Effect Of Ethanol Extract Of Euphorbia Kamerunicus On Potassium Bromate-Induced Toxicity In Albino Rats
ABSTRACT
This study
investigated the modulatory effect of ethanol extract of Euphorbia kamerunicus on potassium bromate-induced toxicity in
Albino rats. The crude ethanol
extract was subjected to phytochemical and, GC-MS analyses and in vitro antioxidant and acute toxicity
evaluations. Results obtained following phytochemical studies of the crude
extract revealed the presence of alkaloids, saponins, tannins, terpenes,
flavonoids, phenols, cardiac glycosides and steroids in various amounts.
Alkaloids were the most abundant (21.63±0.15%) while cardiac glycosides
were the least (4.60±0.26%). GC-MS analysis revealed the presence of 33
compounds in the crude extract with oleic acid as the most abundant (12.54%)
and 2-Propenoic acid the least (0.20%). Other compounds with high abundance in
the crude extract were 11-Octadecenoic acid (8.69%), Butyl 9-tetradecenoate
(8.63%), n-Decanoic acid (6.98%), 1,3-Dioxolane (9.31%), 6-Octadecenoic acid
(7.17%), Methyl stearate (4.27%), 2-Trifluoroacetoxypentadecane (4.54%) and
Hexadecanoic acid (3.74%). The crude extract was also fractionated into five
fractions. Investigation of the effects of the extract in potassium
bromate-induced toxicity was carried out in two phases, first on the crude and then
on the fractions. For the crude extract, 30 rats randomly assigned to 6 groups
of five rats were used. The rats were treated according to the order: group 1
(normal control with no treatment), group 2 (potassium bromate, 100 mg/kg
only), group 3 (200 mg/kg of crude extract + potassium bromate, 100 mg/kg),
group 4 (400 mg/kg of crude extract + potassium bromate, 100 mg/kg), group 5
(800 mg/kg of crude extract + potassium bromate, 100 mg/kg) and group 6 (100
mg/kg vitamin C + potassium bromate, 100 mg/kg). Treatment lasted 28 days
before animals were sacrificed for haematological and biochemical analyses. In
the second phase using the fractions, treatments for groups 1-3 were repeated
but groups 4-8 were treated with extract fractions 1-5 for 28 days before
sacrifice and analyses of collected samples. Results of in vitro antioxidant
activities showed significant nitric oxide and DPPH scavenging activities and
mild ferric reducing antioxidant power activity. Acute toxicity value obtained
for potassium bromate in rats was 346.41 mg/kg body weight while that of the
crude extract was greater than 5000 mg/kg. Results of liver function parameters
showed significantly higher AST, ALT and ALP activities in the group
administered only potassium bromate when compared with those co-treated with
the extract. The crude extract also significantly inhibited anomalies observed
in total protein and serum bilirubin values due to potassium bromate
intoxication. Higher levels of urea, uric acid and creatinine due to potassium
bromate were also significantly lowered in the extract treated groups
(p<0.05). Lipid profile values were not significantly altered following
treatment with potassium bromate and treatment with the extract (p>0.05),
but antioxidant parameters including GSH, GPx, SOD and catalase significantly
depreciated with concurrent rise in the bromate only group but ameliorated in
the groups treated with the extract (p<0.05). The fall in the values of
haematological parameters were also significantly up regulated in the
crude-extract-treated groups. Elevated values of cardiac parameters (lactate
dehydrogenase, creatine phosphokinase and cardiac troponin) due to bromate
intoxication were significantly lowered
in groups treated with the crude extract and fractions (p<0.05).
Inflammatory markers (interleukin 1b, prostaglandin E2 and tumor necrosis
factor) were also lowered significantly (P<0.05). Of all the fractions
evaluated, fraction 4 had the higher activities than the other fractions and
produced effects similar to that of the crude extract. Cactus plant extract may
be of value in the management of potassium bromate-induced systemic toxicity
and could be a potential source of control agent for oxidative stress-induced
diseases caused by environmental oxidants.
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 xiv
Abstract xv
CHAPTER
ONE
INTRODUCTION 1
1.1
Background
of the study 1
1.2
Statement
of the problem 4
1.3
Justification
for the study 6
1.4
Aim of the study 6
1.5
Objectives
of the study 6
CHAPTER TWO
LITERATURE
REVIEW 8
2.1 Potassium
bromate: an overview 8
2.1.1 Physicochemical
properties 9
2.1.2 Uses
and action 10
2.1.2.1 Toxicity
and Safety 11
2.1.3. Potassium bromated in bread making 16
2.2 Systemic Targets For Potassium Bromate
Induced Toxicity 19
2.2.1 Potassium bromate nephrotoxicity 20
2.2.1.1 Pathophysiologic effect
of KBrO3 in the Liver 23
2.2.2 Potassium bromate induced-oxidative stress 24
2.2.3 Potassium bromate-induced hematological
alterations 26
2.2.4 Potassium bromate-induced cardiac
hypertrophy 27
2.2.5 Pathophysiologic effect of KBrO3
in Other Tissues 27
2.3 Mechanism Action of Potassium Bromate
Toxicity 29
2.4 Effects of Plant-Derived Antioxidants 31
2.4.1 Antioxidants 33
2.4.2 Potassium Bromate and Inflammatory Bowel
Disease 33
2.5 Biology
of Euphorbia kamerunicus 35
2.5.1 Medicinal
benefits 36
CHAPTER THREE
MATERIALS
AND METHODS 37
3.1 Materials (Equipment and Reagents) 37
3.2. Collection and Drying of Plant Materials 37
3.2.1 Preparation of Extract 38
3.3. Qualitative
Phytochemical Study of the Extract 38
3.4. Quantitative Phytochemical Tests 40
3.5. In Vitro Antioxidant
Evaluation of the Extract 44
3.6. Determination of Phytochemical Composition of crude cactus
Extract by Gas Chromatography – Mass Spectrometry (Gc-Ms) 46
3.7. Animals 48
3.8 Acute Toxicity Studies 48
3.8.1 Acute Toxicity Evaluation of Potassium Bromate 49
3.8.2 Acute Toxicity Evaluation of Cactus Plant Extract 49
3.9. Experimental Design 50
3.10. Assessment of Haematological Parameters 51
3.11. Assessment of Liver Function Parameters 52
3.12 Assessment of Renal Function Parameters 56
3.13 Lipid Profile Parameters 60
3.14 Assessment of Antioxidant Parameters in
Liver Tissues 63
3.15. Estimation of cardiac parameters (LDH, CPK and cardiac
troponin) 66
3.16. Estimation of inflammatory cytokines (tumor necrotic
factor and
interleukin-1b) 70
3.17 Liver And Kidney Histopathology 74
3.18
Bioassay-Guided Fractionation of Cactus Plant Extract
Using
Chromatographic Techniques 75
3.19
Statistical analysis 80
CHAPTER
FOUR
RESULTS 82
4.1 Results of Phytochemical Evaluation 82
4.1.1 Qualitative phytochemical result 82
4.1.2 Quantitative phytochemical result 84
4.2 Result
of Gc-Ms Analysis of Cactus Plant Crude Extract 86
4.3 Antioxidant Activity Assay 90
4.3.1 DPPH free radical scavenging activity 90
4.3.2 Ferric reducing antioxidant potential (FRAP)
assay 92
4.3.3 Nitric oxide (NO) radical scavenging
activity 94
4.4 Results
of Acute Toxicity Evaluation 96
4.4.1 Result of acute toxicity evaluation of
potassium bromate 96
4.4.2 Acute toxicity report of cactus plant
extract 98
4.5 Liver Function Profile 100
4.6 Serum Renal Profile 103
4.7 Serum Lipid Profile Result 107
4.8 Antioxidant Assay 109
4.9 Haematological Profile 112
4.10 Evaluation of Fractions for Cactus 115
4.10.1 Effects of
fractions on haematological parameters 115
4.10.2 Effects of fractions on liver function
parameters 119
4.10.3 Effects of fractions on renal function
parameters 122
4.10.4 Effects of fractions on lipid profile
parameters 125
4.10.5 Effects of fractions on
antioxidation enzymes 130
4.10.6 Effects of fractions on cardiac biomarkers 134
4.10.7 Effects of fractions on carcinoma biomarkers 136
CHAPTER FIVE
DISCUSSION 140
5.1 Phytochemicals 140
5.2 Antioxidant Potential 142
5.3 Liver Function Profile 145
5.4 Lipid Profile 147
5.5 Renal Function 149
5.6 Hematological
Evaluation 151
5.7 Acute
Toxicity 153
5.8 Antioxidants 154
5.9 Cardiac Markers 156
5.91 Evaluation of Fractions 157
5.92 Inflammatory Markers 158
5.93 Conclusion 160
5.94 Recommendations 160
5.95 Suggestion for Further
Study 161
References 162
LIST OF TABLES
Table 4.1a: Qualitative phytochemical components of ethanol extract
of cactus 83
Table 4.1b: Qualitative phytochemical components of ethanol extract
of cactus 85
Table 4.2: GC-MS crude extract 88
Table 4.3a: Acute toxicity evaluation of
potassium bromate phase 1 97
Table 4.3b: Acute
toxicity evaluation of potassium bromate phase 2 97
Table 4.4a: Acute toxicity evaluation of ethanol extract of cactus phase 1 99
Table 4.4b: Acute toxicity evaluation of ethanol extract of cactus phase 2 99
Table 4.5: Effects of the ethanol extract of cactus on
some serum
biochemical parameters treated exposed
to potassium
bromate 102
Table 4.6: Effects of the ethanol extracts of cactus on plasma urea, creatinine, uric acid and electrolyte levels
in potassium bromate intoxicated Rats. 105
Table 4.7: Effects of the ethanol extract of cactus on some lipid
profile of
potassium bromate intoxicated Rats. 108
Table 4.8: Effects of ethanol extract of cactus
on protective/antioxidant parameters. 111
Table 4.9: The effects of the extract cactus on some
haematological parameters
induced with potassium bromate
intoxicated Rats. 114
Table 4.10: Effect of 5 different fractions and crude extract of cactus
on concentration of some haematological
indices. 117
Table 4.11: Effects of five different
fractions and crude extract of cactus
on concentration of selected
liver function parameters. 120
Table 4.12: Effects of five different fractions and crude extract of
cactus on selected renal function parameters. 125
Table 4.13: Effect of 5 different fractions
and crude extract of cactus
on
selected lipid parameters. 128
Table 4.14: Effects of five different fractions and crude extract of cactus
on redox status of
potassium bromate intoxicated Rats 132
Table 4.15: Effects of five different fractions and crude extract of cactus
on some cardiac
biomarkers of potassium bromate intoxicated Rats.136
Table 4. 16: Effects of five different
fractions and crude extract of cactus
plant
on the concentration levels some carcinoma biomarkers exposed
to
potassium bromate. 139
LIST OF FIGURES
Figure 2.1 Structure
of the Potassium Bromate Molecule 10
Figure 4.1: GC-MS chromatogram showing 33 compounds
in cactus
plant crude extract. 87
Figure 4.2a: DPPH
free radical scavenging antioxidant activity. 91
Figure 4.2b: Ferric
reducing antioxidant power (FRAP) assay. 93
Figure 4.2c: Nitric
oxide radical scavenging activity. 95
0.0
ATASIE, A (2023). Effect Of Ethanol Extract Of Euphorbia Kamerunicus On Potassium Bromate-Induced Toxicity In Albino Rats. Repository.mouau.edu.ng: Retrieved Nov 23, 2024, from https://repository.mouau.edu.ng/work/view/effect-of-ethanol-extract-of-euphorbia-kamerunicus-on-potassium-bromate-induced-toxicity-in-albino-rats-7-2
ATASIE, ATASIE. "Effect Of Ethanol Extract Of Euphorbia Kamerunicus On Potassium Bromate-Induced Toxicity In Albino Rats" Repository.mouau.edu.ng. Repository.mouau.edu.ng, 19 Jul. 2023, https://repository.mouau.edu.ng/work/view/effect-of-ethanol-extract-of-euphorbia-kamerunicus-on-potassium-bromate-induced-toxicity-in-albino-rats-7-2. Accessed 23 Nov. 2024.
ATASIE, ATASIE. "Effect Of Ethanol Extract Of Euphorbia Kamerunicus On Potassium Bromate-Induced Toxicity In Albino Rats". Repository.mouau.edu.ng, Repository.mouau.edu.ng, 19 Jul. 2023. Web. 23 Nov. 2024. < https://repository.mouau.edu.ng/work/view/effect-of-ethanol-extract-of-euphorbia-kamerunicus-on-potassium-bromate-induced-toxicity-in-albino-rats-7-2 >.
ATASIE, ATASIE. "Effect Of Ethanol Extract Of Euphorbia Kamerunicus On Potassium Bromate-Induced Toxicity In Albino Rats" Repository.mouau.edu.ng (2023). Accessed 23 Nov. 2024. https://repository.mouau.edu.ng/work/view/effect-of-ethanol-extract-of-euphorbia-kamerunicus-on-potassium-bromate-induced-toxicity-in-albino-rats-7-2