Dietary Effect of Squalene and Farnesol on the Lipid Metabolism of Obese/diabetes KK-Ay Mice and Wild-type Title C57BL/6J Mice Author(s) 劉, 少凱 Citation 北海道大学. 博士(水産科学) 甲第13538号 Issue Date 2019-03-25 DOI 10.14943/doctoral.k13538 Doc URL http://hdl.handle.net/2115/74280 Type theses (doctoral) File Information Shaokai_Liu.pdf Instructions for use Hokkaido University Collection of Scholarly and Academic Papers : HUSCAP Dietary Effect of Squalene and Farnesol on the Lipid Metabolism of Obese/diabetes KK-Ay Mice and Wild-type C57BL/6J Mice （肥満/糖尿病 KK-Ay マウスと野生型 C57BL/6J マウスの脂質代謝 に及ぼすスクワレンとファルネソールの効果） 北海道大学大学院水産科学院 海洋応用生命科学専攻 Graduate School of Fisheries Sciences Division of Marine Life Science 劉 少凱 Shaokai Liu 2019（平成 31 年） Acknowledgement My deepest gratitude goes first and foremost to Professor Kazuo Miyashita, my supervisor, for his constant encouragement and guidance. He has walked me through all the stages of the writing of this thesis. Without his consistent and illuminating instruction, this thesis could not have reached its present form. Second, I would like to express my heartfelt gratitude to Professor Masashi Hosokawa, who has instructed me the animal experiment in the past three years. I am also greatly indebted to the teachers, affairs staff and students of our laboratory who helped me a lot in the past three years, not only with the experiment, but also with the life in Hakodate. I would also like to thank Ministry of Education, Culture, Sports, Science and Technology for their generous scholarship. Without their generous support, I would not have harvested so much in Hokkaido University. I should like to express my gratitude to Makiko Muraki who always treated me kindly and helped me a lot with student affairs. Last my thanks would go to my beloved family for their loving considerations and great confidence in me all through these years. I also owe my sincere gratitude to my friends who gave me their help and time in listening to me and helping me work out my problems during the difficult course of the thesis. 皆様、ありがとうございました。 Shaokai Liu 2019.01.15 Content List of abbreviations and acronyms ................................................................. 1 General introduction ............................................................................................. 4 Chapter 1 Dietary Effect of Squalene on the Lipid Metabolism of Obese/diabetes KK-Ay Mice .............................................................................. 11 1.1 Introduction ............................................................................................................ 12 1.2 Material and Methods ............................................................................................ 14 1.2.1 Material ........................................................................................................ 14 1.2.2 Animals and diets ......................................................................................... 15 1.2.3 Ethics............................................................................................................ 16 1.2.4 Sample collection ......................................................................................... 16 1.2.5 Blood lipid analysis...................................................................................... 16 1.2.6 Hepatic lipid analysis ................................................................................... 17 1.2.7 Quantitative real-time PCR .......................................................................... 17 1.2.8 HMG-CoA reductase analysis ..................................................................... 18 1.2.9 Statistic analysis ........................................................................................... 19 1.3 Results .................................................................................................................... 19 1.3.1 Initial weight, weight gain, final body weight on day 28 and water intake. 19 1.3.2 Blood glucose levels and plasma lipid parameters. ..................................... 19 1.3.3 Tissue weights .............................................................................................. 20 1.3.4 Hepatic lipid parameters .............................................................................. 20 1.3.5 FA levels of liver lipids ................................................................................ 20 1.3.6 Gene expressions related to lipid metabolism and HMG-CoA reductase activity................................................................................................................... 21 1.4 Discussion .............................................................................................................. 45 Chapter 2 Dietary Effect of Squalene on the Lipid Metabolism of C57BL/6J Mice ...................................................................................................... 48 2.1 Introduction ............................................................................................................ 49 2.2 Material and Methods ............................................................................................ 50 2.2.1 Material ........................................................................................................ 50 2.2.2 Animals and diets ......................................................................................... 50 2.2.3 Ethics............................................................................................................ 50 2.2.4 Sample collection ......................................................................................... 51 2.2.5 Blood lipid analysis...................................................................................... 51 2.2.6 Hepatic lipid analysis ................................................................................... 51 2.2.7 Quantitative real-time PCR .......................................................................... 51 2.2.8 HMG-CoA reductase analysis ..................................................................... 52 2.2.9 Statistic analysis ........................................................................................... 52 2.3 Results .................................................................................................................... 52 2.3.1 Initial weight, weight gain, final body weight on day 28 and water intake . 52 2.3.2 Blood glucose levels and plasma lipid parameters ...................................... 52 2.3.3 Tissue weights .............................................................................................. 53 2.3.4 Hepatic lipid parameters .............................................................................. 53 2.3.5 FA levels of liver lipids ................................................................................ 53 2.3.6 Gene expressions related to lipid metabolism and HMG-CoA reductase activity................................................................................................................... 54 2.4 Discussion .............................................................................................................. 75 Chapter 3 Dietary Effect of Farnesol and Sphingomyelin on the Lipid Metabolism of Obese/diabetes KK-Ay Mice ................................................ 78 3.1 Introduction ............................................................................................................ 79 3.2 Material and Methods ............................................................................................ 80 3.2.1 Material ........................................................................................................ 80 3.2.2 Animals and diets ......................................................................................... 82 3.2.3 Ethics............................................................................................................ 83 3.2.4 Sample collection ......................................................................................... 83 3.2.5 Blood lipid analysis...................................................................................... 83 3.2.6 Hepatic lipid analysis ................................................................................... 84 3.2.7 Brain lipid analysis ...................................................................................... 84 3.2.8 Kidney lipid analysis.................................................................................... 84 3.2.9 Feces lipid analysis ...................................................................................... 84 3.2.10 Quantitative real-time PCR ........................................................................ 84 3.2.11 Endogenous antioxidant status ................................................................... 85 3.2.12 Statistic analysis ......................................................................................... 85 3.3 Results .................................................................................................................... 86 3.3.1 Initial weight, weight gain, final body weight on day 28 and water intake . 86 3.3.2 Plasma lipid parameters ............................................................................... 86 3.3.3 FA levels of plasma TL ................................................................................ 86 3.3.4 Tissue weights .............................................................................................