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Potential Enhancement of Dietary Isothiocyanates Combination on Biological Activities

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Potential Enhancement of Dietary Isothiocyanates Combination on Biological Activities University of Massachusetts Amherst ScholarWorks@UMass Amherst Doctoral Dissertations Dissertations and Theses July 2017 POTENTIAL ENHANCEMENT OF DIETARY ISOTHIOCYANATES COMBINATION ON BIOLOGICAL ACTIVITIES Kanyasiri Rakariyatham University of Massachusetts Amherst Follow this and additional works at: https://scholarworks.umass.edu/dissertations_2 Part of the Biology Commons, and the Food Science Commons Recommended Citation Rakariyatham, Kanyasiri, "POTENTIAL ENHANCEMENT OF DIETARY ISOTHIOCYANATES COMBINATION ON BIOLOGICAL ACTIVITIES" (2017). Doctoral Dissertations. 963. https://doi.org/10.7275/9959223.0 https://scholarworks.umass.edu/dissertations_2/963 This Open Access Dissertation is brought to you for free and open access by the Dissertations and Theses at ScholarWorks@UMass Amherst. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of ScholarWorks@UMass Amherst. For more information, please contact [email protected]. POTENTIAL ENHANCEMENT OF DIETARY ISOTHIOCYANATES COMBINATION ON BIOLOGICAL ACTIVITIES A Dissertation Presented by KANYASIRI RAKARIYATHAM Submitted to the Graduate School of the University of Massachusetts Amherst in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY May 2017 Food Science © Copyright by Kanyasiri Rakariyatham 2017 All Rights Reserved POTENTIAL ENHANCEMENT OF DIETARY ISOTHIOCYANATES COMBINATION ON BIOLOGICAL ACTIVITIES A Dissertation Presented by KANYASIRI RAKARIYATHAM Approved as to style and content by: _________________________________________ H. Xiao, Chair _________________________________________ G. Zhang, Member _________________________________________ R. J. Wood, Member ______________________________________ E. A. Decker, Department Head Department of Food Science DEDICATION I dedicated this work to my parents, who always support on every event. They never give up on me, and let me learn and grow beautifully. ACKNOWLEDGMENTS I have got chances to learn something new every day and the days I spent in Amherst mostly in Xiao’s lab at UMASS were very rewarding and nothing can be replaced. During my five years, here, I have met with a lot of people and I have learnt so much especially from my dissertation advisor, Dr. Hang Xiao, who spent time going over data with me, prepping me for presentations, giving me suggestions, and encouraging me to critically think throughout any problems. I am very proud to be part of his lab. I also thank Dr. Guodong Zhang, and Dr. Richard J. Wood for their precious time and advice toward this work. I greatly appreciate Dr. Eric A. Decker, head of department, and all of food science faculties, and staffs who helped and geared me up through this Ph.D. program. I thank all friends who I have met here and who I have known from before. Their friendships are invaluable. The past and present labmates had helped me a lot with things inside and outside laboratory. My housemates at 77-79 N Prospect Street have always been very supportive. We have passed through many things together. Special thanks to Gina for proofreading this dissertation. Finally, my friends in a small Thai community have made this place very lively. I am glad that we met and I enjoyed spending time with every one of them. v ABSTRACT POTENTIAL ENHANCEMENT OF DIETARY ISOTHIOCYANATES COMBINATION ON BIOLOGICAL ACTIVITIES MAY 2017 KANYASIRI RAKARIYATHAM, B.S., CHIANG MAI UNIVERSITY, THAILAND M.S., CALIFORNIA STATE UNIVERSITY FULLERTON Ph.D., UNIVERSITY OF MASSACHUSETTS AMHERST Directed by: Professor Hang Xiao Isothiocyanates (ITCs) such as allyl isothiocyanate (AIT) and sulforaphane (SFN) are well- known bioactives with wide range of beneficial properties, which may be consumed simultaneously through diets containing cruciferous vegetables. However, biological activities of ITCs in combinations had not been well defined. The present study evaluated the potential efficacy of AIT, SFN and their combinations on three important biological properties: anticancer, anti-inflammation and antioxidant. Our results showed that the combination between AIT and SFN led to a stronger growth inhibition on A549 non-small cell lung cancer cells than treatments with the individual compounds. The enhanced effect was proved to be synergistic by isobologram analysis. Flow cytometry analysis demonstrated that the combination treatment caused more extensive cell cycle arrest and cellular apoptosis in the cancer cells than the singular treatment. In addition, a synergy between AIT and SFN was also observed in their anti-cell migration. It is noteworthy that the AIT-SFN combination resulted in the production of intracellular reactive oxygen species (ROS), which might contribute to their inhibitory effects on cancer cells. In terms of anti-inflammation, the combination of AIT and SFN both pairing between themselves (AIT-SFN), and pairing with other dietary bioactives (AIT-CUR, AIT-LUT, and SFN-LUT) vi enhanced this beneficial property in comparison to a single compound utilization in lipopolysaccharide-induced RAW 264.7 macrophages. We observed dose-dependent and synergistic inhibition of pro-inflammatory molecules production such as nitric oxide, and interleukin-6. Western blotting showed corresponding information that the combined treatment reduced the expression levels of pro-inflammatory proteins and increased the expression of an antioxidative protein, which could contribute to their anti-inflammatory properties as well. In addition, pretreatment of RAW 264.7 cells with the AIT-SFN combination provided synergistic cytoprotective effects against tert-butyl hydroperoxide-induced oxidative damage by increasing antioxidant effects, decreasing cellular ROS, and increasing viability of RAW 264.7 cells. These protective properties were completed through phase 2 antioxidant and detoxification proteins, some of which had more dominant effects than the others, under a partial regulation of Nrf2, and NF-B transcription factors. Overall, this study proved a potential enhancement of dietary ITCs in combinations on biological activities, and provided information for developing functional foods for health benefits. vii TABLE OF CONTENTS Page ACKNOWLEDGMENTS ...................................................................................................................... v ABSTRACT ........................................................................................................................................ vi LIST OF TABLES .................................................................................................................................xi LIST OF FIGURES .............................................................................................................................. xii LIST OF ABBREVIATIONS ................................................................................................................ xiv CHAPTER 1. INTRODUCTION ................................................................................................................... 1 2. LITERATURE REVIEW ........................................................................................................... 5 2.1 Overview of Cellular Oxidative Stress, Inflammation and Carcinogenesis ................... 5 2.1.1 Oxidative Stress ............................................................................................ 5 2.1.2 Inflammation ................................................................................................ 6 2.1.3 Carcinogenesis .............................................................................................. 8 2.1.4 Association between Cellular Oxidative Stress, Inflammation, and Carcinogenesis ........................................................................................ 11 2.2 Possible Preventive and Therapeutic Mechanisms .................................................... 16 2.2.1 Antioxidation .............................................................................................. 16 2.2.2 Anti-Inflammation ...................................................................................... 19 2.2.3 Anticancer ................................................................................................... 21 2.3 Dietary Bioactive Components Against Oxidative Stress, Inflammation and Cancer .................................................................................................................. 24 2.3.1 Roles of Some Phytochemicals ................................................................... 24 2.3.1.1 Isothiocyanates ........................................................................... 24 2.3.1.1.1 Sulforaphane ............................................................... 26 2.3.1.1.2 Allyl Isothiocyanate ..................................................... 30 2.3.1.2 Luteolin ....................................................................................... 32 2.3.1.3 Curcumin ..................................................................................... 35 2.3.2 Combination of Dietary Bioactive Components as a Strategic Solution .................................................................................................. 38 3. SYNERGISTIC CHEMOPREVENTIVE EFFECT OF ALLYL ISOTHIOCYANATE AND SULFORAPHANE ON A549 NON-SMALL CELL LUNG CARCINOMA CELLS.......................... 44 viii 3.1 Introduction ................................................................................................................ 44 3.2 Materials and Methods
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