TABLE OF CONTENTS
TITLE PAGE
CERTIFICATION
DEDICATION
ACKNOWLEDGEMENT
TABLE OF CONTENT
LIST OF TABLES
LIST OF FIGURES
ABSTRACT
CHAPTER ONE
INTRODUCTION1
1.1 AIM OF THESTUDY2
1.2 OBJECTIVES OF THESTUDY2
1.3 LIST OF ABBREVIATIONS3
CHAPTER TWO: LITERATURE REVIEW
2.1 STRESS4
2.1.1 TYPES OF STRESSORS5
2.1.2 EFFECTS OF OXIDATIVE STRESS ON FEMALE REPRODUCTIVE SYSTEM6
2.2 THE MAMALIAN FEMALE REPRODUCTIVE SYSTEM
2.2.1 FEMALE REPRODUCTIVE FUNCTIONS IN MAN7
2.2.1.1 OVARIES7
2.2.1.2 FALLOPIAN TUBES7
2.2.1.3 UTERUS 7
2.2.1.4 VAGINA7
2.2.2 FEMALE REPRODUCTIVE SYSTEM IN RATS8
2.2.2.1 ESTROUS CYCLE9
2.2.2.2 ESTROUS CYCLE PHASES9
2.2.2.3 ENVIRONMENTAL FACTORS AFFECTING THE ESTROUS CYCLE10
2.2.2.4 MATING AND REPRODUCTIVE BEHAVIOR 11
2.2.2.5 PREGNANCY DETECTION 11
2.2.2.6 FERTILIZATION AND EARLY EMBRYONIC DEVELOPMENT 11
2.2.2.7 EMBRYO IMPLANTATION12
2.2.2.8 GESTATION, PARTURITION AND WEANING13
2.2.3 HORMONAL CONTROL OF PREGNANCY IN RATS 14
2.2.3.1 HORMONAL CHANGES15
2.3 SLEEP DEPRIVATION
2.3.1 INTRODUCTION19
2.3.2 SLEEP LOSS AND ADRENOCORTICAL ACTIVITY21
2.3.3 SLEEP LOSS AND METABOLISM22
2.3.4 SLEEP DEPRIVATION AND ADVERSE MATERNAL AND FOETAL OUTCOMES24
2.3.5 MECHANISMS26
2.4 OXIDATIVE STRESS AND PREGNANCY27
2.4.1 ANTIOXIDANTS29
2.4.2 REACTIVE OXYGEN SPECIES AND THEIR PHYSIOLOGICAL ACTIONS 29
2.4.3 OXIDATIVE STRESS IN FEMALE REPRODUCTION32
2.4.4 OXIDATIVE STRESS AND PREGNANCY33
2.5 OXIDATIVE STRESS ANDINFLAMMATION33
2.5.1 C-REACTIVE PROTEIN36
2.6 GLUTATHIONE(GSH)37
2.6.1 D-RIBOSE L-CYSTEINE41
2.6.2 EFFECTS OF D-RIBOSE L-CYSTEINE ON GSH LEVEL42
CHAPTER THREE: MATERIALS AND METHODOLOGY
3.1 MATERIALS43
3.2 ANIMALS43
3.2.1 INDUCTION OF PREGNANCY44
3.3 EXPERIMENTAL DESIGN44
3.4 D-RIBOSE L-CYSTEINE PREPARATION, DOSAGE AND ADMINISTRATION 45
3.5 PARADOXICAL SLEEP DEPRIVATION PROTOCOL 45
3.6 ANIMAL EUTHANASIA, BLOOD SAMPLING AND TISSUE COLLECTION 45
3.7 DETERMINATION OF OXIDATIVE PARAMETERS47
3.7.1 DETERMINATION OF SUPEROXIDE DISMUTASE (SOD) 47
3.7.2 DETERMINATION OF CATALASE (CAT) 48
3.7.3 DETERMINATION OF GLUTATHIONE(GSH) 48
3.7.4 DETERMINATION OF MAIONDEALDEHYDE (MDA) 49
3.8 STATISTICAL ANALYSIS 49
CHAPTER FOUR: RESULT 50
CHAPTER FIVE:
5.1 DISCUSSION AND FINDINGS61
5.2 CONCLUSION62
REFERENCES
LIST OF TABLES
TABLE 1: BEHAVIOUR AND VAGINAL CYTOLOGY WITH OESTROUS CYCLE PHASES10
TABLE 2: SOURCES AND FUCTIONS OF THE HORMONES OF THE FEMALE REPRODUCTIVE SYSTEM18
TABLE 3: EFFECT OF PSD AND DRLC ADMINISTRATION ON NUMBER AND WEIGHT OF IMPLANTATION IN RATS.49
TABLE 4: EFFECT OF PSD AND DRLC ADMINISTRATION ON FETAL PARAMETERS AND PREGNANCY OUTCOME IN RATS.53
TABLE 5: OESTRADIOL CONCENTRATION55
LIST OF FIGURES
FIGURE 1: HORMONAL CONTROL OF PREGNANCY IN RATS14
FIGURE 2: EFFECTS OF OXIDATIVE STRESS ON FEMALE REPRODUCTIVE SYSTEM30
FIGURE 3: EFFECT OF PSD AND DRLC ADMINISTRATION ON SERUM CONCENTRATION OF CORTICOSTERONE IN PREGNANT RATS SACRIFICED ON DIFFERENT DAYS OF GESTATION.56
FIGURE 4: EFFECT OF PSD AND DRLCADMINISTRATION ON MDA CONCENTRATION INSERUM OF PREGNANT RATS.57
FIGURE 5: EFFECT OF PSD AND DRLC ADMINISTRATION ON LEVEL OF SOD ACTIVITY INSERUM OF PREGNANTRATS.58
FIGURE 6: EFFECT OF PSD AND DRLC ADMINISTRATION ON LEVEL OF CAT ACTIVITY INSERUM OF PREGNANT RATS.
FIGURE 7: EFFECT OF PSD AND DRLC ADMINISTRATION ON GSH CONCENTRATION INSERUM OF PREGNANT RATS.
FIGURE 8: EFFECT OF PSD AND DRLC ADMINISTRATION ON CRP CONCENTRATION INSERUM OF PREGNANT RATS.
ABSTRACT
In a healthy body, ROS (reactive oxygen species) and antioxidants remain in balance. When the balance is disrupted towards an overabundance of ROS, oxidative stress (OS) occurs. OS influences the entire reproductive lifespan of a woman and even thereafter (i.e. menopause). OS results from an imbalance between pro-oxidants (free radical species) and the body's scavenging ability (antioxidants). ROS build up in the body can be aided by stress e.g. sleep deprivation. Sleep deprivation is becoming increasingly common in modern society, especially in undergraduate students. Sleep is crucial for optimal functioning of the human body.
A modified multiple platform was used in this experiment, which uses a REM technique to manipulate sleep deprivation, to actuate sleep deprivation in pregnant rats. About 60 female Sprague-Dawley rats were used for this experiment, with 15 rats per group, 5 for each of 3 subgroups and four groups totalling to 60 rats. The rats were housed in a fully aerated standard cage at room temperature with a 12hour dark and light cycle. The rats were fed on daily basis in their respective cages with rat chow. Oestrous cycle was observed in the female rats and rats on proestrus were introduced to male rats on proestrus evening for mating. Mating was confirmed by the presence of sperm cells in the vaginal smear and was taken as day one of pregnancy. DRLC was administered to the GLU groups. A plastic cage (40 x 30 cm) with 7 columns (platforms, 5 x 3 cm) was filled with 1 cm water. Five mice, all from the same cage, were placed in each cage with water and food for twenty hours every day for seven days from the onset of pregnancy. The water is changed daily and the rats are removed from the stress cage to recuperate for four hours.The loss of muscle tone associated with sleep deprivation caused them to touch the water and wake up.
The result shows significant decrease in the number of foetusat term in the PSD group when compared to the control group (P
