Sunghyen Lee 1, Youngmin Lee 2, Sungjoon Lee 3, Haejeung Lee 4, Hyun Lillehoj 5

Transcription

1 Dec 8, 2015 Sunghyen Lee 1, Youngmin Lee 2, Sungjoon Lee 3, Haejeung Lee 4, Hyun Lillehoj 5 1 Functional Food & Nutrition Division, National Institute of Agricultural Science, 2 Seoul Women s Univ, 3 Eulji Univ, 4 Korea Univ, Republic of Korea, ARS-USDA, USA 1

2 Contents Introduction 2/39 Objective - 12 Materials & Methods, Results * black rice extract (BRE), post-menopausal model in vitro 13/39 in vivo - 20 human study - 30 Summary 35/39 Discussion - 36 Conclusion 37/39 2

3 Introduction Rice (Oryza sativa L.) is widely consumed throughout the world and rice consumers have been paying attention to various values of rice. With growing concerns regarding health, black, purple, and red rice with unique flavor, nutritional and biological properties have been developed. 3

4 Introduction The supplementation of black rice and its anthocyanin pigment reduced atherosclerotic lesions in hypercholesterol-emic animal models. Dietary black rice supplementation alleviated hepatic steatosis by increasing fatty acid oxidation in the mice fed a high fat diet. Black rice is a kind of brown rice and is not well digested. So most of functional materials existing in aleurone layer of black rice are hard to be observed. Bio-functional effects of black rice have been tested with it s extracts and major compounds were known as polyphenols and flavonoids. 4

5 Introduction * Thus numerous in vitro and in vivo studies have shown us that black rice may be preventive against chronic diseases by reducing fat weight or its ratio in the body. 5

6 Introduction The average life span has been increasing. 6

7 Introduction Obesity % is higher in women than in men for over 60 years old peoples. Comparison of obesity percentage by age and gender 7

8 Introduction For post-menopausal women, body fat tends to shift to the abdomen and body composition changes with higher ratio of fat in the body. The women experience increased risk of metabolic syndrome and heart disease. 8

9 Introduction For post menopausal women, keeping good health is so important because their health condition continues to/decides the health condition of their old age. Exogenous estrogen has been shown to be protective against metabolic abnormalities. But longterm usage of hormone replacement therapy may increase the risk of breast cancer and cardiovascular diseases in postmenopausal women. 9

10 Introduction People have been interested in natural foods such as black rice to prevent and control obesity or body fat accumulation. Hypothesis 1. Extracts from aleurone layer of black rice (BRA) may effectively prevent obesity and reduce body fat accumulation in the menopausal women. 10

11 2. BRA is 8% of black rice. If we use BRA rather than whole black rice as brown rice for functional material extraction, we can save ethanol as extraction solvent and reduce its amount to 1/13. Oryza sativa L. Brown rice 8.0% aleurone layer (BRA) 92.0% milling rice yields Black rice extract (BRE) 11

12 Objective This study was conducted to evaluate suppressive effects of black rice aleurone layer extract (BRE) on body fat, serum lipid, and hormone levels in ovariectomized model. 12

13 Materials and Methods in vitro study 1 2 Preparation of extracts Oryza sativa L. Brown rice (BR) 3 8.0% aleurone layer (AL) 92.0% milling rice yields (white rice, WR) + 50, 70%%(v/v) EtOH 40 o C, 48 h

14 fold (X) Results - in vitro 개선 Black 전 rice 기술 ( 흑미현미 ) 개선 Black 후 rice 기술 aleurone ( 흑미호분층 : A layer + B + C + D) Cyanidin 3- glucoside Peonidin 3- glucoside 전체 Total anthocyanin Fig. Comparison of anthocyanin contents in black rice and Black rice aleurone layer 14

15 Experimental design - in vitro Comparing - part of rice: BR, AL, WR - ethanol concentration : 50%, 70% Cells: 3T3L1 Factors : cell differentiation, gene expression, lipid concentration 15

16 Results - in vitro Control BR AL Control BR AL Fig. Comparison of BR and AL effect on adipocyte differentiation in 3T3L1 cells 16 Fig. Effects of BRE-AL on adipocyte differentiation in 3T3L1 cells

17 Results - in vitro Fig. Effects of BRE-AL on adipocyte differentiationrelated gene expression in 3T3L1 cells 17

18 Results - in vitro Cell differentiation - Oil Red O Fat oxidation and synthesis Fig. Effects of BRE-AL on adipocyte differentiation, fatty acid oxidation, and its synthesis in 3T3L1 cells 18

19 Results - in vitro Fig. Effects of BRE-AL on lipid accumulation-related gene expression in 3T3L1 cells at 9 th day 19 19

20 Chosen material for in vivo and human studies 1 2 Preparation of BRE Oryza sativa L % aleurone layer (BRE, BRE-AL) 92.0% milling rice yields + 50%(v/v) EtOH 40 o C, 48 h Brown rice Black rice extract (BRE) 20

21 Experimental design - in vivo Metabolic changes induced by estrogen depletion from ovariectomy share many similar characteristics with changes such as weight gain, increased adiposity, and the development of fatty liver in menopausal women. -3 week 0 week 12 week (Adaptation period) 5-week-old SD rats Operation Sham or Ovariectomy Diet: High fat diet (at calories 45%) Groups Sham OVX Control I.F 10mg/kg B.W BRE 30 or 90mg/kg B.W 21

22 Measuring factors - in vivo Body weight gain, fat pad weight Adipocyte size (adipocyte index) Serum triglyceride, total-, LDL-, HDL-cholesterol Serum leptin, adiponectin concentration Hepatic and fecal total lipids, triglyceride, total cholesterol Statistical analysis Windows SAS package program (ver. 9.2) One-way ANOVA ( =0.05) Duncan s multiple range test 22

23 Body weight gain (g/12 weeks) Results - in vivo a ab b b 150 c Sham OVX IF BRE-30 BRE-90 Fig. Treatment effects of BRE-AL on body weight gain of ovariectomized rats 23

24 Weight Results - in vivo Sham OVX IF BRE-30 BRE-90 a b b b c a b bc bc 10 5 ab a ab b b c 0 Perirenal fat Abdominal fat Total body fat Fig. Treatment effects of BRE-AL on body fat weight of ovariectomized rats 24

25 *Adiposity Index (%) Results- in vivo b a b b b Sham OVX IF BRE-30 BRE-90 Fig. Treatment effects of BRE-AL on adiposity index of ovariectomized rats, * fat weight/body weight 25

26 Concentration (mg/dl) Results- in vivo Sham OVX IF 160 BRE-30 BRE-90 NS ab a bc a c Triglyceride Total cholesterol Fig. Treatment effects of BRE-AL on serum lipid profiles of ovariectomized rats 26

27 Concentration (μg/ml) Results- in vivo b ab a a a Sham OVX IF BRE-30 BRE-90 b b c bc bc 2 0 Adiponectin Leptin Fig. Treatment effects of BRE-AL on serum adiponectin and leptin levels of ovariectomized rats 27

28 Concentration (mg/g) Results- in vivo a a a a Sham OVX IF BRE-30 BRE-90 b a a a a b NS Total lipids Triglyceride Total cholesterol Fig. Treatment effects of BRE-AL on hepatic lipid profiles of ovariectomized rats 28

29 Experimental design - human study Body weight and fat content Measure Food and nutrient intake factor BRE-AL Placebo Number Age (46 ~69) Body weight

30 Experimental design - human study Treat with BRE-AL: mg/capsule, 2 capsules/day, 12 weeks Factors: fat weight, - ratio -Bioelectrical impedance analysis (BIA), -Dual-energy x-ray absorptiometry (DEXA), -Computed tomography(ct) Statistical analysis: SASV(version 9.4) 30

31 Body fat ratio (%) Results Human study BIA Fat ratio BIA Body fat ratio P=0.044 P= wk 12wk 흑미 BRE-AL 플라시보 Placebo Fig. Treatment effects of BRE-AL on fat ratio of post menopausal women

32 Results Human study Abdominal fat weight (kg) DEXA Body fat weight P= P=0.000 P= wk 12wk 흑미 BRE-AL 플라시보 Placebo Fig. Treatment effects of BRE-AL on abdominal fat weight of post menopausal women

33 Results Human study Total fat weight (kg) DEXA- total fat weight P=0.002 P= wk 12wk P= 흑미 BRE-AL 플라시보 Placebo Fig. Treatment effects of BRE-AL on total fat weight of post menopausal women

34 Visceral fat area (cm2) CT visceral fat 10,450 10,250 10,182 P= ,050 9,850 0wk 12wk 9,650 9,450 9,250 9,629 9,481 9,293 9,050 8,850 8,650 흑미 BRE-AL 플라시보 Placebo Fig. Treatment effects of BRE-AL on visceral fat area of post menopausal women

35 Summary < in vitro> BRE-AL treatment significantly decrease adipocyte differentiation and lipid accumulation. < in vivo> Body weight gain, body fat weight, and adiposity index increased in the OVX group, but they significantly decreased in the groups supplemented with IF or BRE-AL. Serum triacylglyceride and leptin levels decreased in BRE-AL groups while serum adiponectin level significantly increased compared to that of the OVX group. The reduced fat weight may be explained by the changed gene expression or hormone levels. < human study> Body fat in the BRE treated group was significantly lower than placebo group. 35

36 Discussion BRE-AL effectively reduced body fat weight in post menopausal model at 30 or 90 mg/kg BW though whole black rice extract showed possibility of fat weight reducing at 200 mg/kg BW (Jang et al., 2015). BRE-AL can be used in people who have been interested in natural foods to control obesity or body fat accumulation. Using BRE-AL can reduce extracting solvent volume, prevent environment, and make white rice as foods. The team has been working to register the extract(bre-al) as a healthy food. 36

37 Conclusion The results suggest that black rice aleurone layer may be a useful food source to decrease obesity and its related diseases by modulating lipid metabolism in estrogen-deficiency model. 37

38 References Yang DS, Lee KS, Jeong OY, Kim KJ, Kays SJ. Characterization of volatile aroma compounds in cooked black rice. J Agric Food Chem. 2008;56: Chiang AN, Wu HL, Yeh HI, Chu CS, Lin HC, Lee WC. Antioxidant effects of black rice extract through the induction of superoxide dismutase and catalase activities. Lipids 2006;41: Min SW, Ryu SN, Kim DH. Anti-inflammatory effects of black rice, cyanidin-3-o-beta-d-glycoside, and its metabolites, cyanidin and protocatechuic acid. Int Immunopharmacol 2010;10: Kim HW, LEE AY, Yeo SK, Chung H, Lee JH, Hoang MH, Kim YS. Metabolic profiling and biological mechanisms of body fat reduction in mice fed the ethanolic extract of black-colored rice. Food Res Int 2013;53(1) Seo SJ, Choi YM, Lee SM, Kong SH, Lee JS. Antioxidant Activities and Antioxidant Compounds of Some Specialty Rices. J Korean Soc Food SciNutr 2008;37(2): Jang HH, Park MY, Kim HW, Lee YM, Hwang KA, Park JH, Park DS, Kwon O. Black rice (Oryza sativa L.) extract attenuates hepatic steatosis in C57BL/6J mice fed a high-fat diet via fatty acid oxidation. Nutrition & Metabolism 2012;9:27-38 Davis SR, Castelo-Branco C, Chedraui P, Lumsden MA, Nappi RE, Shah D, Villaseca P. Understanding weight gain at menopause. Climacteric. 2012;15(5): Kaaja RJ. Metabolic syndrome and the menopause. Menopause Int. 2008;14(1):21-5 Rossouw JE, Anderson GL, Prentice RL et al., Risks and benefits of estrogen plus progestin in healthy postmenopa usal women: principal results From the Women's Health Initiative randomized controlled trial. JAMA. 2002;288( 3):

39 Thank you for your attention! 39