THE EFFECT OF QUININE-ARTESUNATE CO-ADMINISTRATION ON SOME BIOCHEMICAL PARAMETERS IN PLASMODIUM BERGHEI PARASITIZED SWISS ALBINO MICE AND RATS


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TABLE OF CONTENTS

Title Page
ABSTRACT
TABLE OF CONTENTS
ABBREVIATIONS, DEFINITIONS, GLOSSARY AND SYMBOLS

CHAPTER ONE
1.0       INTRODUCTION
1.1       Preamble
1.2       Statement of Research Problem
1.3       Justification
1.4       Aim
1.5       Hypothesis

CHAPTER TWO
2.0       LITERATURE REVIEW
2.1       Malaria
2.2       Prevalence of Malaria
2.3       Transmission
2.4       Signs and Symptoms of Malaria
2.5       Classification of Malaria
2.6       Diagnosis of Malaria
2.7       Life cycle of Malaria Parasite
2.7.1    The Asexual Erythrocytic Phase in Humans
2.7.2    The Sexual Stage in Mosquito
2.8       Malaria and Glucose Metabolism
2.9       Oxidative Stress in Malaria
2.10     Treatment of Malaria
2.11     Quinine
2.11.1  Chemistry
2.11.2  Pharmacokinetics
2.11.3  The Mechanism of Action of Quinine
2.11.4  Resistance
2.11.5  Therapeutic uses
2.11.6  Adverse Effects
2.11.7  Contraindications and Drug Interactions
2.12     Artesunate: Artemisinin Derivative
2.12.1  Mechanism of Action of Artesunate
2.12.2  Combination Therapy
2.13     Ibuprofen
2.14     Drug-Drug Interaction
2.14.1  Quinine-Artesunate Combination

CHAPTER THREE
3.0       MATERIALS AND METHODS
3.1       Laboratory Animals
3.2       Chemicals, Drugs, Reagents and other Materials
3.3       Preparation of Drugs
3.4       Study Design
3.5       Malaria Parasitized Animals
3.6       Glucose Level on Healthy Animals (Uninfected mice)
3.6.1    Oral Glucose Tolerance Test
3.7       Biochemical Parameters Analysis
3.7.1    Liver Function Tests
3.7.2    Oxidative Stress Biomarkers
3.8       Haematological Parameters Analysis
3.9       Organ Body Weight Ratio
3.10     Analgesic Study: Acetic Acid Induced Abdominal Constrictions
3.11     Anti-Pyretic Study: Brewer’s Yeast Induced Pyrexia Model
3.12     Statistical Analysis

CHAPTER FOUR
4.0       RESULTS
4.1       The Effect of Quinine, Artesunate, Quinine-Artesunate Co-administration and their Separate Therapy on Blood Glucose Level in Swiss Albino Mice
4.2       The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Oral Glucose Tolerance Test in Swiss Albino Mice
4.3       The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Liver Function Test of Albino Mice
4.4       The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Oxidative Stress Biomarkers on Swiss Albino Mice
4.5       The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Hematological Parameters in Swiss Albino Mice
4.6       The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Relative Organ Body Weight in Swiss Albino Mice
4.7       The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Random Blood Glucose Level in P. berghei Infected Mice
4.8       The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Fasting Blood Glucose Level in P. berghei infected mice
4.9       The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Liver Function Test in Plasmodium berghei Infected Mice
4.10     The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Oxidative Stress Biomarkers in P. berghei Infected Mice
4.11     The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Relative Organ Body Weight of P. berghei Infected Mice
4.12     The Effect of Quinine-Artesunate Co-administration on Acetic Acid Induced Writhes in Mice
4.13     The Effect of Quinine-Artesunate Co-administration and their Separate Therapy on Brewer’s Yeast Induced Pyrexia in Rats

CHAPTER FIVE
5.0       DISCUSSIONS

CHAPTER SIX
6.0       SUMMARY, CONCLUSION AND RECOMMENDATIONS
6.1       Summary
6.2       Conclusion
6.3       Recommendations
REFERENCES
APPENDIX



ABSTRACT

Malaria has been the leading cause of morbidity and mortality in most Sub-saharan Africa and Southeastern Asia. Combination therapy has been recommended for treatment of malaria and this could lead to drug-drug interaction. This work titled “The effect of Quinine-Artesunate co-administration on some biochemical parameters on healthy and

Plasmodium berghei parasitized Swiss albino mice and rats” was carried out to investigate the effect of Quinine (60 mg/kg)-Artesunate (30 mg/kg) co-administration following 7 days pre treatment. Mice were infected with Plasmodium berghei NK65 strain. Parasitaemia was checked after Geimsa stain and viewed for infected blood cells. Five days of establishment of the infection, they were treated for 7 days. The effect on blood glucose level was evaluated using the glucose oxidase principle; the Liver functions and Oxidative stress were assessed using colorimetric kits. The fasting blood glucose and oral glucose tolerance test of the healthy animals were also checked. The study also evaluated the effect of Quinine-Artesunate co-administration on acetic acid induced algesia in mice; while brewer’s yeast induced pyrexia was carried out in rats. The effect of the single drug (Quinine or Artesunate) on pain and pyrexia was compared with concurrent administration of quinine and Ibuprofen (100 mg/kg) or artesunate and Ibuprofen. Results of Quinine-Artesunate co-administration did not show any significant reduction in glucose level compared to normal control, infected control, quinine and artesunate groups. The co-administration did not show any significant increase in liver enzymes compared to the other groups. This is in contrast to quinine group that showed significant increase (P<0.01) in liver enzyme (ALT) compared to normal control group of parasitized animal’s study. The quinine group also showed significant increase in AST level in healthy animals compared to Artesunate group (P<0.05). Quinine-Artesunate co-administration did not show significant difference in lipid peroxidation and antioxidant enzymes biomarkers in healthy animals compared to quinine, artesunate and the normal control groups. It showed less significant increase in lipid peroxidation (MDA) compared to normal control group (P<0.05) than quinine (P<0.01) and artesunate (P<0.01) groups. Quinine-Artesunate group also showed a lesser significant decrease (P<0.01) in antioxidant enzyme (CAT) compared to normal control group than artesunate and quinine groups (P<0.0001). Quinine-Artesunate group did not show significant difference in antioxidant enzymes compared to the infected control group. In healthy animals, Quinine-Artesunate did not show significant difference in organ body weight ratio between the groups but showed significant decrease in liver-body weight ratio in parasitized animals compared to the infected control group (P<0.0001). However, no significant difference was seen in liver-body weight ratio when it was compared to the normal control group as seen in the quinine, artesunate and infected control groups. A significant decrease was also seen in Quinine-Artesunate group compared to the quinine group (P<0.01) in the spleen-body weight ratio. No significant difference was seen in haematological parameters of healthy animals in Quinine-Artesunate group compared to other groups. The Quinine-Artesunate group did not show significant difference during concurrent administration of Ibuprofen in pain and pyrexia.


This study however, showed that Quinine and Artesunate are better in efficacy and safety when co-administered and does not reduce the efficacy of ibuprofen during pain and pyrexia.





CHAPTER ONE

INTRODUCTION




1.1              Preamble

Malaria is the leading cause of morbidity and mortality in most sub-Saharan Africa and Southeastern Asia. It is a mosquito-borne disease caused by a plasmodium parasite and transmitted by infected female anopheles mosquitoes (Malik and Khalafalla, 2004). Most of the malaria related morbidities and mortalities are caused by Plasmodium falciparum

(Soniran et al., 2012). Treatment involves the use of anti-malaria agents and supportive measures. In recent times Artemisinin derivatives have given rise to a renewed hope in fight against malaria, although resistance has been reported in places where their mono-therapy is common (WHO, 2009). Combination therapies have been recommended for the treatment of malaria to prevent early development of resistance to anti-malaria agents.


The Artemisinin based combination therapy (ACT), combining any of the Artemisinin derivatives with a longer acting anti-malaria agent in combating malaria, has brought about improved results in alleviating malaria (WHO, 2009).


Artesunate, a semi-synthetic derivative of Artemisinin used in the treatment of malaria when co-administered with Quinine (effective and longer acting anti-malaria) has shown greater efficacy than when either is used alone (Bartoloni et al., 2010). Co-administration of anti-malaria drugs with other supportive drugs like analgesics in patients with malaria to reduce pain and pyrexia may lead to drug-drug interaction resulting to synergistic....


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