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NUS Corporate Presentation Slides Updated 4 March 2010 Better Foods for Better Health, 3rd edition 12 – 14 September 2012 Antioxidants, Nutrition and Ageing – Miracles, Myths and Misunderstandings Professor Barry Halliwell Tan Chin Tuan Centennial Professor Deputy President (Research & Technology) National University of Singapore Antioxidants, Nutrition and Ageing Current Research in the Halliwell Lab 1. Antioxidants in Human Health and Disease 2. Mechanisms of Neurodegeneration 3. Artefacts in Cell Culture 4. Ageing in the nematode C.elegans Antioxidants, Nutrition and Ageing Oxidant – Antioxidant Balance AA Metabolism Peroxiredoxins Glutathione system SOD, Catalase Phagocytes Diet-Derived Antioxidants Mitochondrial Blood Respiration components Other electron Iron chelators Albumin, transport chains NADPH oxidases Caeruloplasmin, DUOxes Transferrin, haptoglobin etc Xanthine oxidase Haem proteins So some oxidative damage is inevitable and repair &/or replacement of damaged molecules is essential Antioxidants, Nutrition and Ageing OXIDATIVE DAMAGE AND LOSS OF COGNITIVE FUNCTION • There is strikingly-elevated oxidative damage to all types of biomolecules in the brain in Alzheimer’s Disease (AD) • Oxidative damage is already elevated in mild cognitive impairment, by several biomarkers • Thus it may precede AD development • But why have antioxidant therapies to delay onset or progression of AD not been employed? Halliwell B (2001) Role of free radicals in the neurodegenerative diseases. Therapeutic implications for antioxidant treatment. Drugs & Aging 18, 685 –716. Halliwell B (2006) Oxidative stress and neurodegeneration; where are we now? J. Neurochem. 97, 1634-58. Antioxidants, Nutrition and Ageing A ROLE FOR ANTIOXIDANTS? Risk Factors for AD • ApoE status (plus other genetic factors) • Low intake of folic acid • Hypercholesterolaemia (causes oxidative stress) • Repeated brain trauma (e.g. dementia pugilistica) • Low level of education • Poor language skills / lack of mental exercise • Lack of physical exercise • Diets low in certain polysaturated fatty acids??? • Low dietary intake of antioxidants Antioxidants, Nutrition and Ageing Mechanism of Neurodegeneration CAN IT BE SLOWED BY DIETARY OR SYNTHETIC COMPONENTS? HOW TO STUDY THIS? 1. Cell culture 2. Animal models 3. Humans Antioxidants, Nutrition and Ageing When I was young, the Free Radical –Antioxidant Field was SIMPLE Free radicals are bad Antioxidants are good Taking antioxidants will prevent disease Since free radicals are implicated in ageing, antioxidants will make you live longer Halliwell B (2012) Free radicals and antioxidants: updating a personal view. Nutr Reviews. 70:257-265 Halliwell B (2012) The antioxidant paradox: less paradoxical now? Brit J Clin Pharmacol. In press. Antioxidants, Nutrition and Ageing Protective Plasma Levels 50 M vit. C 30 M vit. E (E:CL ratio > 5.2 mol/mmol) 0.4 M β-carotene Gey (1995) J Nutr Biochem 6, 206-236 Antioxidants, Nutrition and Ageing Protective Plasma Levels BUT BE CAREFUL THESE ARE A MARKER OF A DIET RICH IN FRUITS AND VEGETABLES, WHICH IS ITSELF PROTECTIVE AGAINST DISEASE It could be any component or mixture of components in that diet that is protective Antioxidants, Nutrition and Ageing Plasma levels of E, C and beta- carotene could reflect intake of • Other tocopherols / tocotrienols • Other carotenoids • Flavonoids • Fibre • Phase II inducers • Inhibition / upregulation of cytochromes P450 • Anti-and pro-apoptotic agents • Any other product in the plant Antioxidants, Nutrition and Ageing When I was young, it was SIMPLE Taking Antioxidants will Prevent Disease (with several exceptions) Antioxidants, Nutrition and Ageing BUT SEVERAL TRIALS DID SUCCEED! THE ASAP TRIAL Six year trial of E plus slow release C in 520 hypercholesterolaemic men and women, carotid atherosclerosis progression Significant effect in men (33 % reduction) Nonsignificant effect in women (14 %) Circulation. 2003;107:947-953. Antioxidants, Nutrition and Ageing Int. J. Cancer (2010) 127, 1875-1881 The Supplementation in Vitamins and Mineral Antioxidants Study was a double-blind, placebo-controlled, randomized trial, in which 12,741 French adults (7,713 women aged 35–60 years and 5,028 men aged 45–60 years) received a combination of ascorbic acid (120 mg), vitamin E (30 mg), -carotene (6 mg), selenium (100 lg) and zinc (20 mg), or placebo daily for a median follow-up time of 7.5 years [October 1994 to September 2002]. Antioxidant supplementation decreased total cancer incidence and total mortality in men. Antioxidants, Nutrition and Ageing Limited efficacy in Alzheimers disease No effect in mild cognitive impairment No effect on cardiovascular disease No beneficial effect on cancer Similar failures with vitamin C and -carotene Suggestions of deleterious effects in some studies Antioxidants, Nutrition and Ageing BMJ (2010) 341:c5702 Conclusion In this meta-analysis, vitamin E increased the risk for haemorrhagic stroke by 22% and reduced the risk of ischaemic stroke by 10%. This differential risk pattern is obscured when looking at total stroke. Given the relatively small risk reduction of ischaemic stroke and the generally more severe outcome of haemorrhagic stroke, indiscriminate widespread use of vitamin E should be cautioned against. Antioxidants, Nutrition and Ageing Intervention Trials with Vitamin E • Vitamin E works much better in (at least some) rodent models of neurodegeneration (e.g. APP, ALS) and cardiovascular disease • And it also decreases oxidative damage (measured as F2-isoprostanes) in these models VALIDITY OF RODENT MODELS OF STROKE AND NEURODEGENERATIVE DISEASE? By courtesy of John Milner, National Cancer Institute Antioxidants, Nutrition and Ageing THERE ARE MULTIPLE EXPLANATIONS FOR THIS “FAILURE” OF ANTIOXIDANTS LET’S RE-EXAMINE OUR STARTING HYPOTHESIS • Steady-state levels of oxidative DNA damage contribute significantly to the major cancers. (PROBABLY TRUE) Halliwell B (2007) Oxidative stress and cancer: have we moved forward? Biochem. J. (2007) 401, 1-11. • Steady-state levels of lipid peroxidation contribute significantly to cardiovascular disease (POSSIBLY TRUE) and to neurodegenerative disease (PROBABLY TRUE) Halliwell B (2006) Oxidative stress and neurodegeneration; where are we now? J. Neurochem. 97, 1634-58. Libby et al (2011) Progress and challenges in translating the biology of atherosclerosis. Nature 473, 317-25. Antioxidants, Nutrition and Ageing IN CARRYING OUT THE INTERVENTION TRIAL, DID WE DECREASE OXIDATIVE DAMAGE IN THE SUBJECTS? Because if we did not, no effect would be predicted This was assumed, but not tested Halliwell B (1999) Establishing the significance and optimal intake of dietary antioxidants. The biomarker concept. Nutr. Rev. 57: 104-113. Antioxidants, Nutrition and Ageing Our Studies and Many Others using Biomarkers of Lipid Peroxidation and Oxidative DNA Damage show NO DECREASE on Supplementing Healthy Volunteers with Ascorbate, Beta-Carotene or Vitamin E. BUT effects on “rancid” individuals rarely considered NOR genetic background or racial type Antioxidants, Nutrition and Ageing THERE ARE MULTIPLE EXPLANATIONS FOR THIS “FAILURE” OF ANTIOXIDANTS • They ignore individual differences in “rancidity” (which depend on age to some extent) Increased isoprostanes in thalassaemia Antioxidants, Nutrition and Ageing RESPONSES MAY ALSO CHANGE WITH RACIAL ORIGIN. Vorinostat Histone deacetylase inhibitor under evaluation as anti-cancer drug Vorinostat Glucuronide Glucuronidation (UGT2B17) (less activity) Vorinostat Hydrolysis followed 4-Anilino-4-oxobutanoic acid by β-oxidation UGT2B17*2 (del variant) •Reduced UGT2B17 activity •Homozygotes: 10% Caucasians Wong et al. Pharmacogenetics & Genomics 60-70% Asians 2011, 21(11):760-8 UGT2B17*2 homozygotes present in 62% of our cohort RESPONSE TO NUTRIENTS IS ALSO LIKELY TO BE DIFFERENT Slide courtesy of Prof John Wong Antioxidants, Nutrition and Ageing THEY MAY ALSO VARY WITH GENETIC BACKGROUND AND AGE The gene encoding HAPTOGLOBIN (a haemoglobin binding protein) is polymorphic with 3 genotypes, 2-2, 2-1, 1-1. The 2 allele is a poorer antioxidant against haemoglobin-induced oxidative damage in vitro. Arterioscler Thromb Vasc Biol. 2008; 28:341-347 Atherosclerosis 2011; 219:240-244 Pharmacogenomics 2010; 11:675-684 Antioxidants, Nutrition and Ageing THE FLAVONOID STORY • Polyphenols have powerful antioxidant effects in vivo. • Polyphenols are not-fully absorbed and are rapidly metabolised, blocking the antioxidant –OH groups. • Phenolic metabolites usually have lower antioxidant activity • Levels of phenolics are low in the face of all the other plasma antioxidants. • Their bioavailabilty to the brain is poor but consumption of flavonoid-rich foods seems to enhance cognition in some studies It is unlikely that Flavonoids act as Systemic Antioxidants In Vivo Halliwell B. (2008) Are polyphenols antioxidants or pro-oxidants? What do we learn from cell culture and in vivo studies? Arch Biochem Biophys. 476, 107-112 Antioxidants, Nutrition and Ageing FLAVONOIDS AND OTHER ANTIOXIDANTS MAY ACT DIRECTLY IN THE GASTROINTESTINAL TRACT Halliwell et al (2000) Free Rad Res 33, 819-830 The GI tract: a major site of antioxidant Action? Jenner AM, Rafter J, Halliwell B (2005) Free Radic Biol Med. 38, 763-772 Human fecal water content of phenolics: The extent of colonic exposure to aromatic compounds. Antioxidants, Nutrition and Ageing Antioxidants Work in the GI Tract! STOMACH SMALL COLON/RECTUM INTESTINE Duodenum Ileum Jejenum High concentration of ascorbate in food Vitamin C completely absorbed* Little vitamin C present* and
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