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Interventionsstudie Zur Rolle Von Isoflavonen Im Fettstoffwechsel Gesunder Postmenopausaler Frauen

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Interventionsstudie Zur Rolle Von Isoflavonen Im Fettstoffwechsel Gesunder Postmenopausaler Frauen Max Rubner-Institut Bundesforschungsinstitut für Ernährung und Lebensmittel Institut für Physiologie und Biochemie der Ernährung Interventionsstudie zur Rolle von Isoflavonen im Fettstoffwechsel gesunder postmenopausaler Frauen Dissertation zur Erlangung des Grades Doktorin der Ernährungs- und Lebensmittelwissenschaften (Dr. troph.) der Landwirtschaftlichen Fakultät der Rheinischen Friedrich-Wilhelms-Universität Bonn von Ann Katrin Engelbert aus Gummersbach Bonn 2018 Referent: Professor Dr. Achim Bub Korreferent: Professor Dr. Peter Stehle Tag der mündlichen Prüfung: 18.01.2018 Angefertigt mit Genehmigung der Landwirtschaftlichen Fakultät der Universität Bonn Teile dieser Arbeit wurden vorab veröffentlicht in: A.K. Engelbert, S.T. Soukup, A. Roth, N. Hoffmann, D. Graf, B. Watzl, S.E. Kulling, A. Bub, Isoflavone supplementation in postmenopausal women does not affect leukocyte LDL receptor and scavenger receptor CD36 expression: A double-blind, randomized, placebo-controlled trial, Mol Nutr Food Res. 2016; 60(9):2008-19. Danksagung Ich danke Professor Dr. Achim Bub ganz herzlich für die Bereitstellung des Themas, seine kompetente fachliche Betreuung, Diskussionsbereitschaft und Unterstützung in den letzten Jahren. Professor Dr. Stehle möchte ich für die freundliche und unkomplizierte Übernahme der Korreferententätigkeit herzlich danken. Martina Broßart, Claudia Dörr, Tanja Gadau, Susanne Gräber, Eva Hoch, Anita Kriebel, Ute Stadler- Prayle und Ulrike Rossi, meinen Kolleginnen des Studienzentrums, gilt mein besonderer Dank für ihre Hilfe und Unterstützung vor, während und nach der Studie. Dr. med. Claudia Dörr gilt mein besonderes Dankeschön für die Durchführung der Fettbiopsien. Zudem geht mein Dank an Lars Uhlmann für seine hilfreichen Tipps hinsichtlich der Durchflusszytometrie. Besonderer Dank gebührt ebenfalls Dr. Alex Roth und Dr. Paola Ferrario für ihre statistische Fachkenntnis und insbesondere Paola für die konstruktiven Diskussionen und das gewissenhafte Korrekturlesen. Dr. Sebastian Soukup möchte ich herzlich für die Isoflavonanalytik im Extrakt, Plasma und Fettgewebe danken. Bjørk Ditlev Larsen, der die Whole Genome Sequenzierung der Biopsieproben an der University of Southern Denmark durchgeführt hat, möchte ich ebenfalls meinen Dank aussprechen. Und auch Dr. Andreas Dötsch danke ich sehr für seine hilfreichen Tipps hinsichtlich der Auswertung der Genexpressionsdaten. Dr. Daniela Graf danke ich dafür, dass sie mich in die Welt der Durchflusszytometrie eingeführt hat und für jeden Tipp, jede Anmerkung, Diskussion und Aufmunterung in den letzten Jahren. Dani, vielen Dank für alles! Bei meinen Masterandinnen Nadine Hoffmann, Kira Dölp und Rebecca Veit möchte ich mich sehr für ihre Mitarbeit während der Studie bedanken. Bei Daniela Graf, Christiane Mulsow, Andrea Gille, Benedikt Merz und ganz besonders Nathalie Siegel, möchte ich mich einerseits für die zahlreichen fachlichen Diskussionen und andererseits auch für die mal weniger fachlichen Gespräche bedanken. Es macht Spaß mit Menschen wie euch zusammenarbeiten zu dürfen. Fürs Korrekturlesen möchte ich Dr. Stefanie Herberth und Sarah Schmidt meinen besonderen Dank aussprechen. Meiner Familie möchte ich von Herzen für ihre immerwährende Unterstützung danken. Inhaltsverzeichnis 1 Einleitung ........................................................................................................................ 1 1.1 Menopause....................................................................................................................... 2 1.2 Lipidstoffwechsel ............................................................................................................ 4 1.2.1 Lipoproteine .................................................................................................................... 4 1.2.1.1 Lipoproteinmetabolismus .............................................................................................. 5 1.2.1.2 Exogener Lipoproteinmetabolismus .............................................................................. 5 1.2.1.3 Endogener Lipoproteinmetabolismus ............................................................................ 6 1.2.1.4 Reverser Cholesteroltransport ....................................................................................... 6 1.2.2 LDL-Rezeptor ................................................................................................................. 7 1.2.2.1 Feedbackregulation der Cholesterolhomöostase ........................................................... 8 1.2.2.2 LDLR-Expression auf mononukleären Zellen des peripheren Blutes ........................... 9 1.2.2.3 LDLR-Expression auf Adipozyten ................................................................................ 9 1.2.3 Scavenger Rezeptor CD36 .............................................................................................. 9 1.2.3.1 CD36-Expression auf Monozyten und Makrophagen ................................................. 10 1.2.3.2 CD36-Expression auf Adipozyten ............................................................................... 11 1.2.4 Fettgewebe..................................................................................................................... 11 1.2.5 Einfluss des Fettgewebes und der Körperfettverteilung auf den Lipid- und Lipoproteinmetabolismus .............................................................................................. 12 1.2.6 Fettstoffwechsel in der Menopause ............................................................................... 14 1.2.6.1 Lipid- und Lipoproteinstoffwechsel in der Menopause ............................................... 14 1.2.6.2 Körperfettverteilung in der Menopause ....................................................................... 15 1.3 Isoflavone ...................................................................................................................... 17 1.3.1 Struktur, Vorkommen und Aufnahme der Isoflavone ................................................... 17 1.3.2 Bioverfügbarkeit und Metabolisierung der Isoflavone .................................................. 18 1.3.2.1 Absorption ................................................................................................................... 18 1.3.2.2 Metabolisierung ........................................................................................................... 19 1.3.3 Isoflavonwirkungen ....................................................................................................... 20 1.3.3.1 Estrogenrezeptor-vermittelte Wirkungen .................................................................... 20 i 1.3.3.2 Nicht ER-abhängige Wirkungen .................................................................................. 21 1.3.3.3 Potentielle Wirkungen von Isoflavonen bei Postmenopausalen Frauen ...................... 21 1.3.3.4 Antioxidative Kapazität der Isoflavone ....................................................................... 22 1.3.3.5 Isoflavoneffekte auf den Lipid- und Lipoproteinmetabolismus sowie die Körperzusammensetzung ............................................................................................. 23 2 Problemstellung und Ziele der Arbeit ........................................................................... 25 3 Material und Methoden ................................................................................................. 27 3.1 Studiendesign ................................................................................................................ 27 3.1.1 Probandinnen ................................................................................................................. 28 3.1.1.1 Fallzahlbestimmung ..................................................................................................... 29 3.1.2 Interventionsprodukt ..................................................................................................... 29 3.2 Geräte, Reagenzien und Verbrauchsmaterialien ........................................................... 30 3.3 Methoden ....................................................................................................................... 34 3.3.1 Anthropometrische Daten .............................................................................................. 34 3.3.2 Körperzusammensetzung .............................................................................................. 34 3.3.3 Blutproben und Analysen .............................................................................................. 35 3.3.3.1 Serum- und Plasmagewinnung .................................................................................... 36 3.3.3.2 Quantifizierung der Isoflavone im Plasma .................................................................. 36 3.3.4 Durchflusszytometrische Quantifizierung der LDLR- und CD36-Expression auf Leukozyten .................................................................................................................... 36 3.3.4.1 Lymphozytäre LDLR-Expression ............................................................................... 37 3.3.4.2 Leukozytäre CD36-Expression .................................................................................... 38 3.3.5 Biopsie des abdominellen subkutanen Fettgewebes ...................................................... 39 3.3.5.1 RNA Isolierung aus Fettgewebe .................................................................................
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