THE ROLE OF n-BUTANOL LEAF fraction OF Telfairia occidentalis ON SOME HAEMATOLOGICAL PARAMETERS IN PHENYL-HYDRAZINE INDUCED ANEMIA IN WISTAR RATS Â
 ABSTRACT
Anemia is a serious health problem especially in developing countries. In the tropics, rural dwellers had resort to herbal treatment in some cases of anemia. The main aim of this work was to ascertain the role of n-butanol leaf fraction of Telfairia Occidentalis on some haematological parameters in phenyl hydrazine induced anemia in wistar rats. Thirty (30) adult Wistar rats were used for this study. All animals were assayed for haematological parameters 1day before the onset of the experiment. The animals were grouped into five groups of five rats each (n =5). Anemia was induced in the Wistar rats by intraperitoneal injection of Phenyl hydrazine Hydrochloride 50mg/kg w/w in DSMO once daily for 3 days. Group i. serves as the negative control group, received 50mg/kg of Phenylhydrazine Hydrochloride and 1ml/kg normal saline (untreated) group ii. serve as Positive control group received Vitamin B12 0.4ml/kg (standard haematinic) intramuscularly,while group iii, iv. and v. received 100mg/kg, 200mg/kg and 300mg/kg body weight of n-butanol leaf fraction of Telfairia occidentalis for 2 weeks respectively. Asessment of RBC and WBC was carried out using the newly improved Neubauer counting chamber, PCV using the microhaematocrit reader, Hb using the AO-Hb meter and Bilirubin concentration using colorimetric estimation for the serum bilirubin. There was a significant (P<0.05) increase in the level of RBC, PCV, HB and Bilirubin concentration after treatment with the fraction as compared to the control groups respectively. RBC (5.30±0.0217 x106/ʯL as compared to 4.78±0.17x106/ʯL), PCV (50.60±0.51% as compared to 37.08±0.37%), HB (16.84±0.18 g/dl as compared to 12.56±0.13 g/dl), WBC (6.10±0.15 x109/L as compared to 6.17±0.88x109/L) and Bilirubin (18.16±0.08a ʯmol/L as compared to 15.54±0.21 ʯmol/L). However, the significant increase obtained from the results of RBC ,PCV and Bilurubin was not dose dependent while that of HB and WBC was dose dependent. In conclusion, Intraperitoneal administration of 50mg/kg Phenylhydrazine hydrochloride for 3days decreases the blood parameters below the pre-anemic level (1day before induction) due to the production of reactive oxygen species. Phytochemical screening of this leaf fraction indicates the presence of flavonoids, saponin secoiridoid glycosides and alkaloids, these natural antioxidants could be responsible for reversing the damaging effect of PHZ and thus playing a modulatory role and also maintaining the integrity of the RBC.
TABLE OF CONTENTS
Title page i
Certification ii
Declaration iii
Acknowledgement iv
Abstract v
Table of contents vii
Lists of Figures, Tables, Plates and Appendices ix
Abbreviations Definitions, Glossary and Symbols xii
Chapter One
Introduction1
Statement of research problem3
Justification4
Hypothesis5
Specific aim and objective5
Chapter Two
Literature Review7
Vitamin B128
Phenyl hydrazine HCL10
Anemia among women13
Decrease haemoglobin-Oxygen affinity14
Redistribution of blood flow15
Increase Cardiac Output15
Classification of anemia17
Cytometric classification17
Erythrokinetic classification18
Reticulocyte counts18
Serum heptaglobin concentration19
Bone marrow biopsy20
Biochemical classification21
Macrocytic anemia22
Normocytic anemia22
Microcytic anemia24
Dimorphic anemia25
Heinz body25
Hyperanemia25
Grading of anemia25
Causes of anemia26
Increase destruction of Red blood cell27
Intrinsic and Extrinsic factors27
Mechanical trauma of Red cell28
Autoimmune haemolytic anemia28
2.61 Blood loss 29
Treatment of anemia30
Anemia and Cardiovascular system31
Anemia and oxygen delivery32
Anemia and haematocrit32
Anemia and cardio-respiratory adjust33
2.67 Anemia and cancer 35
Anemia and pregnancy36
Anemia and kidney37
Anemia and liver37
Portal Circulation and Anemia40
Impaired blood coagulation41
Aplastic anemia41
Alcohol, liver disease and anemia43
Chapter Three
3.0 Materials and Methods 45
Collection and identification of plant material45
Method of extraction45
Animals45
3 .2 Chemicals 46
3.3 Acute Toxicity studies 46
Experimental design46
Induction of anemia47
Animal groupings47
Sample collection47
Determination of haemoglobinconcentration48
Determination of the pack cell volume48
Biochemical assay49
Determinatoin of Bilirubin concentration49
3.6 Statistical analysis 49
Chapter Four
Results50
The role of n-butanol leaf fraction of T. Occidentalis on RBC50
The role of n-butanol leaf fraction of T. Occidentalis on WBC52
The role of n-butanol leaf fraction of T. Occidentalis on PCV54
The role of n-butanol leaf fraction of T. Occidentalis on HB58
The role of n-butanol leaf fraction of T Occidentalis on Bil conc60
Chapter Five
Discussion, Conclusion and Recommendation60
Discussion60
Conclusion62
Recommendations63
REFERENCE 64
CHAPTER ONE
INTRODUCTION
Anemia is a global public health problem affecting both developing and developed countries with major consequences for human health as well as social and economic development (WHO, 2005).
Anemia is a reduction from the normal quantity of circulating hemoglobin in the blood less than 13 g/dl for male and less than 12 g/dl for female adults (Okochi et al., 2003).  It occurs at all stages of the life cycle, but is more prevalent in pregnant women and young children (Adam et al., 2005).
Hemolytic anemia is a form of anemia due to hemolysis, the abnormal breakdown of Red
Blood Cells (RBCs), either in the blood vessels (intravascular hemolysis) or elsewhere in the human body (extravascular). It has numerous possible causes, ranging from relatively harmless to life-threatening (Telford et al., 2003). The general classification of hemolytic anemia is either inherited or acquired. Treatment depends on the cause and nature of the breakdown (Yamoto et al., 1998).
Anemia is the result of a wide variety of causes that can be isolated, but more often coexist. Globally, the most significant contributor to the onset of anemia is iron deficiency so that IDA and anemia are often used synonymously, and the prevalence of anemia has often been used as a proxy for IDA (De Maeyer, 1989). It is generally assumed that 50% of the cases of anemia are due to iron deficiency, but the proportion may vary among population groups and in different areas according to the local conditions (De Maeyer, 1989).
Anemia is an indicator of both poor nutrition and poor health. The most dramatic health effects of anemia is increased risk of maternal and child mortality which have been well documented (Irwin and Kirchner, 2001). In addition, anemia should be built into the primary health care system and existing programmes. These strategies should be tailored to local conditions, taking into account the specific etiology and prevalence of anemia in a given setting and population group (Benoist et al., 2008).
The importance of medicinal plants in traditional healthcare practices, providing clues to new areas of research and in biodiversity conservation is now well recognized. The use of traditional medicine and medicinal plants in most developing countries as a normative basis for the maintenance of good health has been widely observed (Ranjan et al., 2010). The search for new pharmacologically active agents obtained by screening natural sources such as microbial fermentations and plant extracts has led to the discovery of many clinically useful drugs that play a major role in the treatment of human diseases (Kumar et al., 2009). Telfairia occidentalis is a tropical vine grown in West Africa as a leaf vegetable for its edible seeds. Lately the popularity has spanned across other ethnic groups, thereby taking the medicinal and commercial value of the vegetable to a higher level. (Alada, 2000).
1.0.1 Nutritional Content
This blood-building vegetable is known to contain protein, carbohydrate, fat, calcium, iron, magnesium, potassium, calcium, and vitamin such as A, B2, B5, B12 and thiamine.(Harte et al., 1992).
Figure 1.1 Ugu Leaf (Telfairia Occidentalis) adopted from Journal of plant nutrition,
2011.
T. occidentalis leaves contain 30.5% crude protein, 3.0% crude lipid, 87.3% crude fibre, and 8.4% total ash. The leaves had low level of tannic acid (0.5mg/100 g DM) and oxalate (4 mg/100 g DM), but high level of phytic acid (12 mg/100 g DM) (Oboh,.2005).
STATEMENT OF RESEARCH PROBLEM
An estimated 30% of the world’s population is afflicted with anemia. Anemia in pregnancy is a leading cause of maternal and perinatal deaths in developing countries. In developing countries, anemia affects almost two thirds of the pregnant population. It is also estimated that anemia is responsible for as much as 20% of all maternal deaths in Sub-Saharan Africa (WHO, 2005).
Anemia in pregnancy in Nigeria is not different from other parts of sub-Saharan Africa. In the neighboring republic of Niger, Murray, et al., found that about 47% of breast-feeding women were iron deficient. Harrison, et al., (1989) showed that about 40% of pregnant women are anemic. Isah, et al., (1985) found a prevalence of anemia of 46% in non- pregnant and 52% of pregnant women respectively in Zaria. Iron deficiency was found in 54% and 25% of non-pregnant and pregnant women respectively by Isah, et al. Gwarzo, et al., (1994) in Kano found a prevalence of iron deficiency of 17% among pregnant women in Kano. With increase in the prevalence of anemia in developing countries such as Nigeria. It is important to assess the role of n-batanol fraction of Telfairia Occidentalis on anemia.
The leaves of Telfairia Occidentalis have been reported to possess anit-microbial, anti- fungal properties and also, anti-cholesterolemia (Oluwole, et al., 2003). The young leaves sliced and mixed with coconut water and salt are stored in a bottle and used for the treatment of convulsion in ethno medicine (Gbile, 2003) The leaf extract is useful in the management of cholesterolemia, liver problems and impaired defense immune systems (Eseyin, et al., 2005).
JUSTIFICATION
Although there are various drugs for the treatment of anemia, they are not accessible to many poor people especially those in the developing countries such as Nigeria. In addition, the rural populations in various parts of the world do not have adequate access to high quality drugs for the treatment of anemia, so they depend heavily on plants and herbal products for the treatment of diseases and anemia.
As a result of the fact that anemia is very common and the incidence is likely to increase in future, there is need to prevent it or seek for more cost-effective and better treatment strategies (Duff, 2008).
Available studies reveal that the leaf extract of Telfairia occidentalis can be used to treat Anemia, convulsion, artherosclerotic cardio vascular diseases, high blood pressure, hypocholesterolemia, arthritis, liver problems, and inflammatory conditions (Oyolu, 1978).
Most of the studies carried out use aqueous or crude extract of this plant on haematological indices (Ogbe, et al., 2010).There is need for the isolation and identification of the active compounds responsible for the anti-anemic activities.
In this study, we considered the role of n-butanol leaf fraction of Telfairia occidentalis on some haematological parameters in phenyl hydrazine induced anemia in Wistar rats.
HYPOTHESIS
The n-batanol leaf frction of Telfairia occidentalis possess anti-anemic properties.
AIM AND OBJECTIVES
To the role of n-butanol leaf fraction of Telfairia occidentalis on some haematological indices in phenylhydrazine-induce anemia in Wistar rats.
SPECIFIC OBJECTIVES
1. To ascertain the role of n-butanol leaf fraction of T. Occidentalis on PCV in phenylhydrazine induced anemia in Wistar rats.
2. To ascertain the role of n-butanol leaf fraction of T. occidentalis on RBC count in phenyl hydrazine induced anemia in Wistar rats.
3. To ascertain the role of n-butanol leaf fraction of T. occidentalis on WBC count in phenyl hydrazine induced anemia in Wistar rats.
4. To ascertain the role of n-butanol leaf fraction of T. occidentalis on HB concentration in phenyl hydrazine induced anemia in Wistar rats.
5. To ascertain the role of n-butanol leaf fraction of T occidentalis on Bilirubin concentration in phenyl hydrazine induced anemia in Wistar rats.
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