Assessment Of The Cardioprotective Potential Of The Ethanol Leaf Extract Of Pterocarpus santalinoides In Wistar Rats
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ABSTRACT
Cardiotoxicity has become a major challenge for cancer patients undergoing chemotherapy. Examples of potentially cardiotoxic anticancer agents are 5-fluorouracil and anthracyclines. The leaves of Pterocarpus santalinoides have been shown to possess among other pharmacological properties, the ability to lower serum levels of low density lipoprotein cholesterol (LDL-C) which suggests a cardio protective potential. This study was aimed at experimenting the cardioprotective potential of the ethanol leaf extract against cardiotoxicity side effect of 5-fluorouracil in wistar rats. The study was done in two phases. 36 albino rats were employed for the first phase of the study, they were separated into 6 groups (groups 1, 2, 3, 4, 5 and 6) of 6 animals which received (distilled water, distilled water, 25 mg/kg Aspirin, 34.54 mg/kg crude extract, 69.28 mg/kg crude extract and 103.92 mg/kg crude extract for groups 1, 2, 3, 4, 5 and 6 respectively) orally for a period of 14 days. On the 15th day, groups 2, 3, 4, 5, and 6 were given 5-fluorouracil (150 mg/kg) via intra-peritoneal route to induce cardiotoxicity. After a period of 24 hours, the animals were sacrificed, the sera were analyzed for troponin T, CK-MB, Lipid profile, the whole blood samples were subjected to haematology while the hearts were subjected to histopathological scrutiny, the result obtained revealed significant () increase in HDL-C and decrease in LDL-C, Troponin T and CK-MB across groups compared to positive control which indicates cardioprotective potential, hence the second phase was conducted. In the second phase, the crude extract was fractionated into n-butanol, ethyl acetate and chloroform fractions, 36 albino wistar rats were also used for the study. They were also separated into 6 groups of 6 animals each. The animals were equally given treatments (distilled water, distilled water, 25 mg/kg Aspirin, 69.28 mg/kg n-butanol fraction, 69.28 mg/kg ethyl acetate fraction and 69.28 mg/kg chloroform fraction for groups 1, 2, 3, 4, 5 and 6 respectively) orally for 14 days. On the 15th day, groups 2, 3, 4, 5 and 6 were given 5-FU (150 mg/kg) via intraperitoneal route to induce cardiotoxicity. The animals were sacrificed after 24 hours, and the sera analyzed for troponin T level, CK-MB activity; lipid profile; urea, creatinine and electrolytes concentrations; ALP, AST and ALT activity; malondialdehyde concentration, catalase, glutathione peroxidase, superoxide dismutase activity; the whole blood samples were haematologically analyzed and the heart, kidney and liver tissues were histologically studied. Result revealed significant decrease () in TnT, CK-MB, LDL-C, TG, WBC, platelets, ALP, AST, ALT, urea, creatinine, Na+, K+, Cl-, HCO-3 and significant (p<0.05) increase in HDL-C, RBC, Ca2+, catalase, SOD and GSPx activity across groups as compared to the positive control. Histopathological results revealed that the low and medium dose of crude extract and chloroform fraction yielded more effective protection from cardiotoxicity more than the other doses and fractions. The crude ethanol extract of P. santalinoides and its fractions conferred on the albino rats appreciable protection against 5-FU induced cardiotoxicity with the medium dose and chloroform fraction producing the overall highest cardioprotective effects.
TABLE OF CONTENTS
Title Page i
Declaration ii
Certification iii
Dedication iv
Acknowledgements v
Table of Contents vi
List of Tables xii
List of Figures xiii
List of Plates xii
List of Appendixes xv
List of Abbreviations xvii
Abstract xix
CHAPTER 1: INTRODUCTION
1.1 Background of the Study 1
1.2 Pterocarpus santalinoides (PS) 7
1.3 Scope of the Study 10
1.4 Statement of Problem 11
1.5 Justification of the Study 12
1.6 Relevance of the Study 13
1.7 Aim of the Study 14
1.8 Objectives of the Study 14
CHAPTER 2: REVIEW OF RELATED LITERATURE
2.1 Phytochemical and Proximate Composition of Pterocarpus santalinoides 15
2.1.1 Phytochemistry of Pterocarpus santalinoides 15
2.1.2 Proximate composition of Pterocarpus santalinoides 16
2.2 Pharmacological Activities of Pterocarpus santalinoides 17
2.2.1 Analgesic activity 17
2.2.2 Insecticidal activity 17
2.2.3 Antitrypanosomal activity 19
2.2.4 Haematological effects 19
2.2.5 Hypolipidemic activity 19
2.2,6 Anti-cancer activity 20
2.2.7 Antimicrobial activity 21
2.2.8 Anti-diarrhoeal and antispasmodic activity 22
2.3 Cardioprotective Effect of Aspirin 24
2.4 Anti-Cancer agents and their Cardiotoxicity Potential 25
2.4.1 5-Fluorouracil and other antimetabolites 25
2.4.2 Trastuzumab (Herceptin) 27
2.4.3 Anthracyclines 28
2.4.4 Taxanes 30
2.4.5 Interleukin-2 (IL-2) therapy 31
2.4.6 Tyrosine kinase inhibitors (TKIs) 31
2.4.7 Checkpoint inhibitors immune therapy 32
2.4.8 Angiogenesis inhibitors 33
2.4.9 Radiation therapy 34
2.5 Biomarkers of Cardiotoxicity 35
2.5.1 Cardiac function status 35
2.5.1.1 Troponin 35
2.5.1.2 Creatine kinase (EC 2.7.3.2) 36
2.5.1.3 Lactate dehydrogenase (EC 1.1.1.27) 37
2.5.1.4 Natriuretic peptide 38
2.5.2 Peripheral blood mononuclear cell gene expression profile 40
2.5.3 Lipid profile (cardiovascular risk factor assessment) 41
2.5.3.1 Cholesterol 41
2.5.3.2 Triglycerides 43
2.5.4 Liver function status 43
2.5.4.1 Alkaline Phosphatase (ALP) (EC 3.1.3.1) 44
2.5.4.2 Aminotransferases: ALT (EC 2.6.1.2) and AST (EC 2.6.1.1) 45
2.5.5 Oxidative stress status 47
2.5.5.1 Malondialdehyde (MDA) 47
2.5.5.2 Catalase (EC 1.11.1.6) 48
2.5.5.3 Superoxide dismutase (EC 1.15.1.1) 50
2.5.5.4 Glutathione peroxidase (EC.1.11.1.9) 51
2.5.6 Haematological indices 52
2.5.6.1 Red blood cell (erythrocytes) count and haemoglobin (Hb) 54
2.5.6.2 White blood cell (WBC) count 56
2.5.5.3 Platelet count 56
2.5.7 Kidney function status 57
2.5.7.1 Serum potassium 57
2.5.7.2 Serum sodium 59
2.5.7.3 Serum bicarbonate 60
2.5.7.3 Serum creatinine 61
2.5.7.4 Serum urea 62
CHAPTER 3: MATERIALS AND METHODS
3.1 Materials 64
3.1.1 Chemicals and reagents 64
3.1.2 Equipment 64
3.1.3 Research hypothesis 65
3.2 Methods 65
3.2.1 Plant leaf collection, identification and extraction of
the crude ethanol extract 65
3.2.2 Extract fractionation by partitioning 66
3.2.3 Determination of median lethal dose (LD50) for the crude ethanol
extract 66
3.2.4 Phytochemical screening 68
3.2.4.1 Test for alkaloids 68
3.2.4.2 Test for saponins 68
3.2.4.3 Test for tannins 68
3.2.4.4 Test for flavonoids 69
3.2.4.5 Test for cardiac glycosides 69
3.2.4.6 Test for anthraquinones 69
3.2.5 Experimental design 70
3.2.6 Experimental animals treatment 71
3.2.7 Animal sacrifice and preparation of sera for phase one studies 72
3.2.8 Animal sacrifice and preparation of sera for phase two studies 73
3.3 Biochemical Analysis 75
3.3.1 Estimation of serum troponin-T concentration 75
3.3.2 Estimation of CK-MB concentration 76
3.3.3 Estimation of alkaline phosphatase activity 78
3.3.4 Estimation of serum superoxide dismutase activity 79
3.3.5 Estimation of serum glutathione peroxidase activity 80
3.3.6 Estimation of serum catalase activity 82
3.3.7 Estimation of serum malondialdehyde (MDA) level 83
3.3.8 Estimation of total serum cholesterol 84
3.3.9 Estimation of serum high density lipoprotein cholesterol 85
3.3.10 Estimation of serum level of total triglyceride 86
3.3.11 Estimation of haematological indices 88
3.3.12 Estimation of serum electrolytes level 89
3.3.13 Determination of aspartate amino transferase (AST) activity in serum 90
3.3.14 Estimation of alanine aminotransferase (ALT/SGPT) activity 91
3.3.15 Estimation of urea (BUN) 92
3.3.16 Estimation of serum creatinine concentration 92
3.3.17 Histological procedures 94
3.3.18 Statistical analysis 94
CHAPTER 4: RESULTS AND DISCUSSION
4.1 Results 96
4.1.1 Phytochemical screening 96
4.1.2 Phase one results 98
4.1.2.1 Effect of CELEPS on serum troponin-T concentration and
CK-MB activity in albino rats 98
4.1.2.2 Effect of CELEPS on serum lipid profile in albino rats 99
4.1.2.3 Effect of CELEPS on haematological parameters in albino
rats 102
4.1.2.4 Effect of CELEPS on the histopathology of the heart tissues 104
4.1.3 Phase two results 112
4.1.3.1 The effect of FELEPS on cardiovascular indices in albino rats 112
4.1.3.1.1 The effect of FELEPS on serum troponin T concentration in
albino rats 112
4.1.3.1.2 The effect of FELEPS on serum CK-MB activity in albino
rats 114
4.1.3.2 The effect of FELEPS on serum lipid profile in albino rats 116
4.1.3.3 The effect of FELEPS on liver function status in albino rats 120
4.1.3.3.1 The effect of FELEPS on serum ALP, AST and ALT activity 120
4.1.3.4 The effect of FELEPS on the oxidative stress status in albino
rats 122
4.1.3.4.1 The effect of FELEPS on the serum MDA concentration in
albino rats 122
4.1.3.4.2 The effect of FELEPS on serum levels of oxidative enzymes;
Catalase, Superoxide Dismutase and Glutathione Peroxidase in
Albino Rats 124
4.1.3.5 The effect of FELEPS on the haematological Indices in albino
Rats 125
4.1.3.6 The effect of FELEPS on kidney function status in albino rats 132
4.1.3.6.1 The effect of FELEPS on the serum electrolytes concentration in albino rats 132
4.1.3.6.2 The effect of FELEPS on the serum urea and creatinine
concentration in albino rats 135
4.1.3.7 Histopathological results 137
4.2 Discussion 155
CHAPTER 5: SUMMARY, CONCLUSION AND RECOMMENDATIONS
5.1 Summary 180
5.2 Conclusion 182
5.3 Recommendation 183
References 184
Appendix 220
LIST OF TABLES
3.1 Dose Regimen for Phase One Studies (Treatment with Crude Ethanol
Extract) 70
3.2 Dose Regimen for Phase Two (Treatment with Fractionated Portions of the
Crude ethanol extract) 72
3.3 Sample and Blank Regimen for Determination of SOD activity 79
3.4 Reagent Blank, Standard, Controls and Samples Regimen for Estimation
of Serum Creatinine Concentration 92
4.1 Result of Phytochemical Screening 95
4.2 Effect of CELEPS on Serum of Troponin Concentration and
CK-MB Activity in Albino rats 97
4.3 Effect of CELEPS on Serum Lipid Profile in Albino Rats 99
4.4 Effect of CELEPS on Haematological Parameters in Albino Rats 101
4.5 The Effect of FELEPS on Serum Lipid Profile in Albino Rats 116
4.6 The Effect of FELEPS on Lipid Profile Assessment (cardiovascular risk index) 117
4.7 The Effect of FELEPS on Serum ALP, ALT and AST Activities in Albino Rats 119
4.8 The Effect of FELEPS on the Serum MDA Concentration in Albino Rats 121
4.9 The Effect of FELEPS on Serum Activities of Oxidative Enzymes; Catalase, Superoxide Dismutase and Glutathione Peroxidase in Albino Rats 123
4.10A The Effect of FELEPS on the Haematological Indices in Albino Rats 127
4.10B The Effect of FELEPS on the haematological Indices in albino rats continued 128
4.11 The Effect of Fractions of Ethanol Leaf Extract of P. santalinoides on the Serum Electrolytes in Albino Rats. 131
4.12: The Effect of FELEPS on the Serum Urea and Creatinine Concentration in Albino Rats in Wistar rats 133
LIST OF FIGURES
4.1 The effect of fractions of ethanol leaves extract of p. santalinoides
(FELEPS) on serum troponin levels (ng/ml) in albino rats. 111
4.2 The effect of fractions of ethanol leaf extract of P. santalinoides
(FELEPS) on serum CK-MB activity (u/l) in albino rats 113
LIST OF PLATES
1.1 Pterocarpus santalinoides 7
4.1 Photomicrograph of Heart Tissue of Normal Control Rats (Treated with Distilled Water Only) 104
4.2 Photomicrograph of Heart Tissue Treated with Distilled Water and 150 mg/kg bw 5-Fluorouracil (Positive Control) 105
4.3 Photomicrograph of Heart Tissue Treated with 25 mg/kg bw Aspirin (AS) and 150 mg/kg bw 5-Fluorouracil (Standard Control) 106
4.4 Photomicrograph of Heart Tissue Treated with Low Dose (LD) (34.64 mg/kg bw) of CELEPS and 150 mg/kg bw 5-Fluorouracil (Low Dose) 107
4.5: Photomicrograph of Heart Tissue Treated with Medium Dose (69.28 mg/kg bw) of CELEPS and 150 mg/kg bw 5-Fluorouracil 108
4.6 Photomicrograph of Heart Tissue Treated with High Dose (HD) (103.92 mg/kg bw) CELEPS and 150 mg/kg bw 5-Fluorouracil. 109
4.7 Photomicrograph of the Longitudinal Section of the Distilled Water
(Control) Heart Tissue 134
4.8 Photomicrograph of the Longitudinal Section of the Heart Tissue of the Distilled Water + 150 mg/kg bw 5-Fluorouracil Treated Rats (Positive Control) 135
4.9 Photomicrograph of the Longitudinal Section of the Heart tissue of the 25mg/kg bw aspirin + 150mg/kg bw 5-Fluorouracil Treated Rats (Standard Control) 136
4.10 Photomicrograph of the Longitudinal Section of the Heart Tissue of the N-Butanol Fraction + 150mg/kg bw 5-Fluorouracil Treated Rats (Group 4) 137
4.11 Photomicrograph of the Longitudinal Section of the Heart Tissue of the Ethyl-acetate Fraction + 150mg/kg bw 5-Fluorouracil Treated Rats (Group 5) 138
4.12 Photomicrograph of the Longitudinal Section of the Heart Tissue of the Chloroform Fraction + 150mg/kg bw 5-Fluorouracil Treated Rats (Group 6) 139
4.13 Photomicrograph of the Longitudinal Section of the Kidney Tissue of the Distilled Water Treated Rats (Normal Control) 140
4.14 Photomicrograph of the Longitudinal Section of the Kidney Tissue of the Distilled Water + 150 mg/kg bw Treated Rats (Positive Control). 141
4.15 Photomicrograph of the Longitudinal Section of the Kidney Tissue of the 25mg/kg bw Aspirin + 150mg/kg 5-Fluorouracil Treated Rats (Standard Control) 142
4.16 Photomicrograph of the Longitudinal Section of the Kidney Tissue of the n-Butanol Fraction + 150 mg/kg bw Treated Rats (Group 4) 143
4.17 Photomicrograph of the Longitudinal Section of the Kidney Tissue of the Ethyl-acetate Fraction + 150 mg/kg bw Treated Rats (Group 5) 144
4.18 Photomicrograph of the Longitudinal Section of the Kidney Tissue of the Chloroform Fraction + 150 mg/kg bw Treated Rats (group 6) 145
4.19 Photomicrograph of the Transverse Section of the Liver Tissue of the Distilled Water Treated Rats (Normal Control). 146
4.20 Photomicrograph of the Transverse Section of the Liver Tissue of the Distilled Water + 150 mg/kg bw 5-Fluorouracil Treated Rats (Positive Control) 147
4.21 Photomicrograph of the Transverse Section of the Liver Tissue of the 25mg/kg bw Aspirin + 150 mg/kg bw 5-Fluorouracil Treated Rats (Standard Control) 148
4.22 Photomicrograph of the Transverse Section of the Liver Tissue of the N-Butanol Fraction + 150 mg/kg bw 5-Fluorouracil Treated Rats (Group 4) 149
4.23 Photomicrograph of the Transverse Section of the Liver Tissue of the Ethyl-acetate Fraction + 150 mg/kg bw 5-Fluorouracil Treated Rats (Group 5) 150
4.24 Photomicrograph of the Transverse Section of the Liver Tissue of the Chloroform Fraction + 150 mg/kg bw 5-Fluorouracil Treated Rats (Group 6) 151
LIST OF APPENDIXES
I Estimation of Serum Troponin 217
II Estimation of Serum CK-MB Concentration 218
III Estimation of Serum Level of Triglyceride (TG) 220
IV Estimation of Total Serum Cholesterol 221
V Estimation of Serum Level of HDL-Cholesterol 221
VI Estimation of Haematological Indices 222
VII Estimation of Serum Malondialdehyde (MDA) Level 223
VIII Estimation of Serum SOD Activity 224
IX Estimation of Serum Glutathione Peroxidase (GSPx) Activity 224
X Estimation of Serum Catalase (CAT) Activity 225
XI Estimation of Alkaline Phosphatase (ALP) Activity 225
XII Estimation of Aspartate Amino Transferase (AST) Activity 226
XIII Estimation of Alanine Amino Transferase (ALT) Activity 226
XIV Estimation of Serum Urea Nitrogen (BUN) Concentration 227
XV Estimation of Serum Creatinine Concentration 227
XVI Preparation of Extracts Doses (mg/kg bw) 228
XVII Preparation of Aspirin 25 Mg (mg/kg bw) 228
XVIII Calculation of 5-Fluorouracil Doses (mg/kg bw) 228
LIST OF ABBREVIATIONS
5-FU 5-Fluorouracil
ACE Angiotensin Converting Enzyme
ALP Alkaline Phosphatase
ALT Alanine Transaminase
AST Aspartate Transaminase
BUN Blood Urea Nitrogen
CAD Coronary Artery Disease
CAT Catalase
CBC Complete Blood Count
CDC Center for Disease Control and Prevention
CELEPS Crude ethanol extract of Pterocarpus santalinoides
CHF Congestive Heart Failure
CKD Chronic Kidney Disease
CK-MB Creatine Kinase-MB Fraction
CVD Cardiovascular Disease
DNPH Dinitrophenyl Hydrazone
FBC Full Blood Count
FDA Food and Drug Administration
FELEPS Fractions of Ethanol Extract of Pterocarpus santalinoides
GGT Gamma-Glutamyl Transferase
GSPx/GPx Glutathione Peroxidase
H &E Hematoxylin and Eosin
Hb/HGBS Haemoglobin
HDL-C High Density Lipoprotein Cholesterol
HER2 Human Epidermal Growth Factor Receptor 2
HF Heart Failure
HIV Human Immunodeficiency Virus
LD50 Lethal Dosage
LDL-C Low Density Lipoprotein Cholesterol
LV Left Ventricle
LVD Left Ventricular Dysfunction
LVEF Left Ventricular Ejection Fraction
Mabs Monoclonal Antibodies
MDA Malondialdehyde
MEPS Methanol Leaf Extract of Pterocarpus santalinoides
MI Myocardial Infarction
NO Nitric Oxide
NSAID Nonsteroidal Anti-inflammatory Drug
PMBC Peripheral blood Mononuclear Cells
PS Pterocarpus santalinoides
RBC Red Blood Cell
SGOT Serum Glutamic Oxaloacetic Transaminase
SGPT Serum Glutamic Pyruvic Transaminase
SOD Superoxide Dismutase
TC Total Cholesterol
TKIs Tyrosine Kinase Inhibitors
TnI Troponin I
TnT Troponin T
UA Uric Acid
VEGF Vascular Endothelial Growth Factor
VLDL Very Low Density Lipoprotein
WBC White Blood Cell
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APA
ONWUCHEKWA, & UCHECHI, B. (2023). Assessment Of The Cardioprotective Potential Of The Ethanol Leaf Extract Of Pterocarpus santalinoides In Wistar Rats. Michael Okpara University of Agriculture. Retrieved June 7, 2026, from http://repository.mouau.edu.ng/works/assessment-of-the-cardioprotective-potential-of-the-ethanol-leaf-extract-of-pterocarpus-santalinoides-in-wistar-rats-7-2
MLA
ONWUCHEKWA, and BLESSING UCHECHI. "Assessment Of The Cardioprotective Potential Of The Ethanol Leaf Extract Of Pterocarpus santalinoides In Wistar Rats." Michael Okpara University of Agriculture, 14 Aug. 2023, http://repository.mouau.edu.ng/works/assessment-of-the-cardioprotective-potential-of-the-ethanol-leaf-extract-of-pterocarpus-santalinoides-in-wistar-rats-7-2. Accessed June 7, 2026.
Chicago
ONWUCHEKWA, and BLESSING UCHECHI. "Assessment Of The Cardioprotective Potential Of The Ethanol Leaf Extract Of Pterocarpus santalinoides In Wistar Rats." Michael Okpara University of Agriculture (2023). Accessed June 7, 2026. http://repository.mouau.edu.ng/works/assessment-of-the-cardioprotective-potential-of-the-ethanol-leaf-extract-of-pterocarpus-santalinoides-in-wistar-rats-7-2