The Effects of Supplementing with Constituents of Flaxseed during Exercise Training on Inflammation in Older Adults A Thesis Submitted to the College of Graduate Studies and Research in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in the College of Kinesiology University of Saskatchewan Saskatoon By Stephen Mark Cornish © Copyright Stephen Mark Cornish, June 2008. All rights reserved. Permission to Use In presenting this thesis/dissertation in partial fulfillment of the requirements for a Postgraduate degree from the University of Saskatchewan, I agree that the Libraries of this University may make it freely available for inspection. I further agree that permission for copying of this thesis/dissertation in any manner, in whole or in part, for scholarly purposes may be granted by the professor or professors who supervised my thesis/dissertation work or, in their absence, by the Head of the Department or the Dean of the College in which my thesis work was done. It is understood that any copying or publication or use of this thesis/dissertation or parts thereof for financial gain shall not be allowed without my written permission. It is also understood that due recognition shall be given to me and to the University of Saskatchewan in any scholarly use which may be made of any material in my thesis/dissertation. Requests for permission to copy or to make other uses of materials in this thesis/dissertation in whole or part should be addressed to: Dean College of Kinesiology University of Saskatchewan 87 Campus Drive Saskatoon, Saskatchewan S7N 5B2 Canada OR Dean College of Graduate Studies and Research University of Saskatchewan 107 Administration Place Saskatoon, Saskatchewan S7N 5A2 Canada i Abstract This thesis evaluated supplementation with two components of flaxseed during exercise training on inflammation in older adults. Experiment 1: This experiment assessed secoisolariciresinol diglucoside (SDG) supplementation during aerobic exercise training on inflammation in older adults. Methods: One hundred subjects aged 50y or older were randomized to receive either SDG or placebo before completing a 6-month walking program. Fasting concentrations of interleukin-6 and tumor necrosis factor-α, glucose, triacylglycerol (TAG), high density lipoprotein (HDL), low density lipoprotein, and total cholesterol as well as leukocyte cell count were measured every two months while body composition, resting blood pressure, and a composite Z-score of six metabolic syndrome risk factors were assessed at baseline and 6 months. Results: Men on placebo increased metabolic syndrome composite Z-score (p<0.05). TAG increased (p=0.017) in men on placebo relative to men on SDG and men on SDG decreased (p=0.045) DBP relative to men on placebo. Conclusions: SDG had no effect on inflammation while it is effective in attenuating risk factors associated with metabolic syndrome in older males but not females. Experiment 2: This experiment evaluated alpha-linolenic acid (ALA) supplementation during strength exercise training on inflammation in older adults. Methods: Fifty-one healthy older adults (65.4±0.8y) were randomized to receive ALA or a placebo before completing a 12 wk strength training program. Subjects were evaluated at baseline and 12 weeks for TNF-α and IL- 6, muscle strength, body composition, and muscle thickness. Results: Males supplementing with ALA decreased IL-6 concentration (p=0.003). The female placebo and male ALA group had a significant increase in knee flexor thickness (p<0.05). Chest and leg press strength, lean tissue mass, and muscle thickness significantly increased, while percent fat and total body mass ii decreased with training (p<0.05), with no difference between ALA and placebo. Conclusions: ALA lowers IL-6 in older men, but has minimal effect on muscle mass and strength during resistance training. General Conclusion: A composite score of metabolic syndrome is attenuated in males supplementing with SDG. ALA reduces IL-6 in older men. Older men, but not older women, derive specific health benefits from increased consumption of components of flaxseed consumed during an exercise program. iii Acknowledgements This thesis would not have been possible without the help and assistance of many individuals. First, I would like to thank my advisor, Dr. Philip Chilibeck. Without your time, guidance, support and friendship over the past four years this thesis would not have come to fruition. I would especially like to thank-you for hiring my wife, Denise, as a research assistant. This went a long way to keeping our family‘s needs met and it is appreciated very much. Also, I would like to acknowledge the members of my committee: Dr. Jane Alcorn, Dr. Adam Baxter- Jones, Dr. Karen Chad, and Dr. Robert Faulkner. Your collective insight and critical reading of this thesis is appreciated and I value the time, knowledge and wisdom each of you shared with me while obtaining this degree. Also, thank you to Dr. Russ Hepple for committing his time and expertise in acting as my external examiner. I also wish to acknowledge the individuals who aided in exercise programming and helping with data collection including: Nathan Jantz, Charlene Magnus, and Shane Schwanbeck. Also, I appreciate the efforts of Betty Giesbrecht in collecting blood samples. Further, Dr. Vijitha Senajake and Dr. Tahereh Talaei Khozani were of immense help in guiding me through the procedures for assaying the blood samples from these studies and I appreciate that Dr. Bernard Juurlink was willing to let them dedicate a significant portion of time in helping me when they had many things to do in his lab as well. The financial support of the Saskatchewan Flax Development Commission and the College of Kinesiology is greatly appreciated as it aided in completing my Ph.D. studies. Also, the financial support of the Heart and Stroke Foundation of Saskatchewan and the Gatorade Sport Science Institute is acknowledged as funding for the two studies was provided by these iv agencies. I would also like to thank Archer Daniels Midland and WN Pharmaceuticals for the product support they provided for the research projects of this thesis. Finally, I would like to publically acknowledge the support of my family while ‗back in school‘. Thank you Denise for agreeing to let me pursue my dream and for loving me through ‗thick‘ and ‗thin‘; this thesis would not have happened if I did not have your encouragement, patience, and understanding the past four years of our lives together. Thank you Jonah for being patient with Daddy when he was ―always working on his computer‖ and for being a son who is full of life and always willing to love your Dad despite his many shortcomings. Levi James, you have brought much joy into my life, I hope for all the best for you in the future and I will strive to give you a great life as your parent. v Dedication This thesis is dedicated to my late grandparents: Lyla Myrtle (Aspinall) Taylor (August 15, 1927-June 16, 2000), Donald Thomas Taylor (June 8, 1925-June 13, 1996), Ellen Ruth (Langman) Cornish (July 27, 1917-February 2, 1999), and Lloyd George Cornish (April 22, 1917-March 10, 2002). Each of these wonderful people passed from this world to the next as a result of diseases related to inflammation. It is my hope and desire to continue researching how exercise and nutrition will reduce the inflammatory process associated with many types of disease so to reduce suffering and pain in old age; I want people to age successfully! I can only hope that the suffering and pain my grandparents experienced due to inflammatory disease can and will be alleviated or prevented in older adults as a result of my chosen career. Deuteronomy 28 discusses the blessings and curses of God on the nation of Israel and says: ―If you fully obey the LORD your God and carefully follow all his commands I give you today, the LORD your God will set you high above all the nations on earth … However, if you do not obey the LORD your God and do not carefully follow all his commands and decrees I am giving you today, all these curses will come upon you and overtake you: … The LORD will plague you with diseases until he has destroyed you from the land you are entering to possess. The LORD will strike you with wasting disease, with fever and inflammation, with scorching heat and drought, with blight and mildew, which will plague you until you perish.‖ I wonder how much of this is applicable to our society today. I am grateful that in the New Testament book of Revelation an answer to the problem is present: “And I heard a loud voice from the throne saying, ‗Now the dwelling of God is with men, and he will live with them. They will be his people, and God himself will be with them and be their God. He will wipe every tear from their eyes. There will be no more death or mourning or crying or pain, for the old order of things has passed away.‘‖ May we all be able to persevere and ease the suffering, pain, and disease experienced on this earth until then. vi Table of Contents Permission to Use i Abstract ii Acknowledgements iv Dedication vi Table of Contents vii List of Tables xi List of Figures xiii List of Abbreviations xv Chapter 1: General Introduction and Review of Literature 1 1.1 General Introduction 1 1.2 Overview of the Immune System 4 1.2.1 Myeloid Immune Cells 6 1.2.2 Lymphoid Immune Cells 7 1.2.3 Soluble Mediators of the Immune System 9 1.2.3.1 Cytokines 9 1.2.4 Inflammatory Response 16 1.3 Review of Literature 19 1.3.1 Aging and Inflammation 19 1.3.2 Cytokines