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The Effects of Artemisia Derived Natural Products on Adipogenesis Florida International University FIU Digital Commons FIU Electronic Theses and Dissertations University Graduate School 2017 The ffecE ts of Artemisia Derived Natural Products on Adipogenesis Steven Abood Florida International University, [email protected] DOI: 10.25148/etd.FIDC001953 Follow this and additional works at: https://digitalcommons.fiu.edu/etd Part of the Biological Psychology Commons, Biology Commons, Cell Biology Commons, Evolution Commons, Health Psychology Commons, Neuroscience and Neurobiology Commons, and the Nutrition Commons Recommended Citation Abood, Steven, "The Effects of Artemisia Derived Natural Products on Adipogenesis" (2017). FIU Electronic Theses and Dissertations. 3383. https://digitalcommons.fiu.edu/etd/3383 This work is brought to you for free and open access by the University Graduate School at FIU Digital Commons. It has been accepted for inclusion in FIU Electronic Theses and Dissertations by an authorized administrator of FIU Digital Commons. For more information, please contact [email protected]. FLORIDA INTERNATIONAL UNIVERSITY Miami, Florida THE EFFECTS OF ARTEMISIA DERIVED NATURAL PRODUCTS ON ADIPOGENESIS A dissertation submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in BIOLOGY by Steven Abood 2017 To: Dean Michael R. Heithaus College of Arts, Sciences, and Education This dissertation, written by Steven Abood, and entitled The Effects of Artemisia Derived Natural Products on Adipogenesis, having been approved in respect to style and intellectual content, is referred to you for judgment. We have read this dissertation and recommend that it be approved. ____________________________________ Javier Francisco-Ortega ____________________________________ Juan Liuzzi ____________________________________ John Makemson ____________________________________ Jennifer Richards ____________________________________ M. Alejandro Barbieri, Major Professor Date of Defense: June 30, 2017 The dissertation of Steven Abood is approved. ____________________________________ Dean Michael R. Heithaus College of Arts, Sciences, and Education ____________________________________ Andres G. Gil Vice President for Research and Economic Development and Dean of the University Graduate School Florida International University, 2017 ii © Copyright 2017 by Steven Abood All rights reserved. iii DEDICATION This dissertation is dedicated with affection to my parents, Dr. Faye Abood and Maurice Abood, who journeyed from a Pennsylvania coal mining town and a Lebanese fishing village to offer their boundless love and support to a son that allowed him to embark on a journey of exploration to the deep reaches of science. You are my best friends and the paragon of compassionate human beings. iv ACKNOWLEDGMENTS A deep thanks to Dr. M. Alejandro Barbieri and Dr. Maria Luisa Veisaga for taking a chance on me and inviting me into your lab and lives. Thank you to all Barbieri Lab members for your help, friendship, humor, and patience. My appreciation to all my original Committee Members, Dr. Javier Francisco- Ortega, Dr. Juan Liuzzi, Dr. John Makemson, and Dr. Martin Tracey, for your time, suggestions, and guidance. Thanks to Dr. Jennifer Richards for stepping in to offer your time and insight as an expert reader of this dissertation. Thank you Dr. Luiz Lopez and your entire lab for so warmly welcoming me to Mendoza, Argentina and working with me each day on the experiments. Your lab and Mendoza will always have a special place in my heart. Thank you to all other present and past collaborators and researchers whose work made this dissertation possible. My appreciation to the entire Florida International University (FIU) Biology Department, the International Center for Tropical Botany, and all other funding sources for providing the support to make my research possible. Thank you to my entire family, including my grandparents, my brothers Michael and Brandon, their families, and their precious daughters Anna and Ruby, who will inherit and create a world beyond all our imaginings. Thank you again to my parents, Maurice and Faye Abood: through tireless years of work and 4 AM mornings at the bakery, through countless bookstore and museum trips, through your words of encouragement…without this I would not be who I am. I will continue to move forward to honor your sacrifices. v Thank you to all my early scientific mentors and role models, including Buckminister Fuller, who took the time to speak to my class when I was in elementary school, and my science teacher at Palmer Academy who wrote Longfellow’s words in my school yearbook that “The heights by great men reached and kept were not attained by sudden flight, but they, while their companions slept, were toiling upward in the night.” Thank you to Mr. Moo H. Kim, for my early training in martial arts and the profound parables that instilled within me the importance of belief in self, hard work, and perseverance in the face of adversity. Thank you Mr. Steven Alphabet, for your teachings and friendship. Your encouragement, selfless assistance, belief in me, and humor, helped me immensely during times of struggle. Thank you Mr. Alberto Fernandez and the Baragua Judo crew, for your teachings and good humor. Thank you CJ Strawn for your immense friendship and humor which makes this journey so much more enjoyable. Keep your tan sao up. Thank you to old friends James Harley Bailey, Tim Sheehan, Stewart Hendricks, and all the many others. I wish I could name you all, but you know who you are. A man with good friends is truly blessed, and I am grateful for this blessing and the many others in my life. vi ABSTRACT OF THE DISSERTATION THE EFFECTS OF ARTEMISIA DERIVED NATURAL PRODUCTS ON ADIPOGENESIS by Steven Abood Florida International University, 2017 Miami, Florida Professor M. Alejandro Barbieri, Major Professor For the first time in human history, more people worldwide suffer from obesity than are undernourished. Numerous health complications are associated with obesity including cardiovascular disease, Type 2 Diabetes, cancers of reproductive tissues, stroke, depression, anxiety disorders, and Alzheimer’s disease. A deeper understanding of the anti-adipogenic effects and mechanism of action of sesquiterpene lactones may have pharmacological import in the continuing search for therapeutic modalities to ameliorate the effects of this global obesity epidemic. Dehydroleucodine (DhL), 11,13-dihydro-dehydroleucodine (DH-DhL), and dehydroparashin-B (DhP), sesquiterpene lactones extracted from or derived from compounds extracted from Artemisia douglasiana, were investigated for their anti- adipogenic effects on 3T1-L1 preadipocytes. Dehydroleucodine inhibited the expression of C/EBP and PPARand also strongly blocked the expression of C/EBPβ, an early stage biomarker of early adipogenesis, in a concentration-dependent manner. Dehydroleucodine arrested the cell cycle at the G0/G1 phase, increased p27 and decreased both cyclins A and D and their vii partners (e.g., CDK2 and CDK4). Furthermore, DhL downregulated expression of histone demethylase JMJD2 as well as repressed the expression of histone methyltransferase MLL4, which in turn diminished the expression of C/EBPand PPARrespectively. 11,13-dihydro-dehydroleucodine blocked the accumulation of lipid droplets and inhibited the expression of PPARγ and C/EBPβ. Collectively, the results indicate that the inhibition of early stage preadipocyte differentiation by DH-DhL may be associated with cell cycle arrest at the G0/G1 phase. Dehydroparashin-B significantly decreased the accumulation of lipid content and downregulated the expression of CEBP PPARγ and CEBPα as well as FAS. Interestingly, the addition of DhP inhibited the number as well as the size of the lipid droplets during the differentiation of 3T3-L1 preadipocytes. Taken together, this data suggests that DThP has an important inhibitory effect on cellular pathways regulating adipocyte differentiation. viii TABLE OF CONTENTS CHAPTER PAGE I. INTRODUCTION............................................................................................................1 1.1. The Epidemiology of Obesity......................................................................................1 1.1.1. Measuring Obesity........................................................................................3 1.1.2. Global Changes in Obesity Rates.................................................................4 1.1.2.1. Gender and Obesity.......................................................................5 1.1.2.2. Regional Trends in Obesity Rates.................................................5 1.1.2.3. Childhood Obesity........................................................................7 1.1.3. Morbidity and Mortality of Obesity.............................................................7 1.1.3.1. General Health Effects..................................................................7 1.1.3.2. Cardiovascular Disease.................................................................9 1.1.3.3. Diabetes.........................................................................................9 1.1.3.4. Cancer...........................................................................................10 1.1.3.5. Diminished Recovery Outcomes..................................................10 1.1.3.6. Enhanced Pregnancy Risks...........................................................11 1.1.3.7. Mental Health................................................................................11
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