Immunomodulatory Effects of Traditional Chinese Herbal Formulation, Ginseng and Dang Gui Ten Combination (PS10) Thesis submitted for the Degree of Master of Science Michael Thomsen N.D., Dip. Bot.Med. Graduate School of Integrative Medicine Swinburne University of Technology 2006 Acknowledgements I wish to sincerely thank my supervisors at the Graduate School of Integrative Medicine at Swinburne University of Technology, Dr Luis Vitetta and head of the school, Prof Avni Sali. I would also like to thank Marilyn Johnson, for without her help I would still have been applying for ethics approval. This study would not have been possible without the support and guidance of the Graduate School of Integrative Medicine. This study would not have been possible without the enormous help I received from Dr Graham Flannery and his team of the Department of Genetics, Faculty of Science, Technology and Engineering, La Trobe University, Bundoora, Victoria. Dr Flannery has helped to pioneer the particular NK cell cytotoxicity test used in the present study. In particular I wish to thank his assistants Rosalia Bruzzese and Maria Mylonas for performing the NK cell assay. In addition, I would like to thank Dr Hijikata from Japan for her assistance in supplying Japanese research papers and sharing her clinical experience in the use of the herbal formulation that was the subject of the present study. I would further like to thank Phytamedica for manufacturing the test medication and to Analytica Laboratory for help with the analytical analysis of the ingredients and the final formulation. Lastly I would like to thank my partner and fellow herbalist, Erin Collins and my children for putting up with me while producing this thesis. i Table of Content ACKNOWLEDGEMENTS ............................................................................................................I TABLE OF CONTENT ................................................................................................................II INDEX OF TABLES ..................................................................................................................... V INDEX OF FIGURES ..................................................................................................................VI ABBREVIATIONS AND DEFINITIONS................................................................................ VII ABBREVIATIONS ......................................................................................................................... VII DEFINITIONS ..............................................................................................................................VIII ABSTRACT ................................................................................................................................ XII HERBAL PREPARATION – GINSENG & DANG GUI TEN COMBINATION (PS10) AND IMMUNE RESPONSE IN HUMANS................................................................................................................ XII STATEMENT OF ORIGINALITY .........................................................................................XIV 1. INTRODUCTION................................................................................................................. 1 1.1 OBJECTIVE OF PRESENT THESIS....................................................................................... 1 1.2 PERSPECTIVE ON TRADITIONAL MEDICINE..................................................................... 2 1.2.1 Herbal medicine used worldwide.............................................................................. 3 1.3 FORMULA DEVELOPMENT IN TRADITIONAL CHINESE MEDICINE .................................... 4 1.3.1 Hierarchy of Ingredients........................................................................................... 4 1.3.2 Formula Modification............................................................................................... 5 1.4 GINSENG AND DANG GUI TEN COMBINATION................................................................ 5 1.4.1 Analysis of the Formulation...................................................................................... 7 1.4.2 Traditional Preparation............................................................................................ 8 1.4.3 Traditional Indications ............................................................................................. 8 1.5 BIOLOGICAL CANCER THERAPY..................................................................................... 9 1.5.1 Biological Response Modifiers ................................................................................. 9 1.5.2 Herbal Biological Response Modifiers (BRM) ....................................................... 11 1.5.3 Ginseng and Dang Gui Ten Combination– a herbal BRM ..................................... 11 1.6 SCIENTIFIC INVESTIGATION.......................................................................................... 13 1.6.1 Introduction ............................................................................................................ 13 1.6.2 Immunomodulatory activities.................................................................................. 13 1.6.3 Clinical Studies....................................................................................................... 27 1.6.4 Active Constituents in Ginseng & Dang Gui Ten Combination.............................. 32 1.6.5 Toxicology Studies .................................................................................................. 35 1.7 MONOGRAPHS OF INDIVIDUAL HERBS........................................................................... 36 1.7.1 Angelica sinensis..................................................................................................... 36 1.7.2 Astragalus membranaceus...................................................................................... 42 ii 1.7.3 Atractylodes macrocephala..................................................................................... 46 1.7.4 Cinnamomum cassia ............................................................................................... 48 1.7.5 Glycyrrhiza glabra.................................................................................................. 50 1.7.6 Ligusticum wallichii................................................................................................ 56 1.7.7 Paeonia lactiflora ................................................................................................... 59 1.7.8 Panax ginseng......................................................................................................... 62 1.7.9 Poria cocos ............................................................................................................. 84 1.7.10 Rehmannia glutinosa.......................................................................................... 88 2 MATERIALS AND METHODS........................................................................................ 95 2.1 TEST MEDICATION ........................................................................................................ 95 2.2 CHEMICAL ANALYSIS ................................................................................................... 97 2.2.1 HPTLC (High Performance Thin Layer Chromatography).................................... 98 2.2.2 HPLC (High Performance Liquid Chromatography) ............................................. 98 2.2.3 Microbiology........................................................................................................... 98 2.3 OVERALL STUDY DESIGN AND OBJECTIVES ................................................................. 98 2.3.1 Study Design ........................................................................................................... 98 2.3.2 Study Population and Size....................................................................................... 99 2.3.3 Sample Selection ..................................................................................................... 99 2.3.4 Selection Bias.......................................................................................................... 99 2.3.5 Inclusions and Exclusions....................................................................................... 99 2.3.6 Compliance ............................................................................................................. 99 2.3.7 Health Status......................................................................................................... 100 2.3.8 Recent Medication Use ......................................................................................... 100 2.3.9 Test Medication..................................................................................................... 100 2.3.10 Blood Collection............................................................................................... 100 2.4 ASSAYS ...................................................................................................................... 101 2.4.1 Natural Killer Cell Cytolytic Assay....................................................................... 101 2.4.2 Lymphocyte Assay................................................................................................. 105 2.4.3 Liver function test.................................................................................................. 107 2.5 LYTIC UNIT ...............................................................................................................