Iodine-enriched egg reduced total body fat and visceral fat among normal individuals: A placebo-controlled, randomized, double-blind study

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1 Oriinal Article Glycative Stress Research Online edition : ISSN Print edition : ISSN Received : Auust Accepted : September 1, 2016 Published online : September 30, 2016 Iodineenriched e reduced total body fat and visceral fat amon normal individuals: A placebocontrolled, randomized, doubleblind study Jiro Yokoyama 1), Meumi Morioka 1), Hajime Inoue 2), Yoshikazu Yonei 3), Keiichi Suzuki 4), Kazuo Katoh 4) 4, 5), SanGun Roh 1) Nippon Nosan Corporation, Yokohama, Kanaawa , Japan 2) Dept. of Plastic & Reconstructive Surery, Div. of Stem Cell Medicine (Endowed Chair by ANGFA Co.) St. Marianna University School of Medicine, Kawasaki , Japan 3) AntiAin Medical Research Center / Glycative Stress Research Center, Graduate School of Life and Medical Sciences, Doshisah University, Kyoto, Japan 4) Division of Functional and Developmental Science of Livestock, Graduate School of Aricultural Science, Tohoku University, Aoba, Sendai , Japan 5) Lab of Animal Physioloy, Graduate School of Aricultural Science, Tohoku University, Sendai , Japan Abstract Objectives: This study aimed to evaluate the effects of iodineenriched e () on total body fat, visceral fat, body weiht, body mass index (BMI), and safety parameters amon normal individuals. Methods: This randomized, doubleblind, placebocontrolled study recruited 56 normal individuals ( men, women; BMI: k/m 2 ), who were assined to receive either boiled or ordinary e () each day for 12 weeks. Total body fat and visceral fat were measured at baseline and 12 weeks usin abdominal computed tomoraphy. Body weiht, BMI, and blood samples were also evaluated at baseline and week 12. Results: Althouh both roups exhibited similar caloric intakes, the roup exhibited sinificantly reduced visceral fat area at week 12, compared to the roup. In addition, the roup exhibited a sinificant reduction in total and visceral fat area, compared to the baseline value. There were no sinificant differences in the blood parameters between the and roups. No sinificant adverse effects were observed. Conclusion: These results suest that inestion may help reduce visceral fat. KEY WORDS: iodineenriched e, total body fat, visceral fat Introduction Body fat accumulation is reulated by enery intake and expenditure, and a serious imbalance between these two processes is a major risk factor for cardiovascular disease and lifestylerelated diseases, such as diabetes, hypertension, hyperlipidemia, and arteriosclerosis. In this context, metabolic syndrome is defined as a roup of disorders that involve abdominal obesity with hih blood lucose levels, dyslipidemia, and hih blood pressure. Several functional foods and substrates can be used to prevent the accumulation of excess total body fat and visceral fat, and these functional foods have been commonly incorporated into diets 1). Functional es are easily consumed every day, and previous studies have revealed that iodineenriched es () protect aainst diabetes and alter lipid metabolism 26). Furthermore, patients with diabetes who consumed for 3 months exhibited sinificantly lower blood lucose levels, compared to normal individuals 7). Moreover, chickens that received e white from exhibited lower plasma lucose concentrations, compared to chickens that were fed e white from ordinary es () 8). However, to the best of our knowlede, no studies have evaluated the effects of and on body fat accumulation in humans. Therefore, the present study aimed to evaluate whether inestin helped reduce total body fat and visceral fat in normal individuals. Correspondin author: Sanun Roh, Lab of Animal Physioloy, Graduate School of Aricultural Science, Tohoku University, Sendai , Japan Tel: Fax: sanun. roh@tohoku.ac. jp Coauthors: Yokoyama J, j_yokoyama@nosan.co.jp ; Morioka M, m_morioka@nosan.co.jp ; Inoue H, h2inoue@mariannau.ac.jp ; Yonei Y, yyonei@mail.doshisha.ac.jp ; Suzuki K, k1suzuki@bios.tohoku.ac.jp ; Katoh K, kato@bios.tohoku.ac.jp _ 172 _ Glycative Stress Research 2016; 3 (3): (c) Society for Glycative Stress Research

2 Glycative Stress Research Methods Ethical considerations The present study complied with the ethical principles of the Declaration of Helsinki (the 2004 clarification by the World Medical Association General Assembly), Japan s Act on the Protection of Personal Information, the Ministry of Health, Labor, and Welfare s Ordinance reardin Good Clinical Practice (no. ; March 27, 1997), and the Ethical Guidelines for Epidemioloical Research (Ministry of Health, Labor, and Welfare; Ministry of Education, Culture, Sports, Science, and Technoloy). The study s desin was approved by the Doshisha University Ethics Committee for Scientific Research Involvin Human Subjects (##14082) and the clinical research ethics board of the Tokyo Synery Clinic. All participants provided their informed consent after receivin a full explanation reardin the study. Study desin This study used a randomized, doubleblind, placebocontrolled desin. Participants were recruited from individuals who were identified by the Department of AntiAin Medicine (Doshisha University, Faculty of Life and Medical Sciences). Individuals were considered eliible if they were 20 years old and did not fulfill any of the exclusion criteria: a food or dru allery, prenant or lactatin, receivin treatment usin medication or followup, diabetes mellitus dianosis, pronounced cardiopulmonary function disorder, receivin medication for hypertension, a history of astrointestinal surery, or suspicion of infectious disease. Some individuals were also excluded from the study based on the investiator s discretion. The included individuals were randomly allocated to receive one boiled (Yodoran Hikari ) or one each day for 12 weeks. The compositions of and were shown in Table 1. Durin the study, the participants were required to make five clinic visits: a health screenin, a baseline evaluation, after 4 weeks of inestion, after 8 weeks of inestion, and at week 12 (4 weeks after the inestion period). A welltrained nurse recorded the participants body weiht, heiht, BMI, and subjective complaints at each visit, and collected blood samples for laboratory testin (collected durin the mornin after a 10 h fast). The blood test parameters included levels of trilycerides, total cholesterol, hihdensity lipoprotein cholesterol, lowdensity lipoprotein cholesterol, and other metabolites. Selfreported dietary caloric intakes for the 3 consecutive days before the baseline and week 12 visits were assessed, and the averae daily caloric intake values were calculated for each participant. Subcutaneous and visceral fat assessment Computed tomoraphy (CT) was performed at baseline and week 12 to measure the subcutaneous and visceral fat areas at the level of the umbilicus (L4 L5; Leato Duo CT; GE Yokoawa Medical Systems, Tokyo, Japan). The visceral fat area was used to evaluate visceral obesity. The CT data were analyzed usin FatScan software. Statistical analyses All data were reported as mean standard error. The paired ttest was used to evaluate intra and interroup differences. A Pvalue of < 0.05 was used to identify statistically sinificant differences. Results and Discussion Amon the 64 individuals who were randomized to the treatment roups, 5 individuals dropped out because of personal reasons that were unrelated to the or. Fiftynine individuals completed the study, althouh 3 individuals were excluded durin the analysis because of poor compliance (1 individual did not follow the inestion protocol) or unreliable CT data (n = 2). Thus, the final analyses evaluate data from 56 individuals ( men, women, ae: years, body mass index [BMI]: k/m 2 ). The and roups exhibited similar baseline demoraphic and anthropometric characteristics (Table 2). The FatScan analyses of the CT data revealed that the roup exhibited sinificantly lower week12 values for total body fat and visceral fat, compared to the roup (p = and p = 0.001, respectively). However, both roups exhibited similar week12 values for BMI, trilyceride levels, lowdensity lipoprotein cholesterol levels, and hihdensity lipoprotein cholesterol levels (Table 3). Most blood test parameters did not exhibit timerelated intraroup chanes (e.., the roup exhibited similar baseline and week12 values for free triiodothyronine and thyroxine). However, the roup did exhibit a sinificant 12week increase in urinary iodine levels. Our previous study revealed that the treatment of 3T3L adipocytes usin yolk from reduced their fat accumulation and inhibited adipocyte differentiation 9). Another study also revealed altered lipid metabolism in rats after they inested, such as hiher tissue lipoprotein lipase activity 3). The present study showed that the pituitarythyroid axis was not chaned by inestion of. The iodide related components as iodide peptide of responsible for fat accumulation in adipocytes are present in e yolk. These findins suest that the iodide peptide in may help directly reduce fat accumulation in normal individuals, without chanin their thyroid hormone secretions. Conclusion The present study revealed three main findins. First, the roup exhibited sinificantly lower total body fat and visceral fat, compared to the roup. Second, both roups exhibited similar values for body weiht, BMI, and blood parameters (e.., total cholesterol and lowdensity lipoprotein cholesterol). Third, none of the participants reported problematic adverse effects. Conflicts of interest The authors have no financial relationship with the oranization that sponsored this study. The sponsor played no part in the research, data collection, data analysis, writin of the manuscript, or decision to publish. _ 173 _

3 Table 1. The composition of and I E Crude protein Crude fat Crude Carbohydrate Enery kcal Iodine μ Iodide peptide μ 361 ND, iodineenriched e;, ordinary e; ND, Not detected All compositions were shown per 100 rams of edible e. Table 2. The participants' baseline characteristics (n = ) (n = ) Total (n = 56) Ae years Heiht cm Body weiht k Body mass index k/m Total fat area Subcutaneous fat area Visceral fat area White blood cells /μl 5, , , , , , Red blood cells 10 4 /μl Hemalobin /dl Hematocrit % Trilycerides LDL HDL Free T 4 n/dl Free T 3 p/ml μ/l corrected for creatinine μ/ Cre Total free iodine μ/day , Data are expressed as mean standard error., iodineenriched e;, ordinary e; HDL, hihdensity lipoprotein cholesterol; LDL, lowdensity lipoprotein cholesterol; T3, triiodothyronine; T4, thyroxine. _ 174 _

4 Glycative Stress Research Table 3. The characteristics of participants who inested iodineenriched e and ordinary e for 12 weeks Ae Heiht Body weiht Body mass index Total fat area Subcutaneous fat area Visceral fat area White blood cells Red blood cells Hemalobin Hematocrit Total cholesterol Trilycerides LDL HDL Free T4 Free T3 corrected for creatinine Total urinary idoine years cm k k/m 2 /μl 10 4 /μl /dl % n/dl p/ml μ/l μ/ Cre μ/day , , Baseline week , , , Data are expressed as mean standard error. 1 Pvalues for comparin baseline and week 12 values in the and roups. 2 Pvalues for comparin the 12week overall chane between the and roups. 3 Pvalues for comparin the 12week percent chane between the and roups., iodineenriched e;, ordinary e; HDL, hihdensity lipoprotein cholesterol; LDL, lowdensity lipoprotein cholesterol; T3, triiodothyronine; T4, thyroxine , , , , , , Pvalues _ 175 _

5 References 1) So KH, Suzuki Y, Yonekura S, et al. Soluble extract of soybean fermented with Asperillus oryzae GB107 inhibits fat accumulation in cultured 3T3L1 adipocytes. Nutr Res Pract 2015; 9: ) Katamine S, Hoshino N, Totsuka K, et al. Effects of the lonterm (79 months) feedin of iodineenriched es on lipid metabolism of rats. J Nutr Sci Vitaminol (Tokyo) 1983; 29: ) Katamine S, Hoshino N, Totsuka K, et al. Effects of the lonterm (1719 months) feedin of hihiodine es on lipid metabolism and thyroid function in rats. J Nutr Sci Vitaminol (Tokyo) 1985; 31: ) Tanaami S, Katamine S, Hoshino N, et al. Histopatholoical study on rats fed iodineenriched es lonterm (7 and 19 months). J Nutr Sci Vitaminol (Tokyo) 1985; 31: ) Katamine S, Hoshino N, Totsuka K, et al. Effect of Iodine Enriched Es on Lipid Metabolism of Growin Rats at Exercise and Meal. Nutrition and Food (Eiyo Syokuryou) 1981; 34: ) Iwama M, Iitoi Y, Takahashi N, et al. Effect of Iodine Enriched Es on Metabolic Activation of Benzo[a] pyrene to Mutaens and DruMetabolizin Enzyme Activities in Rat Hepatic Tissue. Food Hyiene and Safety Science (Shokuhin Eiseiaku Zasshi) 1989; 30: (in Japapnese). 7) Yokoyama J, Shimokawa M, Tomioka M, et al. Influence of insulin index by lon term treatment of iodine enriched e in Japanese. New Food Industry 2010; 52: 2126 (in Japanese). 8) Ito K, Yokoyama J & Kita K. Suppression of intestinal lucose absorption by administratin iodineenriched es to chickens as hyperlycemic experimental animals. Antiain medicine 2014; 10: ) Yokoyama J, So KH, Suzuki Y, et al. Antiobese effect of iodineenriched yolk in cultured adipocytes. Nihon Chikusan Gakkaiho 2016; (In press) (in Japanese). _ 176 _