ABSTRACT
Diabetes mellitus (DM) is a metabolic disorder characterized by a loss of glucose homeostasis, due to disturbance of carbohydrate, fats, and protein metabolism, resulting from defects in insulin production and action or both. In Nigeria, the number of people suffering from diabetes mellitus is rapidly increasing. There are several oral hypoglycemic drugs used in the treatment of DM but are not devoid of adverse effect. However, natural products from plant source have proven to be effective and safe in the treatment of DM. An ethnobotanical survey conducted in Imo state (Isiala Mbano & Owerri) and Abia state (Ikwuano) in South Eastern Nigeria revealed nine medicinal plants frequently mentioned by the traditional healers in the treatment of diabetes mellitus. Nine medicinal plants namely; Chlorophora excelsa (root), Strophanthus hispisdus (root), Picralima nitida (seed), Persea americana (seed), Loranthus micranthus (leaf), Ceiba pentandra (leaf), Synsepalum dolficicum (leaf), Anthocleista djalonensis (leaf) and Anacardium occidentale (leaf). The phytochemical properties of these plants were estimated using standard TLC method while the antioxidant activities were done using 1, 1-diphenyl-2-picrylhydrazl (DPPH), 2, 21-azino-bis-3-ethylbenzothiazoline-6-sulfonic-acid (ABTS), and Ferric Reducing Antioxidant Power (FRAP).Some of the samples had (Ceiba pentandra and Anacadium occidentale) a robust antioxidant potential while some of the samples exhibited moderate antioxidant potential. The thin layer Chromatography (TLC) profiling revealed the presence of phenolics, flavonoids, alkaloids, saponins with yellow-brown coloured zones at daylight indicating the presence of alkaloid, yellow coloured zone indicating the presence of flavonoids, and blue coloured zone indicating the presence of phenols. The presence of these compounds in the plantsindicates the presence of phytochemical in the plant extracts in support of their antioxidant and biological activity. The α-amylase inhibition test indicated that some of the plant extracts (Chlorophora excelsa, Picralima nitida, Synsepalum dolficicum, Anacardium occidentale) showed good α-amylase inhibitory effect and can act as good anti-diabetic agents and therefore at least supports their use in the treatment of Diabetes mellitus. The antioxidant activities seen in these plants need to be further explored in synergy with the possible anti-diabetic properties of the plants in the treatment of DM.
TABLE OF CONTENTS
Title page i
Certification ii
Dedication iii
Acknowledgements iv
Table of contents v
List of Tables viii
List of Figures ix
Abstract x
CHAPTER ONE
1.0 INTRODUCTION 1
1.1. Background of this study 1
1.2. Research justification 3
1.3. Research objective 3
CHAPTER TWO
2.0 LITERATURE REVIEW
2.1. Diabetes mellitus 4
2.1.1.
Classification of Diabetes
mellitus 4
2.1.2.
Oral hyperglycemia drugs 6
2.1.3.
Polyphenols 9
2.2.1. Phenolic acids 9
2.2.2. Flavonoids 9
2.3. Free radicals and
oxidative stress 10
2.4. Hyperglycemia and
oxidative stress 10
2.5. Antioxidant
activities of plants 11
2.6. Thin-layer
chromatography 11
2.7. Medicinal plants
investigated in the study 12
2.7.1. Anacardium occidentale 12
2.7.2. Anthocleista djalonensis 12
2.7.3. Ceiba pentandra 13
2.7.4. Chlorophora excelsa 14
2.7.5. Loranthus micranthus 15
2.7.6. Persea americana 16
2.7.7. Picralima nitida 18
2.7.8. Strophanthus hispidus 18
2.7.9. Synsepalum dolficicum 19
CHAPTER THREE
3.0 MATERIALS AND METHODS
3.1. Plant materials 21
3.2. Plant collection and
extract preparation 21
3.3. Quantitative
determination of polyphenols 21
3.3.1. Determination of
phenolic acid content 21
3.3.2. Determination flavonoid
content 22
3.3.3 Determination of plant
metabolites using Thin layer Chromatography (TLC) profiling 24
3.4. In vitroantioxidant activity of plant extracts 23
3.4.1. Ferric
reducingantioxidant power (FRAP) 23
3.4.2. DPPH radical
scavenging activity 24
3.4.3. ABTS radical
scavenging Activity 24
3.5. Alpha-amylase
inhibitory activity 24
3.5.1. Preparation of enzyme solution 24
3.5.2. Preparation of plant extract and enzyme
substrate solution 24
3.5.3. Calibration plot for D-maltose 24
3.5.4. Test for α-Amylase inhibitory activity 25
3.6. Antibacterial
inhibition of plant extracts using disc susceptibility testing 26
CHAPTER FOUR
4.0 RESULTS
4.1.1.
Ethnobotanical information 27
4.2. Polyphenol content of
the plant extracts 28
4.3. Antioxidant
properties of plant extracts 29
4.4. Alpha amylase
inhibitory activities 32
4.5. Thin layer
chromatography (TLC) analysis 33
4.6. Antibacterial studies
of the plant extracts 34
CHAPTER FIVE
5.1.DISCUSSION
5.2.CONCLUSION 38
5.3.RECOMMENDATION 39
5.4.Certificate and abstract note of scientific presentation 40
REFERENCES
LIST OF TABLES
Table 1: Plants used for the treatment of DM in Imo and Abia State,
South East, Nigeria 30
Table 2: FRAP radical scavenging
activity of methanol extract of plant samples 32
Table 3: Percentage inhibition of the α-Amylase inhibition activity
of plant samples tested 32
Table 4: Thin Layer Chromatography retention factor using
Chloroform, Methanol and acetic acid in a ratio of 5:4:1 33
Table 5: Thin Layer Chromatography retention factor using Toluene,
Ethyl acetate and acetic acid in a ratio of 5:4:1 33
Table 6: Table showing the zone of inhibition (mm) of four test
organisms using disk susceptibility testing method 34
LIST OF FIGURES
Figure 1: A diagram showing the pathogenesis of Type 2
diabetes 6
Figure 2: Anacardium
occidentale 13
Figure 3: Anthocleista djalonensis 14
Figure 4: Ceiba pentandra 14
Figure 5: Chlorophora excelsa 15
Figure 6: Loranthus micranthus 17
Figure 7: Persea americana 18
Figure 8: Picralima nitida 19
Figure 9: Strophanthus
hispidus 20
Figure 10: Synsepalum
dolficicum 21
Figure 11: Chart showing the total phenolic content of the plants
studied 32
Figure 12: Chart showing the total flavonoid content of the plants
studied 32
Figure 13: Chart showing the DPPH radical scavenging activities of
the plants studied (IC50 value) 33
Figure 14: Chart showing the ABTS radical scavenging activities of
the plants studied (IC50 value) 34
CHAPTER ONE
INTRODUCTION
1.1 Background of this study
Diabetes mellitus (DM) is a chronic disease caused by inherited
and/or acquired deficiency in production of insulin by the pancreas, or by the
ineffectiveness of the insulin produced. Such a deficiency results in increased
concentrations of glucose in the blood, which in turn damage many of the body's
systems, in particular the blood vessels and nerves. The global prevalence of
the disease is alarmingly increasing partly due to modern lifestyle through
increasing consumption in high carbohydrate diets resulting to postprandial hyperglycemia
(Tappy and Le, 2010). The high increase of blood glucose is partly dependent on
the carbohydrate –hydrolyzing enzymes which are α-amylase and α-glucosidase.
Uncontrolled diabetic hyperglycemia precipitates other factors that accelerate
the progression of diabetic complications mostly micro-vascular complications
associated with neuropathy, nephropathy and retinopathy (Oishi et al., 2006). In Nigeria, the number of
people suffering from this disease has been escalated and now predicted to
reach 10 million of people. It is has been reported that Nigeria is a leading
country where diabetes mellitus is prevalent in Africa (Oyedemi et al., 2010). This is an early
indication for the search of alternative and complementary medicine to the
current oral hypoglycemic drugs which have been identified with adverse
effects.
Presently, there are three forms of diabetes mellitus: Type 1 DM also called
insulin-dependent diabetes mellitus in which the pancreas fails to produce the
insulin which is essential for survival. This form of DM develops most
frequently in children and adolescents, but is being increasingly noted later
in life. Type 2 DM also
known as non-insulin-dependent diabetes mellitus emerges from the inability of
target tissues to respond properly to the insulin action produced by pancreatic
β-cells. This type of DM is much more common and accounts for 90% of all
diabetes cases worldwide. It occurs most frequently in adults, but is being
noted increasingly in adolescents as well. Gestational diabetes mellitus (GDM) resembles type 2 DM in
several respects, involving a combination of relatively inadequate insulin
secretion and responsiveness. It occurs in about 2–10% of all pregnancies and
may improve or disappear after delivery (NDIC, 2011). Other forms of
diabetes mellitus includes Congenital diabetes, which is due to genetic defects
of insulin secretion, cystic
fibrosis-related diabetes, steroid diabetes induced by high doses
of glucocorticoids, and several forms of monogenic diabetes.
The limitation of existing modern medicines due to their side
effects such as diarrhea, flatulence, liver damage and others has necessitated
serious search on alternative therapy with anti-hyperglycemic properties with
less or no side effects especially natural products from plant origin and
polyphenolic rich diets (Ibrahim and Islam, 2014). Plantbaseddrugs are
well-known alternative medicine with cost effective comparable to westernized
medicine. According to world ethnobotanical information reports, more than 400
species of plants have been reported to display hypoglycemic effects, but only
a few of them have been investigated (Miura et
al., 2002)and the World Health Organization has recommended that much
research should be done on hypoglycemic plants (WHO, 2002).
Unfortunately, Nigeria is a country richly blessed with flora but
most of the medicinal plants used traditionally in the treatment of DM lack
scientific data to support their claims. Therefore, this study was conducted to
determine the phenolic content, alpha amylase inhibitory properties and
antioxidative potential of these herbal therapies used in the South Eastern
traditional medicine for the treatment of DM.
1.2. RESEARCH JUSTIFICATION
In Nigeria, the number of people suffering from diabetes mellitus is
believed to be increasing steadily. There are several medicines available in the
market to treat diabetes mellitus but no drug is found to be fully effective
and safe. However, plants and plant products have proven to be effective and
safe in the treatment of diabetes mellitus. This work is being carried out to
determine the various properties of these plants that makes them effective and
safe in the treatment of diabetes mellitus.
1.3. RESEARCH OBJECTIVE
This work is being carried out to determine the various properties
of these plants that makes them effective and safe in the treatment of diabetes
mellitus.
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