+1.0	4.0+1.0	4.3+1.53	p>0.05
MONOCYTE (%)	3.3+1.15	3.3+1.52	3.3+1.15	p>0.05

20

 

DISCUSSION

The consumption of a variety of local herbs and vegetables by man is believed to contribute significantly to the improvement of human health, in terms of prevention, and or cure of diseases because plants have long served as a useful and rational sources of therapeutic agents (Babalola et al., 2003). Regular consumption of plant foods are associated with numerous health benefits rooted in their various physiological effects as a result of their physiological and nutritional constituents (Hunter and Fletcher, 2002).

From the research work there was a significant (p<0.05) difference when the PCV was compared among the groups, the control group has 41.0±2.65%, anaemia induced group has 25.3±6.8% and the treatment group recorded 33.6±4.9%. The anaemia induced group recorded a reduced PCV but was raised in the treatment group and this was statistically significant (p<0.05). This definitely is attributable to the local blend of plants. Previous work have shown that pumpkin is capable of improving haematological parameters (Alada,2000; Adias et al., 2013).

There was also a significant difference when the Hb was compared among the group, the control group had 14.1±0.7g/dl, anaemic induced group had 8.3±1.9g/dl and the treatment group had 10.5±2.15g/dl, the anaemic group recorded a reduction 8.3+1.9/dl but was elevated in the group given the extract. The reason for the significant increase in these haematological parameters could be related to the chemical composition of the leaves used in this research work. The chemical composition of Telfairia occidentals includes protein, fat, carbohydrates, calcium, iron, vitamin A, thiamine and riboflavin(Tindal, 1968; Adisa et al., 2014). Most of these constituents have positive impact on blood production (Akube, 1980). For instance, iron is a well established haemopoetic factor and deficiency of if produces anaemia (Ganong, 1997). There was no significant (p>0.05) difference in the RBC, the control group had 6.0±0.56×1012/1, the anaemic group had 4.4±062×1012/1, while the treatment group had 5.2±1.0×1012/l. The extract increased the RBC count of the anaemia-induced group from 4.4+062×1012/1 to 5.2+1.0×1012/1 but this was not significant. For TWBC there was no significant (p>0.05) difference when the test groups B (13.7±5.4×109/1) was compared to the control group A (9.3±1×109/1) although the TWBC was increase in the anaemic group (13.7±5.4109/1) and reduced in the treatment group (11.7±2.11×109/1).

There was also no significant (p>0.05) difference in platelets count when the groups B (261±39.5×109/1) and C (302±39.2×109/1) was compared with control group

(272±24.0×109/1) although the platelets count decreased in the anaemic group and increased in the treatment group but this was not significant (p>0.05). No significant difference was seen in all the differential count parameters. In conclusion, the combination of the extract of Telfairia occidentalis (fluted pumpkin), Vernonia amygdalina (bitter leaf) and Gongrone malatifolium (utazi leaf) have been shown to improve blood parameters (PCV and haemoglobin concentration). This extract can be used in management of anaemia especially when the conventional blood builders are not readily available.

Conflict of interest

The Authors declare that we have no conflict of interest.

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