EFFECT OF SUPPLEMENTATION WITH PUMPKIN SEED OIL VERSUS PUMPKIN SEEDS ON BLOOD PRESSURE AND MENOPAUSAL SYMPTOMS IN NON-HYPERTENSIVE POSTMENOPAUSAL WOMEN A DISSERTATION SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN THE GRADUATE SCHOOL OF THE TEXAS WOMAN’S UNIVERSITY DEPARTMENT OF NUTRITION AND FOOD SCIENCES COLLEGE OF HEALTH SCIENCES BY MADHURA MAIYA, M.S. DENTON, TEXAS DECEMBER 2017 Copyright © 2017 by Madhura Maiya ACKNOWLEDGMENTS There are many individuals I would like to acknowledge for their valuable contribution to my dissertation. I am incalculably grateful to my advisor and committee chair Dr. Carolyn Moore for her constant support and guidance throughout this research study and doctoral program. Dr. Moore has always come through for me both in professional and personal capacity. I could not have found a better teacher and a champion than Dr. Moore in my journey of pursuing the doctoral degree. I would like to thank my former advisor, Dr. John Radcliffe, for introducing me to this topic. I would like thank Dr. Shane Broughton for all his inputs which helped strengthen my research. I am grateful to all my committee members, Dr. Parakat Vijayagopal, Dr. Cynthia Warren, Dr. Sandra Cesario, and Dr. Rachelle Nurse for their guidance, support, and time. Every study is born out of a question, and the pursuit of this question brings in a lot of data. I could not have analyzed the data without Dr. Wanyi Wang’s statistical guidance. I am grateful for her patience in answering all my questions providing timely feedback. I am thankful to Dr. Mindy Maziarz for her guidance in analyzing blood samples for this study. I am immensely thankful to Ms. Laura Huntsberry, administrative assistant for department of Nutrition, Houston. Laura not only took care of academic scheduling but made sure I had all supplies needed for my research. I would also like to thank Dr. Lenora McWilliams, who trained me to become a phlebotomist. ii Although not an integral part of my study, there were a few people who stood by me and supported my research and without their support this journey would have taken twice as long and twice as hard. A few of them are from my Office of Research team, Dr. Donna Scott Tilley, Ms.Tracy Lindsay, Dr. Rene Paulson, and all other ORSP team members. I will always be thankful for their patience and understanding. This study would not have been possible without my subjects. I am indebted to them for not just signing up, but also for their patience in doing all that I asked them to do. Their kindness in supporting my research is unparalleled. Although I cannot name each and every friend and extended family member who has wished me well in this journey, I would like to extend my heartfelt thanks to every one of them. And given a chance I hope to return the favor and be there in their time of need. My family has been on this journey with me for many years. They have felt every one of my successes and setbacks as their own. There are not enough words to completely express my gratitude for their presence in my life. My parents, sister, parents- in-law, and brother-in-law, have unwaveringly supported me emotionally and intellectually. My husband has been my pillar and without him this pursuit would still be a dream. Finally, my babies Krisha and Viyat, who have filled my heart with pure joy and have given me strength to move ahead. I humbly thank everyone and hope I have made each one of them proud. iii Last but not least, I thank God for making me capable enough to pursue this dream and holding me steady at times when things looked bleak. I know I have been blessed to be a part of my family and an institution like Texas Woman’s University. iv ABSTRACT MADHURA MAIYA EFFECT OF SUPPLEMENTATION WITH PUMPKIN SEED OIL VERSUS PUMPKIN SEEDS ON BLOOD PRESSURE AND MENOPAUSAL SYMPTOMS IN NON-HYPERTENSIVE POSTMENOPAUSAL WOMEN DECEMBER 2017 Menopausal transition is a complex process often accompanied by a series of physiological changes that can affect overall quality of life in women. Estrogen deficiency is associated with menopausal symptoms, obesity, heart disease, osteoporosis, and diabetes. Pumpkin seeds are a good source of phytoestrogens and may exert weak estrogenic or anti-estrogenic effects. This randomized trial aimed to compare the effect of supplementation of pumpkin seeds (1½ teaspoons/ 4.1 grams a day) versus pumpkin seed oil (PSO) (2g/day) for 12 weeks on blood pressure (BP; systolic and diastolic), endothelial function, plasma lipids, C- rective protein (CRP) concentrations, and menopausal symptoms in non-hypertensive postmenopausal women. Postmenopausal women (n=27) were randomly assigned to receive pumpkin seeds or PSO for 12 weeks. BP, plasma lipids, endothelial function, and CRP concentrations were measured at baseline and 12 weeks after supplementation. Participants also completed a menopausal symptom questionnaire at baseline and after 12 weeks. Statistical analysis was performed using Statistical Package for the Social Science (SPSS-version 24). Both v within-subjects and between-subject effects were compared using repeated measures ANOVA. An alpha of 0.05 was set for significance. In the pumpkin seed group, systolic BP significantly decreased 3% (117.86±11.60 mmHg versus 114.43±14.39 mmHg; p=0.014) after supplementation compared to the PSO group. Diastolic BP at 12 weeks also decreased significantly (3%) in women belonging to the PSO group (74.85±8.61 mmHg versus 72.46±8.89 mmHg; p = 0.026) and decreased 4.5% in the pumpkin seed group (73.93±6.49 mmHg versus 70.57±7.75 mmHg; p = 0.002) from baseline. An improvement in endothelial function was seen by a non-significant increase in reactive hyperemia index (4%) and a decrease in augmentation index (25%) at 12 weeks in the pumpkin seed group. For women in the PSO group mean overall menopausal symptom score significantly decreased from baseline (15.77±6.77) to 12 weeks (8.46±5.98; p < 0.01) with a significant decrease in severity of hot-flushes (p = 0.02). There were no significant changes in plasma lipids and CRP-concentration in both groups. Pumpkin seeds exhibit cardio-protective benefits by reducing BP and a marginal improvement of endothelial function in postmenopausal women. Further studies at different doses of pumpkin seeds are required to better understand overall cardiovascular health benefits. vi LIST OF ABBREVIATIONS ACEI – angiotensin converting enzyme inhibitor AHA – American Heart Association AI – augmentation index ARB – angiotensin receptor blocker BB – beta blocker BMI – body mass index BP – blood pressure CCB – calcium channel blocker CEE – conjugated equine estrogens CHD – coronary heart disease CRP – C-reactive protein CVD – cardiovascular disease DBP – diastolic blood pressure DM – diabetes mellitus DNA – deoxyribonucleic acid ED – enterodial EE – energy expenditure EL – enterolactone eNOS – endothelial nitric oxide synthase ER – estrogen receptor ERE – estrogen response elements vii ET – estrogen therapy FFQ – food frequency questionnaire FMD – flow-mediated dilatation GPER – G protein-coupled estrogen receptor HDL – high density lipoprotein HERS – Heart and Estrogen/Progestin Replacement Study HRT – hormone replacement therapy HTN – hypertension KEEPS – Kronos Early Estrogen Prevention Study LDL – low density lipoprotein mER – membrane-associated estrogen receptors MET – metabolic equivalent of task METs – metabolic syndrome MPA – medroxyprogesterone acetate NADPH – nicotinamide adenine dinucleotide phosphate NAMS – North American Menopause Society NHANES – National Health and Nutrition Examination Surveys NO – nitric oxide PAT – peripheral arterial tone PPAR – peroxisome proliferator-activated receptor PS – pumpkin seed PSO – pumpkin seed oil viii RAS – renin angiotensin system RCT – randomized controlled trial RDA – Recommended Daily Allowance RHI – reactive hyperemia index ROS – reactive oxygen species SBP – systolic blood pressure SDG – secoisolariciresional diglucoside SECO – secoisolariciresional SWAN – Study of Women’s Health Across the Nation TC – total cholesterol TG – triglyceride US – United States VMS – vasomotor symptoms WHI – Women’s Health Initiative Study WHO – World Health Organization ix TABLE OF CONTENTS ................................................................................................................................. Page ACKNOWLEDGMENTS ........................................................................................... ii ABSTRACT ................................................................................................................. v LIST OF ABBREVIATIONS .................................................................................... vii LIST OF TABLES ..................................................................................................... xii LIST OF FIGURES .................................................................................................. xiii Chapter I. INTRODUCTION .................................................................................................... 1 II. REVIEW OF LITERATURE ................................................................................. 5 Menopause ....................................................................................................... 5 Menopausal Symptoms ........................................................................ 6 Estrogen and Heart Health ................................................................... 7 Cardiovascular health in postmenopausal