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www.defra.gov.uk Plant substances as alternatives for animal products in traditional medicines Report submitted to the Department for Environment Food and Rural Affairs September 2006 Dr Celia M Bell, Academic Group Chair, Human and Healthcare Sciences, School of Health and Social Sciences, Middlesex University, UK and Professor Monique SJ Simmonds, Head of Biological Interactions, Jodrell Laboratory, Royal Botanic Gardens (RBG), Kew, UK Researchers: Dr Sandra S Appiah, Middlesex University, UK Dr Melanie-Jayne R Howes, Jodrell Laboratories, RBG, Kew, UK Plant substances as alternatives for animal products in traditional medicines Report submitted to the Department for Environment Food and Rural Affairs September 2006 Dr Celia M Bell, Academic Group Chair, Human and Healthcare Sciences, School of Health and Social Sciences, Middlesex University, UK and Professor Monique SJ Simmonds, Royal Botanic Gardens, Kew, UK Researchers: Dr Sandra S Appiah, Middlesex University, UK Dr Melanie-Jayne R Howes, Royal Botanic Gardens, Kew, UK Department for Environment, Food and Rural Affairs Nobel House 17 Smith Square London SW1P 3JR Telephone 020 7238 6000 Website: www.defra.gov.uk © Crown copyright 2007 The text in this document (excluding the Royal Arms and departmental logos) may be reproduced free of charge in any format or medium provided that it is reproduced accurately and not used in a misleading context. The material must be acknowledged as Crown copyright and the title of the document specified. Any enquiries relating to the copyright in this document should be addressed to The Information Policy Team, Office of Public Sector Information, St Clements House, 2–16 Colegate, Norwich, NR3 1BQ. Fax: 01603 723000 or e-mail: [email protected] Product code PB 11737 We thank DEFRA and IFAW Charitable Trust for their financial support for this project. We also thank the CITES TEAM (Special Operations District) at Heathrow airport for the kind donation of the rhino horn sample, and the National Museums of Scotland for providing a sample of tiger bone. Other contributors Middlesex University, UK Dr Henry Lee Dr John Langley Dr Huw Jones Professor Mike Revitt Peter Lister (Technician) Alan LaGrue (Technician) John Schmitt (Technician) Jodrell Laboratory, Biological Interactions, RBG, Kew, UK Dr Tetsuo Kokubun Christine Leon Dr Elaine Porter Cheryl Waring Dr Renée Grayer Dr Geoffrey Kite Dr Nigel C Veitch Centre for Pharmacognosy and Phytotherapy, The School of Pharmacy, University of London, UK Prof. Michael Heinrich Dr Paul Bremner AstraZeneca, Research and Development, Charnwood, UK David Wilkinson Dr Craig Lambert Herbal Suppliers Jo Liu (Mayway Ltd, UK) Paul Skipworth (Kingham herbs and Tinctures, UK) Traditional Chinese Medicine Practitioners Dr Bing Chung Chan (Practitioner, main advisor) Professor Dang Yi (Middlesex University, UK) Dr Lin Zhixiu (Middlesex University, UK) Dr Kaicun Zhao (Middlesex University, UK) Dr Charlie Buck (Practitioner) Dr Ke Wang (Practitioner) Dr Su (Practitioner) Mrs Hong Wen Contents Contents i Figures v Tables vi Executive summary vii Section 1. Introduction and overview 1 1.1. Introduction 1 1.1.1. The popularity of traditional medicine 1 1.1.2. The threat to endangered species from their use in traditional medicine 1 1.1.3. The need for research into the use of substitutes for specimens of endangered species 2 1.1.4. Middlesex University and traditional medicine 2 1.2. The current research project 3 1.2.1. Aims of the project 3 1.3. Structure of the report 4 Section 2. Rationale for selection of herbs as potential alternatives to bear bile 5 2.1. Introduction 5 2.2. Sources of bear bile 5 2.3. Bear bile and its constituents in TCM and Western medicine 6 2.4. Bear bile in TCM 6 2.5. Selection of herbs and herbal prescriptions for investigation 6 2.5.1. Criteria used for selection of single herbs 6 2.5.2. Criteria used for selection of herbal prescriptions for study 8 2.6. Single herbs chosen as potential alternatives to bear bile in TCM: summary of reputed and pharmacological effects 9 2.6.1. Huang Qin (Radix Scutellariae) 9 2.6.2. Huang Lian (Rhizoma Coptidis) 9 2.6.3. Huang Bai (Cortex Phellodendri) 9 2.6.4. Zhi Zi (Fructus Gardeniae) 10 2.6.5. Chuan Xin Lian (Herba Andrographis) 10 2.6.6. Zhi Mu (Rhizoma Anemarrhena) 11 2.6.7. Da Huang (Radix et Rhizoma Rhei) 11 i Contents 2.7. Prescriptions chosen for investigation as potential alternatives to bear bile in TCM 11 2.7.1. Orengedokuto 11 2.7.2. Dia-Orengedokuto 12 2.8. Alternatives to bear bile: summary 13 Section 3. Rationale for selection of herbs as potential alternatives to rhino horn 14 3.1. Introduction 14 3.2. Sources of rhino horn 14 3.3. Rhino horn and its constituents 14 3.4. Rhino horn in TCM 15 3.5. Selection of herbs and prescriptions for investigation 15 3.5.1. Criteria used for selection of single herbs 15 3.5.2. Criteria used for selection of herbal prescriptions 16 3.6. Single herbs as potential alternatives to rhino horn: reputed and pharmacological effects 18 3.6.1. Ban Lan Gen (Radix Isatidis) 18 3.6.2. Chi Shao (Radix Paeoniae Rubra) 18 3.6.3. Mu Dan Pi (Cortex Moutan) 19 3.6.4. Dan Shen (Radix Salvia Miltiorrhizae) 19 3.6.5. Jin Yin Hua (Flos Lonicerae) 20 3.6.6. Lian Qiao (Fructus Forsythia) 20 3.6.7. Sheng Di Huang (Radix Rehmanniae) 20 3.6.8. Xuan Shen (Radix Scrophulariae) 21 3.6.9. Zi Cao (Radix Lithospermi Arnebaie) 21 3.7. Prescriptions traditionally containing rhino horn: reputed and pharmacological effects 21 3.7.1. Qing Ying Tang 21 3.7.2. Qingwen Baidu Yin 21 3.7.3. Xi Jiao Dihuang Tang 22 3.7.4. Sheng Xi Dan 22 3.7.5. Qing Gong Tang 22 3.7.6. Zhi Zi Jin Hua 22 3.8. Alternatives to rhino horn: summary 22 ii Contents Section 4. Rationale for selection of herbs as potential alternatives to tiger bone 23 4.1. Introduction 23 4.2. Sources of tiger bone 23 4.3. Tiger bone and its constituents 23 4.4. Tiger bone in TCM 24 4.5. Selection of herbs 24 4.5.1. Criteria used for selection of single herbs 24 4.5.2. Criteria used for the selection of prescriptions 25 4.6. Single herbs and other TCM remedies as potential alternatives to tiger bone: reputed and pharmacological effects 31 4.6.1. Bai Shao (Radix Paeoniae Alba) 31 4.6.2. Bai Zhu (Rhizoma Atractylodis Macrocephalae) 31 4.6.3. Cang Zhu (Rhizoma Atractylodis) 31 4.6.4. Chuan Xiong (Rhizoma Chuanxiong) 31 4.6.5. Dang Gui (Radix Angelicae Sinensis) 32 4.6.6. Di Huang (Radix Rehmanniae) 32 4.6.7. Du Huo (Radix Angelicae Pubescentis) 32 4.6.8. Du Zhong (Cortex Eucommiae) 33 4.6.9. Fang Feng (Radix Saposhnikoviae) 33 4.6.10. Fu Ling (Poria) 33 4.6.11. Gui Zhi (Ramulus Cinnamomi) 33 4.6.12. Ji Xue Teng (Caulis Spatholobi) 34 4.6.13. Lu Lu Tong (Fructus Liquidambaris) 34 4.6.14. Mu Gua (Fructus Chaenomelis) 34 4.6.15. Mu Xiang (Radix Aucklandiae) 34 4.6.16. Niu Xi (Radix Achyranthis Bidentatae) 35 4.6.17. Qin Jiao (Radix Gentiianae Macrophyllae) 35 4.6.18. Ren Shen (Radix Ginseng) 35 4.6.19. Rou Gui (Cortex Cinnamomi) 35 4.6.20. San Qi (Radix Notoginseng) 36 4.6.21. Sang Zhi (Ramulus Mori) 36 4.6.22. Wei Ling Xian (Radix Clematidis) 36 4.6.23. Xu Duan (Radix Dipsaci) 36 4.6.24. Yin Yang Huo (Herba Epimedii) 37 4.7. Alternatives to tiger bone: summary 37 iii Contents Section 5. Biological and chemical methods used to study plant and fungal material 38 5.1. Introduction 38 5.2. Materials 39 5.3. Authentication techniques for trade TCM material 39 5.3.1. HPLC (UV-DAD) method 40 5.3.2. LC-MS method 40 5.3.3. TD-GC-MS method 40 5.4. Methods for fractionation and isolation of compounds 41 5.4.1. Fractionations of Scutellaria baicalensis and isolation of compounds 41 5.4.2. Fractionation of Salvia miltiorrhiza and Qing Ying Tang 41 5.5. Method for anti-bacterial tests 42 5.6. NF−κΒ Studies using the IL-6 promoter assay method 43 5.7. Method for cytochrome P450 3A4 inhibition studies 43 5.7.1. CYP3A4 LC-MS method 44 5.8. Statistical analysis 44 Section 6. Results and discussion 45 6.1. Bear bile: bioassay results and discussion 45 6.1.1. Anti-bacterial tests 45 6.1.2. Anti-inflammatory (NF−κΒ) tests 46 6.1.3. Cytochrome P450 3A4 tests 49 6.1.4. Conclusions 51 6.2. Rhino horn: bioassay results and discussion 51 6.2.1. Anti-bacterial tests 51 6.2.2. Anti-inflammatory (NF−κΒ) tests 53 6.2.3. Cytochrome P450 3A4 tests 55 6.2.4. Conclusions 57 6.3. Tiger bone: bioassay results and discussion 57 6.3.1. Anti-inflammatory (NF−κΒ) tests 58 6.3.2. Cytochrome P450 3A4 tests 60 6.3.3. Conclusions 60 iv Contents Section 7. Conclusions 61 7.1. General conclusions 61 7.2. Herbs selected as potential alternatives to animal products 61 7.2.1. Herbs proposed as potential alternatives to bear bile used in TCM 61 7.2.2. Prescriptions proposed as potential alternatives to bear bile used in TCM 62 7.2.3. Herbs proposed as potential alternatives to rhino horn used in TCM 62 7.2.4.