EFFECTS OF THE AQUEOUS AND ETHANOLIC EXTRACTS OF GNETUM AFRICANUM LEAVES ON BREWER’S YEAST INDUCED PYREXIA IN MALE WISTAR RATS

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ABSTRACT 
The need for an alternative method of managing high body temperature has become necessary due to the toxicity of antipyretic drugs on various organs in the body when taken over a long period of time. Gnetum africanum, a wild liana found in Nigeria, Central Africa, Asia and South America, is believed to possess some phytochemical constituents that can help in the management of fever. This study was carried out to determine the effects of aqueous and ethanolic leaves extracts of Gnetum africanum on yeast-induced pyrexia in male wistar rats. 50 acclimatized male wistar rats weighing 120g-200g were used for the study. After preparation of the extracts, A dark-green residue of the aqueous extract with a yield of 5.85g was obtained while the ethanolic extract obtained had a yield of 14.94g. The residues were reconstituted such that 1g of each extract was dissolved in 10mls of the different solvents to make up the concentration of the stock solution. Pyrexia was induced in all the experimental animals by subcutaneous injection of the brewer’s yeast suspension, their rectal temperatures were then measured and recorded after 24hrs. About 40 rats were selected for the study as they showed an increase in temperature up to 0.5oc. The animals were randomly assigned into 8 groups of 5 rats. Group 1 served as the control group and received 5ml/kg of 5% tween 80 solution. Group 2 received 500mg/kg of the reference drug, Paracetamol orally. Groups 3, 4 and 5 received 125, 250 and 500mg/kg dosages of the aqueous extract orally while Groups 6, 7 and 8 also received the same dosages of the ethanolic extract orally. After administration of the treatments/extracts, the temperatures of the animals were measured and recorded at 30mins, 60mins, 90mins and 120mins respectively. The results of the study showed that the different dosages of the ethanolic extract were able to reduce temperature significantly (p<0 .05="" 500mg="" a="" able="" an="" and="" animals="" aqueous="" at="" be="" both="" but="" compared="" could="" dosage="" dosages="" drug="" earlier="" early="" especially="" ethanolic="" extract="" findings="" higher="" i="" in="" interval.="" interval="" kg="" later="" leaves="" of="" p="" paracetamol="" post-treatment="" pyretic="" rats="" reduce="" reduced="" reference="" seen="" significantly="" similar="" suggest="" temperature="" that="" the="" therefore="" time="" to="" was="" were="" whereas="" which="" with="">Gnetum africanum
plant possess antipyretic properties which was exhibited in a dose-dependent manner, however, the ethanolic extract demonstrated more potency compared with the aqueous extract.

TABLE OF CONTENTS

Title Page
Table of Contents
Abstract

CHAPTER ONE:  INTRODUCTION
1.0   Introduction
1.1    Justification
1.2    Aim of the Study
1.3    Objectives of the Study

CHAPTER TWO: LITERATURE REVIEW
2.0    Normal body temperature
2.1    Regulation of body temperature
2.2    Fever, Its Causes and Symptoms
2.3    Effects of fever on the body
2.4    Methods of Inducing fever in laboratory Animals
2.5    Composition of Brewer’s yeast
2.6    Non-Pharmacological methods used to manage pyrexia
2.7    Pharmacological methods used to manage pyrexia
2.8    The use of plants in the management of pyrexia
2.9    Diagram of Gnetum africanum plant

CHAPTER THREE: MATERIALS AND METHODS
3.1    Collection of Plant Sample, Identification and Authentication
3.2    Preparation of extracts of Gnetum africanum
3.3    Experimental Animals
3.4   Oral toxicity study
3.5   Brewer’s yeast, drugs and other reagents
3.6   Phytochemical analysis
3.6.1 Qualitative test
3.6.1a Preparation of Wagner’s reagent
3.6.1b Preparation of Mayer’s reagent
3.6.1c Test for Alkaloids
3.6.1d  Test for Saponins
3.6.1e  Test for Flavonoids
3.6.1f    Test for Steroids
3.6.1g    Test for Tannins
3.6.1h    Test for Phenols
3.6.1i     Test for Glycosides
3.6.2      Quantitative Analysis
3.6.2a    Determination of total phenol
3.6.2b   Alkaloid determination
3.6.2c    Flavonoid determination
3.6.2d   Saponin determination
3.6.2e   Tannin determination
3.6.2f    Glycoside determination
3.7        Oral toxicity study
3.8       Induction of pyrexia
3.9      Animal grouping and administration of treatments/extracts

CHAPTER FOUR:  RESULTS
4.0    Statistical Analysis
4.1 Table showing qualitative analysis of extracts of Gnetum africanum
4.2 Table showing quantitative analysis of extracts of Gnetum africanum
4.3 Table showing effects of aqueous extracts of Gnetum africanum leaves on pyrexia
4.4 Table showing effects of ethanolic extracts of Gnetumafricanum leaves on pyrexia
4.5 Level of reduction in temperature at the different post-treatment time intervals
4.5.1 Line graph at 30mins post- treatment time interval
4.5.2 Line graph at 60mins post-treatment time interval
4.5.3 Line graph at 90mins post-treatment time interval
4.5.4 Line graph at 120mins post-treatment time interval

CHAPTER FIVE:  DISCUSSION
5.0 Discussion
5.1 Conclusion
5.2 Recommendation
References


CHAPTER 1
1.0 INTRODUCTION
The body usually maintains a set temperature as humans are homeotherms with body organs that function efficiently at a relatively constant temperature that ranges between 36.5oc-37.5oc, though there could be slight variations with this temperature range due to several environmental and metabolic factors. The normal body temperature varies among individuals depending on gender, recent activity or level of activity, food and fluid consumption, menstrual variations in females, the time of the day, as body temperature is usually lowest just before dawn, it increases slightly in the morning in the morning till noon and higher temperatures occur from afternoon till early evening. Normal body temperature is regulated by temperature-regulating centres which are found in the pre-optic area in the anterior hypothalamus which try to strike a balance between heat loss and heat gain. The temperature regulatory system is controlled by a nervous feedback mechanism such that mechanisms such as vasodilatation, sweating etc are used to restore high body temperature to normal, conversely, a low body temperature is restored back to normal through vasoconstriction, piloerection, shivering etc (Nasir, 2011).
A temporary increase in body temperature is used by the body to combat infections, however, when there is a sustained increase in body temperature above the normal physiologic range, Fever or Pyrexia occurs (Chattopadhyay et al, 2005, Spacer and Breder, 1994) and this requires medical attention. This condition is triggered by pyrogens, infectious agents and other environmental and metabolic factors which cause a rise in the set-point of the hypothalamic thermostat and it can lead to reduction in the volumes of fluid in the body, increased breakdown of protein and other nitrogenous substances, weakness and other severe conditions in the body.                           
Antipyretic drugs which are also called Non-steroidal anti-inflammatory drugs have been used for a long time in the management of fever and these include Aspirin, Ibuprofen, Acetaminophen(Paracetamol) etc. Their mechanism of action is by inhibition of prostaglandin synthetase thereby inhibiting the production of prostaglandin within the hypothalamus or by suppressing increase in Interleukin-1 production subsequent to interferon production (Clark and Cumby, 1999, Kurokawa et al, 1996).
The use of plants as an alternative form of therapy in primary healthcare of individuals and communities in most developing countries has increased in recent times as most plants and their derivatives have been discovered to be highly effective in the treatment of various diseases.
Gnetum africanum, an evergreen climbing vine, which belongs to the family of Gnetaceae, is a plant that has several nutritional and medicinal benefits. The presence of flavonoids and other phytochemical constituents in this plant is likely to enhance its ability to reduce high body temperature by suppressing production of prostaglandin through inhibition of prostaglandin synthase. The ability of the extracts of this plant to reduce pyrexia in male wistar rats will contribute to existing knowledge about its medicinal benefits.

1.1 JUSTIFICATION FOR THE STUDY
Fever is a common symptom associated with most disease conditions in the body and a high level of fever can lead to serious conditions as a result of which proper management of this condition is necessary to prevent such complications(Spacer and Breder, 1994). Antipyretic drugs have been in use and are still in use for the management of fever, however, excessive consumption of these drugs has several side effects on the body. As a result of the toxicity of these drugs to various organs of the body when taken for a long period of time, there is a growing need for a safer and alternative form of therapy. Plant extracts might therefore provide the needed alternative method of managing this condition of fever. Flavonoids, which are one of the phytochemical constituents present in Gnetum africanum plant, are water soluble polyphenolic molecules found in most plant material. They have different effects in the body such as anti-allergic, anti-oxidant, anti-inflammatory and anti-viral effects (Cushnie and lamb, 2005).Several flavonoids are also known for their antipyretic effect through the inhibition of prostaglandin synthase (COX-2) production and their presence in Gnetum africanum leaves is hypothesized to help in reducing fever when it is consumed by individuals as a form of herbal therapy.

1.2 AIM OF THE STUDY
This study was aimed at investigating the pyretic activity of aqueous and ethanolic leaf extracts of Gnetum africanum on male wistar rats.

1.3   OBJECTIVES OF THE STUDY
To determine the effect of the aqueous extracts of Gnetum africanum leaves on the temperature of wistar rats induced with pyrexia using brewer’s yeast.
To check the effect of the ethanolic extracts of Gnetum africanum leaves on the temperature of wistar rats induced with pyrexia using brewer’s yeast.

To compare the effects of both the aqueous and ethanolic extracts of Gnetum africanum on the temperature of wistar rats induced with pyrexia using brewer’s yeast.....

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