1 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /1, No. -, 3-33 (,*+*)

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1 1 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /1, No. -, 3-33 (,+) 93 E# ect of Awamori Lees on Growth, Serum and Liver Lipid Concentration in Rats Fed a High Cholesterol Diet Toshitsugu Nohara, Shuntoku Uechi, Goh Ogura, Kunioki Hayashi and Katsunori Sunagawa United Graduate School of Agriculture Sciences, Kagoshima University, Korimoto +,+,., Kagoshima 23 0/ Department of Subtropical Biochemistry and Biotechnology, Faculty of Agriculture, University of the Ryukyus, Senbaru +, Nishihara, Okinawa 3-,+- Department of Subtropical Agro-Environmental Sciences, Faculty of Agriculture, University of the Ryukyus, Senbaru +, Nishihara, Okinawa 3-,+- Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, +,+,. Korimoto, Kagoshima 23 0/ Awamori lees (AL), a by-product generated during manufacturing of Awamori, is rich in protein, dietary fiber and organic acids. As large amounts of AL have been dumped to date, this study attempted to develop a usage for AL as a feedstu# and/or as a functional food material. We investigated the e# ect of AL on growth and serum and liver lipid concentrations in /-week-old rats fed a diet with - cholesterol for, weeks followed by diets supplemented with /,,/ and / AL for. additional weeks. The body weight gain and feed intake for each AL-supplemented diet group was the same as that for the control group. Liver weight, and serum and liver total cholesterol concentrations were dose-dependent decreased in the ALsupplemented groups ; however, the serum HDL-cholesterol level was una# ected by AL intake. The / AL group showed significantly lower serum and liver triglyceride concentrations. Fecal weight increased with the amount of AL in the diet, with a /-fold increase in total bile acid content in the / AL group. These results show that AL can e# ectively reduce lipid concentrations, suggesting its use as feedstu# and/or functional food material. (Received Aug. -+,,3 ; Accepted Nov. +-,,3) Keywords : Awamori lees, fatty liver rat, dietary fiber, cholesterol, bile acid :, /2 +,+,. 3-, , /2 +,+,. Corresponding author uechis@agr.u-ryukyu.ac.jp

2 94 /1 -,+ - 2., Prosky., Table + /,/ / ww - -. SD, -, // / +, a-cornstarch Casein Sucrose Soy bean oil Cellulose powder Mineral mix AIN-3- G Vitamin mix AIN-3- L-Cystine Choline chloride Cholic acid sodium salt Cholesterol Awamori lees , 4,/ / , 4,/ -4 / , 4,/ , 4,/, , 4,/ /4 / AIN-3- CE-kg - +/ + / + Standard,.,. Control /,/ / / AL,,/ AL / AL. - rpm, DW-. TC 1,. FC HDL HDL,- Table + Composition of experimental diets +, (w/w of diet) Standard Control / AL,/ AL / AL +,,, Control : standard with - cholesterol. Experimental diets : standard with / to / Awamori lees (AL).

3 3 : 95 TG PL E E HDL E 2 E C SAS Duncan / 0 Folch TL TC TG / + + X-+ Standard Control /,/ / Fig. + 0 Control /,/ / AL -,. Standard. - Table, Control Standard 1 Eaton /,/ / 0/ + - AL - -ao -ao - -. nm NADH 1,, /,. M ,./ mm Tris-HCl, ph / rpm + +/, ma + V Fig. + E# ect of the Awamori lees (AL) on the growth in rats fed a high cholesterol diet SDS-PAGE Molecular Each value represents the mean of / rats., Standard ; Weight Standards, Broad Range, Control ;, / AL ;,,/ AL ;, / AL. Table, E# ects of Awamori lees (AL) on body weight, feed intake and liver weight in rats fed a high cholesterol diet Standard Control / AL,/ AL / AL Initial body weight (g), ,/34, +/43,/341 +/41,/242 +,4, Final body weight (g) -324+,, , ,-4, -140 Body weight gain (g/. weeks) +/ /, ,24+ +0,42,24+ Feed intake (g/day),.43.4,, ,/4--41,14.4+,.42,43 Total feed intake (g/. weeks) 0314, , ,4. 1/ / 243 Liver weight (g) +.4,0 +41/ +341.,4/. +34,1, , b a a ab b Values are meanss.d. of / rats. Means in the same column with di# erent superscripts are significantly di# erent ( p4/ ).

4 96 /1 -,+ - 4 Table - E# ects of the Awamori lees (AL) on serum lipid concentrations in rats fed a high cholesterol diet Concentration (mg/dl) TC HDL FC TG PL Standard Control / +, , 124, -40 +, bc bc ab bc / , ,4/ +,34/,14, a a a ab / AL 1.4/ / ,43 +, ,,.4, b b a a,/ AL /04, / ++,43,-43 c bc ab abc / AL / , , , c c b c Values are meanss.d. of / rats. Means in the same column with di# erent superscripts are significantly di# erent ( p4/ ). Abbreviations, TC : Total cholesterol, HDL : HDLcholesterol, FC : Free cholesterol, TG : Triglyceride, PL : Phospholipid. Table. E# ects of the Awamori lees (AL) on liver lipid Table / E# ects of the Awamori lees (AL) on fecal weight, concentrations in rats fed a high cholesterol diet moisture and total bile acid content in rats fed Concentration (mg/g wet liver weight) a high cholesterol diet TL TC TG Dry fecal Fecal Fecal total weight moisture bile acid Standard cd +,/ c.4.,4. b +,4+.4- (g/- days) ( ) (mmol/l of - days) Control, /, ,4+ a a a / AL,,+4.,41 +/ / 14- ab b a,/ AL / bc b a / AL ++, ,4.,,4 /4/ d c b Standard Control ,+ 4+2 d b b / / 14, 4,3 4,. cd b b / AL 14, 43, ,0 c b b,/ AL +, ,2 b a a Values are meanss.d. of / rats. Means in the same / AL /0 4/2 a a a column with di# erent superscripts are significantly Values are meanss.d. of / rats. Means in the same di# erent ( p4/ ) 4 Abbreviations, TL : Total lipid, TC : column with di# erent superscripts are significantly Total cholesterol, TG : Triglyceride. di# erent ( p4/ ).,/ AL / AL - / AL Control / Standard Fig.,, + +. kda Table - TC FC /,/ / AL -, -, kda,-kda HDL -+ kda TG PL / AL - Table. TL,/ AL / AL , TC / TG / AL. - - Table / -

5 5 : 97 Fig., SDS-PAGE of Awamori lees protein Lane + : Awamori lees protein, lane, : Thai rice protein, M : molecular size marker / Control TL,2.- / 2 -,/ / AL Standard TL +3./ ++./ TG TC HDL Standard TG Control / AL Control, 22+ kcal / AL,/ AL / /,/ /. AL, /3, 10,,./3 kcal TC TG Control - / AL +, TC TG Tsutsui +1 in vitro 2 AIN-3-G tunicase pepsin-pancreatin / Tsutsui Wistar / Tanaka, Morita +3, + RPI,+ Robbins TC TG

6 98 /1 -,+ - 6 / AL Control -.. / -, / AL Control /.. + g /,/ /. / AL.-0 mm Control.. mm / Control 2,2.- ph,/ / AL Standard +. kda TC I +. kda -+ /,/ / AL HDL Standard kda +. kda TG Control / AL Control / Tanaka +2 I,, Lin,, p. 10,2, // p. +2, / 01,/. Prosky, L., Asp, N.G., Schweizer, T.F., DeVries, J.W. and Furda, I., Determination of insoluble, and total dietary fiber in foods and food products : interlaboratory study. / J. Assoc. O #. Anal. Chem., 1+, ++1 +,- ( +322). / Philip, G.R., Forrest, H.N. and Geroge, C.F., AIN-3purified diets for laboratory rodents : Final report of the American institute of nutrition ad hoc writing committee on the reformulation of the AIN-10A rodent diet. TC TG J. Nutr., +,-, /+ ( +33-). 0 Folch, J., Lees, M. and Sloane-Stanley, G.H., A simple method for the isolation and purification of total lipids from animal tissues. J. Biol. Chem.,,,0,.31 /3 ( +3/1). 1 Eaton, D.L. and Curtis, D.K., E# ects of acute administration of taurocholic and taurochenodeoxycholic acid on biliary lipid excretion in the rat. Proc. Soc. Exp. Biol. Med., +/+, +32,, ( +310). 2

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