SCREENING OF SENNA ALATA LEAVES FOR BIOCHEMICAL AND HYPOGLYCEMIC ACTIVITY
ABSTRACT
The comparative effects of Sennaalata leaves extract and sulfonylurea drug Glibenclamide on biochemical indices were evaluated in Wistar rats. The design consisted of 4 groups of 6 rats each. Group 1 (NC) and 2 (DC) received placebo, groups 3 (SA) received ethanolic extract of Sennaalata leaves equivalent of 250mg/kg body weight, while group 4 (GB) was gavaged with 5mg/kg body weight of Glibenclamide each administered 12 hourly-twice daily. Treatment lasted for 4weeks (28 days), during which fasting blood glucose of the rats were monitored on the 1st, 7th, 13th, 19th, 25th and 28th day respectively, after which the animals were sacrificed and blood collected for biochemical and haematological analysis. Results of biochemical indices indicated significant (p<0.05) hypoglycemic activity in both treated groups compared to the diabetic group. The reduction in fasting blood glucose for SA (80.6%) and GB (84%) over the treatment period was significant (P<0.05) compared to DC (45%). Hepatotoxicity indices (ALT, AST) showed significant (P<0.05) reduction in AST levels of GB (18.14±5.45) compared to DC (165.62±14.95), although SA (271.17±26.31) was markedly increased when compared to DC. ALT activity was significantly (P<0.05) increased in SA and GB compared to DC. Treatments produced a profound decrease in total proteins in treated animals compared with the controls but the decrease was significant (P<0.05) only when compared with DC. Body weight result showed significant (P<0.05) weight gain in both SA and GB diabetic treated groups compared to the DC diabetic untreated group. In conclusion this study clearly indicates that Sennaalata extract may reverse hyperglycemic states and ameliorate the associated complications in diabetes mellitus and compares favourably with standard drug.
TABLE OF CONTENTS
COVER PAGE i
CERTIFICATION ii
DECLARATION iii
ACKNOWLEDGEMENT iv
ABSTRACT vi
TABLE OF CONTENTS viii
LIST OF TABLES xiii
LIST OF FIGURE xiv
LIST OF ABBREVIATIONS xv
CHAPTER ONE: INTRODUCTION
1.1 Background information 1
1.2 Statement of problem 3
1.3 Justification of the study 4
1.4 Aims and objectives of the study 6
1.5 Scope of the study 6
CHAPTER TWO: LITERATURE REVIEW
2.1 SennaAlata 8
2.1.1 Botanical Classification 9
2.1.2 Description 9
2.1.3 Utilization and Importance 10
2.1.3.1 Medicinal Importance 11
2.1.4 Toxicity 12
2.2 Diabetes Mellitus 13
2.2.1 Definition 13
2.2.2 Classification 14
2.2.2.1 Type 1 Diabetes Mellitus 16
2.2.2.2 Type 2 Diabetes Mellitus 17
2.2.2.3 Maturity Onset Diabetes of the Young 18
2.2.2.4 Gestational Diabetes Mellitus 20
2.2.3 Hyperglycemia 21
2.2.3.1 Non-Enzymatic Glycation 22
2.2.3.2 Other Mechanism 23
2.2.4 Glucose Toxicity 24
2.2.5 Diabetes and Macro vascular Diseases 25
2.2.6 Micro vascular Complications of Diabetes 27
2.2.6.1 Diabetic Retinopathy 27
2.2.6.2 Diabetic Nephropathy 29
2.2.6.3 Diabetic Neuropathy 30
2.3 Chemotherapy of Diabetes 32
2.3.1 Insulin 32
2.3.1.1 Protein Structure of Insulin 35
2.3.1.2 Synthesis of Insulin 36
2.3.1.3 Mechanism of Action 38
2.3.2 Oral Hypoglycemic Agents 40
2.3.2.1 Sulfonylurea 40
2.3.2.2 Biguanides 43
2.3.2.3 Thiazolidinediones 44
2.3.2.4 α-Glucosidase Inhibitors 46
2.4 Streptozotocin (STZ) Diabetes Mechanism 46
CHAPTER THREE: MATERIALS AND METHODS
3.1 Materials 48
3.1.1 Chemicals and reagents 48
3.1.2 Equipment and glass wares 48
3.2 Methods 49
3.2.1 Plant materials 49
3.2.1.1 Collection of plant materials 49
3.2.2 Animals 50
3.2.2.1 Experimental design 50
3.2.2.2 Induction of experimental diabetes 51
3.3 Extract and drug administration 51
3.4 Experimental procedure 51
3.5 Collection of samples for analysis 52
3.6 Data and Statistical Analysis 53
3.7 Estimation of Biochemical Parameters 55
3.7.1 Total Protein Determination 55
3.7.2 Albumin 56
3.7.3. Total Bilirubin Determination 57
3.7.4 Urea Determination 58
3.7.5 Creatinine Determination 60
3.8 Estimation of Lipid Profile 61
3.8.1 HDL Cholesterol 61
3.8.2 Cholesterol 63
3.8.3 Triglycerides 65
3.8.4 Estimation of LDL- Cholesterol Concentration 67
3.8.5 Estimation of VLDL – cholesterol concentration 67
3.9 Estimation of Liver Enzyme Activity 67
3.9.1 Alanine Aminotransferase (ALT) 67
3.9.2 Aspartate Aminotransferase (AST) 63
3.9.3 Alkaline Phosphatase (ALP) 63
3.10 Estimation of Electrolyte Balance 63
3.10.1 Sodium 64
3.10.2 Estimation of Serum Chloride Concentration 64
3.10.3 Phosphorus 64
3.10.4 Potassium 64
3.15 Phytochemical Screening 65
CHAPTER FOUR: RESULT
4.1 Anti-Diabetic Activity of Extracts of Sennaalata
leaves on Rats 66
4.2 Effect of Treatment on Body weight 66
4.3 Effect of Treatment on lipid profile 66
4.4 Effect of Treatment on serum enzyme activity 67
4.5 Effect of Treatment on Serum Protein 67
4.6 Effect of Treatment on Serum Electrolyte 91
4.7 Phytochemical Screening 91
CHAPTER FIVE: DISCUTION, CONCLUSION ANDRECOMMENDATION
5.1 Discussion 97
5.1.1 Total Body Weight 97
5.1.2 Blood Glucose 98
5.1.3 Total Protein, Albumin and Globulin 99
5.1.4 Serum Lipid Profile 99
5.1.5 Serum Enzymes 100
5.2 Summary and Conclusion 101
5.3 Contribution to Knowledge 102
REFERENCES 103
APPENDIX 123
LIST OF TABLES
Table 1Animal grouping and treatment schedule 56
Table 2Effect of crude extract of Sennaalata leaves on
STZ-induced Diabetic rat 69
Table 3 Effect of crude extract of Sennaalata leaves on Body
Weight of STZ-induced Diabetic rat treated for 28 days 70
Table 4 Effect of crude extract of Sennaalata leaves on Lipid
Profile of STZ-induced Diabetic rat 72
Table 5 Effect of crude extract of Sennaalata leaves on Serum
enzyme activities of STZ-induced Diabetic rat 75
Table 6 Effect of crude extract of Sennaalata leaves on Serum
Protein of STZ-induced Diabetic rat 77
Table 7 Effect of crude extract of Sennaalata leaves on Serum Electrolytes of STZ-induced Diabetic rat
Table 8Phytochemical Screening of Sennaalata
LIST OF FIGURE
Figure 1Sennaalata plant
Figure 2 Fasting Blood Glucose levels in different experimental groups 11
Figure 3 Effect of Sennaalata leaves extract on Body weight
of Rats 41
Figure 4 Effect of Extracts on lipid profile 34
Figure 5 Effect of Extract on Serum Protein
ABBREVIATIONS
A/G - Albumin Globulin ratio
ALP - Alkaline phosphatase
ALT - Alanine transaminase
AST - Aspartate transaminase
CDC - Centres for Disease Control
DMSO - Dimethylsulphoxide
HDL - High Density Lipoprotein
LDL - Low density lipoprotein
TC - Total Cholesterol
TG - Triglyceride
VLDL - Very Low Density Lipoprotein
WHO - World Health Organization
.