7 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /2, No. +, 1 +/ (,++) 7 * E# ects of Pigment Extracts from Purple Carrot ( Daucus carota, L.) on the Re

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1 7 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /2, No. +, 1 +/ (,*++) 7 * * E# ects of Pigment Extracts from Purple Carrot ( Daucus carota, L.) on the Reactions in Mice Subjected to Restraint Stress Mitsuru Watanabe, Keiichi Musumi* and Jun Ayugase National Agricultural Research Center for Tohoku Region,. Akahira, Shimokuriyagawa, Morioka, Iwate *,* *+32 * Takii Plant Breeding Station, +-0* Hari, Konan, Shiga /,* -,-+ We investigated the e# ects of carotenoid and/or anthocyanin pigment extracts from purple carrot on the reactions induced by restraint stress in mice. Purple carrot contained b-, a-carotene, and phytoene as carotenoid compounds, and five anthocyanin compounds that contained cyanidin as aglycon. Aliquots of carotenoid and/or anthocyanin pigment extracts from purple carrot in an acidic detergent solution (citric acid, Tween 2* ) were administered intragastrically once per day for three days to mice, after which they were restrained for,. h. Unrestrained mice were administered acidic Tween solution with or without free access to feed and water. Restraint stress induced elevation in plasma arteriosclerotic index, hepatic total cholesterol and the amount of thiobarbituric acid-reactive substances (TBARS) of plasma and liver tissues in mice. In contrast, these variables were suppressed in the restraint mice that were administered purple carrot pigment extracts. Furthermore, additional e# ects of carotenoid and anthocyanin pigment extracts on a reduction in hepatic cholesterol level were observed. These results suggest that purple carrot pigment extracts have in vivo anti-stress e# ects against the reactions induced by immobilization in mice. (Received Jun.,+,,*+* ; Accepted Jul.,3,,*+*) Keywords : purple carrot, carotenoid, anthocyanin, restraint stress, anti-stress : Seyle +,. / sympatheti-adrenomedul- lary, SAM system hypothalamic-pituitary-adrenocortical, HPA system 0 *,* *+32. * /,* -,-+ +-0* Corresponding author wata@a# rc.go.jp +/. +0,

2 8 /2 +,*++ + 8,* + + +* ++, *,. g / ml /.,. +, +. TBARS Sep-pak C+2 Sep-pak C Plus Short / ml *+. TFA / ml *+. TFA / ml */. TFA :, : 2 / ml *+. TFA. HPLC LC-++** ESI./** V LC-MS QSTAR Pulsar-i : ODS Symmetry / C +2,,. + mm+/* mm : -/ : 3./,- -* *. + TFA : *., mlmin : /,* nm,2* nm,** nm0** nm - / g :, : + +** ml, +** ml / +** ml, /* ml ++* 0* wv +0 : + : + /, / 2* HPLC APCI +./* LC-MS TCH- 1,+,**2 2 : ODS Symmetry / C+2 +,.. 0 mm,/* mm : -/ : : +/

3 9 : 9, : 2 : +.* mlmin :./* nm 2 +2,2* nm,** nm0** nm Folch. *.. g +0. ml 0 ICR,- + // +* :, : +, + vv CE-, +* Triton-X+**, 2 0 n NC : NF : HDL RC : E E RA : HDL E RP : +* RPA : AST ALT C ++ 3Tween 2* **3. TBARS Tween, mgml TBARS +* w v +,. +3 Kikugawa, mgml + + -,* mgkg TBARS TBARS NC, NF, RC Tween,* +0 P M ; P M ; P M ; P. 3+3 M ; P/ 223 M +0 mz,21 +1 Hayashi HPLC Yagi +, Protein Quantifi- cation Kit-Rapid / +- Tween ANOVA Tukey-Kramer -* *.. cm / P*.*/ /* ml,. + HPLC ++ *** g, Fig. +. / / P-P + / 2* LC-MS 0 mz + 1.-, / cyanidin 1,+,, C- P+ cyanidin-- -(, - xylopyranosyl-0-glucopyranosyl-galactopyranoside) ;

4 10 /2 +,* Fig. + HPLC chromatogram (/,* nm) of purple carrot anthocyanin extracts P +, cyanidin-- -(, -xylopyranosyl-0 -glucopyranosyl-galactopyranoside) ; P,, cyanidin-- -(, -xylopyranosyl- glucopyranoside) ; P -, cyanidin-- -(, -xylopyranosyl-0 -sinapoyl-glucopyranosyl-galactopyranoside) ; P., cyanidin-- -(, -xylopyranosyl-0 -feruloyl-glucopyranosyl-galactopyranoside) ; P /, cyanidin-- -(, -xylopyranosyl- 0-glucopyranosyl-galactopyranoside). Fig., HPLC chromatogram (A,./* nm ; B,,2* nm) of purple carrot carotenoid extracts P +, a-carotene ; P,, b-carotene ; P -, phytoene. P, cyanidin-- -(, -xylopyranosyl-glucopyranoside) P- P+,2* nm P- cyanidin-- -(, -xylopyranosyl-0-sinapoyl-glucopyranosyl- l,2* nm galactopyranoside) P. cyanidin-- -(, -xylopyranosyl-0- mz M /-0 feruloyl-glucopyranosyl-galactopyranoside) P/ cyanidin- --(, -xylopyranosyl-0-glucopyranosyl-galactopyranoside) ww : bo ww :P + 2*. ;.0.. ; ao,/.* ;,2. 0 P, -+,. ; P -.*0. ; P. +2/. ; P / +0. -, HPLC Fig.,./* nm, P+ P,,2* nm + Fig. -A, B P- -./* nm RC NC, NF bo P, ao max

5 11 : 11 Fig. - E# ects of pigment extracts from purple carrot on plasma corticosterone (A), glucose (B), AST activity (C), and ALT activity (D) in restrained mice NC, no restraint stress/control group ; NF, no restraint stress/fasting group ; RC, restraint stress/ control group ; RP, restraint stress/carotenoid group ; RA, restraint stress/anthocyanin group ; RPA, restraint stress/carotenoidanthocyanin group. Data are meanse. (A) (B) *** P***+., ** P *.*+, * P*.*/, (C) *** P*.**+, (D) *** P*.**+, * P*.*/ RC RP RA RPA NC, NF NC RC NF AST ALT RP RA RPA RC RC Fig.. RA RPA NC NF RC Fig. -C, D AST ALT RA RPA

6 12 /2 +,* Fig. / E# ects of pigment extracts from purple carrot on hepatic triglycerides (A), and total cholesterol (B) in restrained mice Fig.. E# ects of pigment extracts from purple carrot NC, no restraint stress/control group ; NF, no restraint on plasma total cholesterol (A), HDL-cholesterol stress/fasting group ; RC, restraint stress/control group ; (B), and arteriosclerotic index (C) in restrained RP, restraint stress/carotenoid group ; RA, restraint stress/ mice anthocyanin group ; RPA, restraint stress/carotenoid anthocyanin group. Data are meanse. (A) (B) *** P NC, no restraint stress/control group ; NF, no restraint *.**+, ** P*.*+, * P*.*/ stress/fasting group ; RC, restraint stress/control group ; RP, restraint stress/carotenoid group ; RA, restraint stress/ anthocyanin group ; RPA, restraint stress/carotenoid anthocyanin group. Data are meanse. (A) *** P***+., ** P**+., * P**/., (B) ** P**+., * P**/., (C) *** P***+., ** P**+.. RC Fig. / A RP Fig.. A RC NC HDL RP RC RP Fig.. B RA HDL HDL RPA RC Fig. / B RP TBARS RA RPA Fig.. C Fig. 0 TBARS NC NF Fig. / RC

7 13 : 13,- +0,. + bo ao / Fig. 0 E# ects of pigment extracts from purple carrot on lipid peroxidation in plasma (A) and liver tissues (B) in restrained mice NC, no restraint stress/control group ; NF, no restraint stress/fasting group ; RC, restraint stress/control group ; RP, restraint stress/carotenoid group ; RA, restraint stress/ anthocyanin group ; RPA, restraint stress/carotenoid anthocyanin group. Data are meanse. (A) *** P***+., ** P*.*+, * P*.*/, (B) *** P*.**+, * P*.*/ RC RA RPA TBARS RP Fig. 0A RP, RA, RPA RC CoA HMG-CoA TBARS,/ NC NF,0 LDL Fig. 0B,1 LDL,2 delphinidin- --( p-coumaroylrutinoside)-/ -glucoside,3-*

8 14 /2 +,* , / TBARS TBARS,* mgkg TBARS,* mgkg TCH-1,+ TBARS TBARS / bo ao - cyanidin- - -rutinoside, cyanidin- - -galactoside, delphinidin- - - rutinoside + Seyle, H., A syndrome produced by diverse nocuous agents. Nature, +-2, -, ( +3-0)., +, -+--,+,**+ - AGE ,**. / Thernlund, G.M, Ludvigsson, J., Dahlquist, G., Sjoblad, S., Hansson, K., Hagglof, B. and Ivarsson, S.A., Psychological stress and the onset of IDDM in children. Diabetes Care, +2, +-,- +-,3 ( +33/ ). 0 pp. -. /-,**- 1 Kohno, H. and Ohkubo, Y., Comparative glucoregulatory responses on mice to restraint and footshock stress stimuli. Biol. Pharm. Bull.,,+, ( +332).

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食糧　その科学と技術　No.48( )

食糧　その科学と技術　No.48( ) 79 +.* + 1/* A + 80, E + + Bioaccessibility +, 81,. 0* nm bo, Caco-, +* - - 82 - Caco, + Pow +, b-carotene ;,, a-carotene ; -, b-cryptoxanthin ;., lutein ; /, zeaxanthin ; 0, canthaxanthin ; 1, astaxanthin