ABSTRACT
The high prevalence, complications and cost of conventional
drugs in the management diabetes necessitated the search for alternative
treatment. As a result, this study seeks to evaluate the composition,
anti-diabetic potential toxicity and tissue-protective effects of both the
water and methanolic extracts of Persea americana (avocado pear) seed on
alloxan-induced diabetic albino rats were investigated. This study was
conceived and designed based on information on the local use of the seed in
diabetes treatment. Proximate and anti-nutritional constituents of the seed
were determined and 100g of the sample was extracted with 1000ml of both water
and methanol using the maceration method. The extracts were evaporated to
dryness using a rotary evaporator and stored at 4oC
until use. The effects of different doses (200mg/kg.b.wt., 300/kg.b.wt.) of
both water and methanolic extracts of P. americana seed on alloxan-induced
diabetic albino rats were compared with those of a reference drug, insulin. The
glucose level and weight of the rats were measured weekly for 21 days. The
liver function tests and the histopathologies of the liver, and kidneys, were
investigated. Results of the proximate investigation shows that the seed is
rich in carbohydrate (49.03± 0.02 g/100g), lipid (17.90±
0.14 g/100g), protein (15.55± 0.36 g/100g) moisture (15.10±
0.14 g/100g) and ash (2.26±0.23 g/100g). Anti nutritional components such as
total oxalate (14.98±0.03 mg/100g), tannin (6.98±0.04 mg/100g) and phytic acid
(3.18±0.16 mg/1 00g). Results also showed that both the water and methanolic
extracts exhibited significant anti-diabetic effects on the experimental rats.
However, the methanolic extracts showed a better anti diabetic effect than the
water extracts. The extracts showed no significant effects on the liver
function parameters (bilirubin, conjugate bilirubin, AST, ALP and ALT) compared
with the normal control but rather reversed the histopathological damage that
occurred in alloxan-induced albino diabetic rats. In conclusion, the present
study provides a pharmacological basis for the traditional use of P. americana
seeds extracts in the management of Diabetes mellitus. It seems P. americana
seed contains substantial amount of nutrients that could warrant its
utilization in animal feed or food. However, further studies are required to
indentify the active ingredient responsible for the anti-diabetic properties of
the seed extract.
TABLE OF CONTENT
Title Page
Certification
Dedication
Acknowledgement
Abstract
Table of Contents
List of Tables
List of Figures
CHAPTER ONE
1.0 Introduction
1.1 Background
1.2 Global Burden
of Diabetes
1.3 Statement of
Problem
1.4 Aims and
Objectives of the Study
CHAPTER TWO
2.0 Literature
Review
2.1 Diabetes
Mellitus
2.1.1 Types of
Diabetes Mellitus
2.1.2 Prevention /
Treatment of Diabetes Mellitus
2.2 Medicinal
Plants
2.3 Evaluation of
Drug Toxicity
2.3.1 Liver Function
2.3.1.1 Total Bilurubin
2.3.1.2 Conjugated
Bilurubin
2.3.1.3 Aspartate
Transaminase (AST)
2.3.1.4 Alanine
Transaminase (ALT)
2.3.1.5 Alkaline
Phosphatase (ALP)
2.3.2 Histology of
Liver and Kidney
CHAPTER THREE
3.0 Materials and
Methods
3.1 Experimental
Design
3.2 Chemicals and
Reagents
3.3 Sample
Collection and Preparation
3.4 Proximate
Analysis
3.4.1 Protein Content
3.4.2 Crude Fat
Content
3.4.3 Moisture
Content
3.4.4 Ash Content
Measure
3.4.5 Carbohydrate
Content
3.5 Anti-nutritional
Components
3.5.1 Tannin Content
Determination
3.5.2 Oxalate
Determination
3.5.3 Phytic Acid
3.6 Animal
Experiment
3.6.1 Induction of
Diabetes
3.6.2 Administration
of Extracts
3.6.3 Blood Sample
Collection
3.6.4 Liver Function
Tests
3.6.4.1 Total Bilirubin
Test
3.6.4.2 Conjugated
Bilirubin Test
3.6.4.3 Alanine
Aminotransferase (ALT) Test
3.6.4.3 Aspartate
Aminotransferase (AST) Test
3.6.4.5 Alkaline
Phosphatase (ALP) Test
3.6.5 Histological
Study
3.6.5.1 Histology Sample
Preparation
3.7 Statistical
Analysis of Data
CHAPTER FOUR
4.0 Results
4.1 Proximate
Analysis
4.2 Anti-Nutritional
Analysis
4.3 Animal
Experiment Results
4.3.1 Glucose
Analysis Results
4.3.2. Results of Body
Weight Analysis
4.3.3. Liver Function
Test Results
4.3.4 Histology
Results
CHAPTER FIVE
5.0 Discussion
5.1 Proximate and
Anti-nutrients
5.2 Effects of
Extracts on Body Weight
5.3 Effects of
Seed Extracts on Blood Glucose
5.4 Effects of
Seed Extracts on Liver Function Parameters
5.5 Histological
Effects of Extracts on the Kidney and Liver
5.6 Conclusion
References
Appendix
.
LIST OF TABLES
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Tables
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Pages
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Table 1:
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Result of the proximate analysis
of P. american seed (g/100g)
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Table 2:
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Anti-nutritional constituents of P.
americana seed (mg/100g) -
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Table 3:
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Effect of Extracts on Blood
glucose levels (mmol/L) of Normal
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and Diabetic rats
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Table 4:
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Effect of Extracts on Mean
Bodyweight (g) of Normal and
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Diabetic rats
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Table 5:
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Effect of Extracts on Mean Liver
Function Parameters of Normal
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and Diabetic rats
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LIST OF
FIGURES
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Figures
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Pages
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Fig.1:
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Section of liver tissue of normal
rat
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Fig. 2:
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Cortical section of kidney tissue
of normal rat
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Fig. 3:
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Section of liver tissue of
untreated diabetic rat following liver damage
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by alloxan.
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Fig. 4:
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Section of kidney tissue of
untreated diabetic rat following kidney
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damage by alloxan.
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Fig. 5:
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Section of
liver tissue of rat treated with 1unit of 40µ/50g b.w./day dose
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of insulin following liver damage
by insulin. -
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Fig. 6:
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Section of kidney tissue of rat treated
with 1unit of 40µ/50g b.w./day
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dose of insulin following kidney
damage induced by alloxan.
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Fig. 7:
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Section of liver tissue of rat
treated with 200mg/kg of water extract of
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avocado seed following liver
damage induced by alloxan.
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Fig. 8:
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A section of kidney tissue of rat
treated with 200mg/kg of water extract
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of avocado seed following kidney
damage by alloxan.
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Fig. 9:
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Section of rat
liver treated with 300mg/kg water extract of avocado seed | |
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following liver damage by alloxan.
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Fig. 10:
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Section of rat kidney tissue
treated with 300mg/kg water extract of
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avocado seed following kidney
damage by alloxan. -
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Fig. 11:
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Section of liver tissue of rat
treated with 200mg/kg dose of methanolic
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extract of avocado seed following
liver damage by alloxan.
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Fig. 12:
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Section of medulla of kidney
tissue of rat treated with 200mg/kg dose of
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methanolic extract of avocado seed
following kidney damage by alloxan.
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Fig. 13:
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Section of liver tissue of rat
treated with 300mg/kg dose of methanolic
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extract of avocado seed extract
following liver damage by alloxan. -
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Fig. 14:
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Medullary section of kidney of rat
treated with 300mg/kg dose of
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methanolic extract of avocado seed
extract following kidney damage by
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alloxan.
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Fig. 15:
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Mean Body weight curve of normal
Rats
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Fig. 16:
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Mean Body weight curve of diabetic
untreated Rats -
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Fig. 17:
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Mean Body weight of insulin
treated diabetic Rats
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Fig. 18:
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Effect of 200mg/b.wt. water
extract treatment on diabetic Rats
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Fig. 19:
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Effect of 300mg/b.wt. water
extract treatment on diabetic Rats
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Fig. 20:
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Effect of 200mg/b.wt. methanolic
extract treatment on diabetic Rats
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Fig. 21:
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Effect of 300mg/b.wt. methanolic
extract treatment on diabetic Rats
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Fig. 22:
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Effect of Treatments on Mean
Bodyweight at Various Intervals.
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Fig. 23:
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Mean blood glucose levels of all
the groups before alloxan induction.
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Fig. 24:
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Mean blood glucose levels for diabetic
rats treated with insulin, water
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and methanol avocado seed extracts
at day 1 of treatment.
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Fig. 25:
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Mean blood glucose levels for
diabetic rats treated with insulin, water
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and methanol avocado seed extracts
at day 1 of treatment.
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Fig. 26:
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Mean blood glucose levels for
diabetic rats treated with insulin, water
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and methanol avocado seed extracts
at day 14 of treatment. -
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Fig. 27:
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Mean blood glucose levels for
diabetic rats treated with insulin, water
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and methanol avocado seed extracts
at day 21 of treatment. -
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Fig. 28:
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Summary of the Blood Glucose level
for diabetic rats treated with insulin,
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water and methanol avocado seed
extract. -
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Fig. 29:
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Liver function parameters of
alloxan induced diabetic rats treated with
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insulin, methanol and water
extracts of P. Americana seed. -
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Fig. 30:
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Mean Total Bilirubin concentration
of diabetic rats treated with insulin
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and varied doses of water and
methanol extracts of avocado seed.
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Fig. 31:
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Mean Conjugate Bilirubin
concentration of diabetic rats treated with
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insulin, and varied doses of water
and methanol extracts of avocado seed.
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Fig. 32:
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Mean Aspartate Transaminase
concentration of diabetic rats treated with
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insulin, and varied doses of water
and methanol extracts of avocado
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seed.
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Fig. 33:
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Mean Alanine Transaminase
concentration of diabetic rats treated with
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insulin, and varied doses of water
and methanol extracts of avocado
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seed.
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Fig. 34:
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Mean Alkaline Phosphatase
concentration of diabetic rats treated with
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insulin, and varied doses of water
and methanol extracts of avocado
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seed.
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CHAPTER ONE
1.0
INTRODUCTION
1.1
BACKGROUND
During the past decade, the traditional systems have gained
importance in the field of medicine. The World Health Organization estimates
that 4 billion people, 80% of the world population (WHO, 2002), presently use
herbal medicine for some aspect of primary health care (Orisataoki and
Oguntibeju, 2010). Eighty percent (80%) of African populations use some form of
traditional herbal medicine (WHO, 2002.) and the worldwide annual market for
these products approaches US$ 60 billion (Willcox and Bodeker, 2004). Herbal
medicine is a major component in all indigenous people's traditional medicine.
Medicinal plants have continued to attract attention in global search for
effective methods of using plants parts (e.g. seeds, leaves, stems, roots,
barks etc) for the treatment of many diseases affecting humans (Sofowora,
2008). This is as a result of the continuous need for less expensive means of
disease control.
Medicinal plants are plants which can be used for
therapeutic purposes or which are precursors for the synthesis of useful drugs
(Sofowora, 2008). Many important drugs used in healthcare today are directly
derived from plants due to its bioactive constituents such as; alkaloids,
tannins, steroids, etc. Examples include L-Dopa derived from Mucuna spp
used for anti-parkinsonism, another is Caffeine, a CNS stimulant derived from Camellia
sinensis and Quinine from Cinchona ledgeriana and used for
Antimalarial, antipyretic.
As a matter of fact, well into the 20th
century, much of the pharmacopoeia of scientific medicine was derived from the
herbal lore of native people. Many drugs commonly used today are of herbal
origin. Undisputedly, the history of herbology is inextricably intertwined with
that of modern medicine. Many drugs listed as conventional drugs were
originally derived from plants.
Salicyclic acid, a precursor of aspirin, was originally
derived from white willow bark and the meadowsweet plant (Zand, et al.,
2003).
Plants are very unique as their existence is very essential
for the sustenance of the rest of the food chain. Based on the observations
made through successive generations, superstition as well as traditional
medicinal folklore, man has found and has been using herbs, barks, fruits,
leaves, seeds, roots and stems of different plants of various climatic regions
for therapeutic purposes (Sofowora, 2008).
1.2
GLOBAL BURDEN OF DIABETES
Diabetes mellitus is a metabolic disease caused by a
deficiency in the secretion or action of insulin (Nelson and Cox, 2005). This
disorder is characterized by major symptoms as; polyuria (frequent and abundant
urine), glycosuria (presence of glucose in urine) and hyperglycemia (glucose
rate on an empty stomach higher than 1.2g/l in plasma blood and confirmed in at
least two occasion) (N’guessan et al., 2009). Basically, there are two
major clinical classes of diabetes
.The type 1 diabetes or insulin dependent diabetes mellitus
(IDDM) and type 2 diabetes or non insulin dependent diabetes mellitus (NIDDM)
also called insulin resistant diabetes. According to the International Diabetes
Federation (IDF) 2014 updates, out of the world seven billion population,
387million people, aged 20–79 years wor ldwide are diabetic, (IDF, 2014) giving
a comparative prevalence of 8.3%, while 46.3% cases are undiagnosed. In every 7
seconds, a person dies of diabetes, 4.9 million deaths in 2014. Seventy seven
percent (77%) of people with diabetes live in low and middle income countries.
Africa has recorded cases of 2,150,274 (5.05%) diabetic patients with over
13million undiagnosed cases. In Nigeria, there are estimated 374,651 diabetic
cases, with another 172,339 undiagnosed cases. These figures account for about 4.64% Nigerian adults between ages 20-79 living with
diabetes (IDF, 2014). In 2014, about 105,090 Nigerians died as a result of
diabetes (IDF, 2014). An average diabetic Nigerian spent about 43527.16 naira
(US $178.39) in 2014 due to diabetes treatment (IDF, 2014). With this alarming
prevalence rate, diabetes mellitus poses a major challenge globally and
accounts for a number of disabilities and deaths globally.
Currently, diabetes therapy is based on the use of
hypoglycemic drugs (sulfonamides, biguanides, and insulin), on hygieno-diet
measures, exercise, and requires a lifelong treatment. With the level of
poverty in developing nations like Nigeria, the need for a better and cheaper
medication cannot be over emphasized. Traditional medicine has always provided
a cheaper and time trusted alternative for the treatment and management of
various diseases over time.
1.3
STATEMENT OF PROBLEM
There is an urgent
need to provide alternative and cheaper means for the management and treatment
of diabetes.
1.4
AIMS AND OBJECTIVES OF THE STUDY
This study is aimed
at the evaluation of the anti-diabetic effects of water and mMETHANOLic
extracts of Persea americana (avocado) seed on alloxan induced diabetic
rats.
The
specific objectives of the study are:
1. To determine nutritional and some
anti-nutritional constituents of seed extracts of the plant Persea
americana.
2. To evaluate the anti-diabetic properties of seed extracts of
P. americana in diabetic rats
3. To evaluate the effect of P.
americana seed extract on liver function enzymes.
4. To investigate the histopathological
effects of P. americana on both the kidney and liver of diabetic rats.
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