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- ふさこ うばら
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1 ORYZA CERAMIDE -PTPCDP8TP20CD -WSPWSP8 -LL0.8 -PCPC8PC20 -WSPCWSPC8 -LCLC0.8 Ver.13.0HS
2 ORYZA CERAMIDE Ver.13.0HS NMR 4 ( 1) 1
3 Ver.13.0HS HO CH 2 OH O O OH OH OH C CH CH H 2 NH C O R C H C H (CH 2) 2 C H C H (CH 2) 8 CH 3 R: 1. -(CH 2 ) 7 CH:CHCH 2 CH:CH(CH 2 ) 4 CH (CH 2 ) 14 CH (CH 2 ) 7 CH:CH(CH 2 ) 7 CH (CH 2 ) 16 CH HPLC ( 2 No.7) 3 3 2
4 3. Ver.13.0HS 1884 Thudichum 120 ( ) ( 4) 4060 % ( 5) 4 26% 10% 5 40% 24% 9 ( 6) 3
5 Ver.13.0HS 6 CER 9 ( 7) 5) ( 8) (Lati 4) ) () 4
6 Ver.13.0HS G. Imokawa, J. Invest. Dermatol., 96, 523, Schmelz % Nyberg 5
7 (in vivo) Ver.13.0HS GCFr, 99% -PTFGC 9A TEWL 9B FGC256 mg/kg10%sds TEWL 12 SDS TEWL 9B 9C SDS TEWL GCFrFGC TEWL 9. GCFr -PTFGC TEWL A) SDS B) SDS TEWLC) 12 SDS TEWL n=5-6 6
8 Ver.13.0HS 16 SDS TLC 10 1, 2Cer 1, 2 GlcCerEOS ( 1 )A/B4 GlcCer GCFr 1 EOS, A/B 1SDS GCFr 11 7
9 Ver.13.0HS 10. TLC Cho: FFA: PE: Cer: (190:9:1):10% 8%, GlcCer: (40:12:1), 0.1% 10% 1. GCFr SDS Cho FFA Cer 1 Cer 2 Normal 0.711± ± ± ±0.024 Control 0.669± ± ± ±0.022 GCFr (3 mg/kg) 0.732± ± ± ±0.021 GCFr (10 mg/kg) 0.840± ±0.088* 0.211±0.036* 0.135±0.020 GlcCer (EOS) GlcCer A/B PE Normal 0.035± ± ±0.020 Control 0.034± ± ±0.008 GCFr (3 mg/kg) 0.032± ± ±0.018 GCFr (10 mg/kg) 0.016±0.004* 0.017±0.006* 0.069±0.011 Cho: FFA: PE: CerGlcCer n=5-6*: p<
10 Ver.13.0HS 11. GCFr Cer GlcCer 50 µmcer GlcCer 9
11 Ver.13.0HS 12 KGKG GCFr Control GCFr 10 mg/kg 123,000 SC:SL:SG:KG: 13 GCFr Control GCFr10 mg/kg Control GCFr10 mg/kg 10
12 Ver.13.0HS 14 GCFr Control GCFr10 mg/kg 14300, cornified envelope GCFr Control GCFr10 mg/kg 15. cornified envelope3,000 GCFr 11
13 Ver.13.0HS 16 Western blotting 7 GCSase-GCaseGCFr GCaseGCSase GCFr GCFr GCSase- GCase A) 10% SDS-PAGE Western blotting. B) control 1 12
![Ver.13.0HS 5-1-2 in vitro [GlcCer (d18:2), 3] 3 17Cer 1, Cer 2, GlcCer (EOS) GlcCer A/B 2 17.](/docs-images/94/119054905/images/14-1.jpg "TLC Cho: FFA: Cer:GlcCer:PE: 2. GlcCer (d18:2) Cho FFA Cer 1 Cer 2 (µg/ml) Control - 0.238±0.036 0.053±0.")
14 Ver.13.0HS in vitro [GlcCer (d18:2), 3] 3 17Cer 1, Cer 2, GlcCer (EOS) GlcCer A/B TLC Cho: FFA: Cer:GlcCer:PE: 2. GlcCer (d18:2) Cho FFA Cer 1 Cer 2 (µg/ml) Control ± ± ± ±0.011 GlcCer (d18:2) ± ± ±0.003* 0.071± ± ±0.018* 0.076± ± ± ±0.018* 0.076± ±0.002* GlcCer (EOS) GlcCer A/B PE Control ± ± ±0.017 GlcCer (d18:2) ± ± ± ±0.009* 0.067±0.003** 0.117±0.017* ±0.005* 0.049±0.010* 0.104±0.010 Cho: FFA: PE:Cer GlcCer n=3 *: p<0.05, **: p<
15 Ver.13.0HS GlcCer (d:18:2) 18. GlcCer (d:18:2) Cer GlcCer 50 µmcer GlcCer 14
16 Ver.13.0HS GlcCer (d:18:2) GCase GCSase GlcCer (d:18:2) GCSase-GCaseTGase A) 10% SDS-PAGE Western blotting. B) control 1 15
17 5-2 (in vitro) B16 Ver.13.0HS B16 () 20 (%) g/ (%) C µg/ml B16 ( cells/ml) (60 mm dish) 10% D-MEM 24 () 4 2 2N NaOH (371 ) 450 nm C : 100 % : 95 % 16
18 Ver.13.0HS (melan-a) * 21, 22 * 1 N HCl (70/18 ) n- % of control % of control (µg/ml) (µg/ml) 21. % of control % of control (µg/ml) (µg/ml)
19 Ver.13.0HS 1) (melan-a cells, cells/well) 96well plate 200 nm TPA 10% RPMI ( 95%) 4 1% tritonx-100 PBS (90µlwell) 1 10M -DOPA PBS (10µlwell) (137) 475nm 2) melan-a cells ( cellswell) 90 mm plate 6 well plate 1) 1N NaOH (500µl) (10030 ) 405nm 5-3 (in vitro) Merrill Futerman 1-3) in vitro ) 3% 23. ( 3%) 35 40% 8 18
20 Ver.13.0HS µg/ml (163%) : 1. 95% 2. 95% 3. 95% 4. 95% (300 µg/ml) HS-K 10 %1000 unit/ml 100 µg/ml RITC80-7 (5%CO 2, 37 ) cells (100 µl/well) 1% (FBS) 1000 unit/ml 100 µg/ml RITC µl 3 mg/ml (EtOH 0.5 %) 72 Cell Counting kit nm 5-5 4) 19
21 Ver.13.0HS RBL-2H3 RBL-2H3 3 RBL-2H3 (%) : 1 µg/ml, S.E., n = 6 IgE () RBL-23 ( 1 µg/ml) compound48/80 (ddy, ) compound 48/80 ( 25)compound 48/ compound 48/80 (ddy, ) 4 ( %) 3 3%compound 48/
22 Ver.13.0HS 5-7 () α P. Acnes QOL QOL VISIA Skindex-16 QOL
23 µ Ver.13.0HS 26. *: p<0.05 P. Acnes 27 P. Acnes *: p<
24 Ver.13.0HS Skindex-16 QOL Skindex QOL -100 QOL 30 QOL 30. QOL *: p<
25 Ver.13.0HS ) F344 (5 ) (AOM) 1 2 aberant crypt foci (ACF) mucin depleted foci (MDF) (G 1 CM) 4 23 (G 1 CM initiation ) ACF MDF 1 () 8 ACF MDF 1 45 (G 1 CM post-initiation ) ( 4) PCNA Caspase-3 ( 31) 2 (ACF MDF) 4 AOM PCNA Group Conc. N Number of ACF Number of MDF (ppm) 4 w 8w 4w 8w p< p< p< p< p< p< p< p< p< , , Caspase-3
26 x 200 x 200 PCNA Caspase-3 Ver.13.0HS PCNA Caspase-3 F344 (5 ) 7 32 (AOM 20 mg/kg) 1 2 ACF MDF (ph 2.5) ACF MDF proliferating nuclear antigen (PCNA) cleaved caspase-3 1 : AOM 1 2,3 : 90% (G 1 CM) 100, 250 ppm AOM 1 4 4,5 : AOM 1 2 G 1 CM (100, 250 ppm) 3 6,7 : AOM 250 ppm G 1 CM weeks 1 (24rats) CE-2 2 (24rats) CE-2+100ppm G 1CM CE-2 3 (24rats) CE-2+250ppm G 1CM CE-2 4 (12rats) CE-2 CE-2+100ppm G 1CM 5 (12rats) 6 (9rats) CE-2 CE-2+250ppm G 1CM CE-2+250ppm G 1CM 7 (9rats) CE-2 ; AOM s.c. injection (20mg/kg body weight) ; Sacrifice ( diet 2 ) 32 AOM 25
27 Ver.13.0HS ) NOD/SCID 5 mm (300 mg/kg/day 24 mg/kg/day) 14 ( 33) TUNEL ( 34) NOD/SCID ( 35)
28 Ver.13.0HS p < control glucosylceramide ) Eva-Maria S. et al., J. Nutr., 124 (5), 702 (1994). 2) Merrill A. H. Jr. et al., FASEB, 3A, 469 (1989). 3) Futerman A. H., CHAPTER 4, Current Topics in Membranes, Vol.40, 93 (1994). 4) Elian L. Fragrance Journal, 23 (1), 81 (1995). 5),, 44 (10), 751 (1995). 6) Lena N. et al., J. Nutr. Biochem., 8 (3), 122 (1997). 7) Inamine M. et al., Cancer Sci., 96, (2005). 8) Fujiwara K. et al., Int. J. Clin. Oncol., 16, (2011). 27
29 6. Ver.13.0HS ph ph ph 90% (ph 6.8) 100% 8. 28
30 Ver.13.0HS -PT, PCD () 3050 mg/day 2040 mg/day -P8T () 1119 mg/day mg/day -P20CD () mg/day 36 mg/day -WSP () 3050 mg/day 2040 mg/day -WSP8 () 1119 mg/day mg/day -L () mg/day mg/day L0.8 () mg/day mg/day 9. PT PCD P8T P20CD WSP WSP8 L L0.8 (/100 g) 1.2 g 1.8 g 1.1 g 2.2 g 1.8 g 40.2 g (/100 g) 5.3 g 3.8 g 7.3 g 4.7 g 3.8 g 0.4 g (/100 g) 19.6 g 33.8 g 21.8 g 42.2 g 33.8 g 5.9 g (/100 g) 66.3 g 2.5 g 67.2 g 3.1 g 2.5 g 0.2 g (/100 g) 5.6 g 58.3 g 0.4 g 47.8 g 58.3 g 53.3 g (/100 g) 2.0 g 0.0 g 2.2 g 0.0 g 0.0 g 0.0 g (/100 g) 337 mg 524 mg 328 mg 655 mg 524 mg 52 mg (/100 g) 224kcal 552kcal 232kcal 590kcal 552kcal 268kcal -PT : : : :
31 Ver.13.0HS (447 (456 )) (LD 50 ) 30 g 5 ICR 5000 mg/kg 23±2 50±10% 14 LD mg/kg 3 28 ) 4 Slc:ddy 60 mg/kg ( 60 kg 3.6 g/) % mg/kg 4 ( ) 30
32 Ver.13.0HS -PTPCDP8TP20CD () -WSPWSP8 () -PCPC8PC20 () -WSPCWSPC8 () 1kg -LL0.8 () -LCLC0.8 () 5kg
33 Ver.13.0HS 14. PTP8T PCDP20CD WSPWSP8 LL0.8 () () INCI PCPC8PC20 Cyclodextrin Oryza Sativa (Rice) Bran Oil Glycosphingolipids WSPCWSPC8 Maltosyl Cyclodextrin Cyclodextrin Maltose Oryza Sativa (Rice) Bran Oil Glyco sphingolipids LC 10 Glycerin Water Polyglyceryl 10 Oleate Lecithin Oryza Sativa (Rice) Bran Oil Glycosphingolipids LC Glycerin Water Polyglyceryl 10 Oleate Oryza Sativa (Rice) Bran Oil Glycosphingolipids 32
34 Ver.13.0HS (Oryza sativa Linne) 3.0 % 3.0 % () (HPLC) 5.0 % (1 g1052 ) (1) 10 ppm () Pb (2) 1 ppm ( ) As 2 O g () g ( ) (BGLB ) % 1.13 % % 5.00 % 2.00 % 0.57 % 0.50 % 0.20 % % 1.0% (3.8%) 33
35 Ver.13.0HS (Oryza sativa Linne) 3.0 % 3.0 % () (HPLC ) 8.0 % (1 g1052 ) (1) 10 ppm () Pb (2) 1 ppm ( ) As 2 O g () g ( ) (BGLB ) 40 % 60 % 100 % 34
36 Ver.13.0HS (Oryza sativa Linne) 8.0 % 8.0 % () (HPLC ) 5.0 % (1 g1052 ) (1) 10 ppm () Pb (2) 1 ppm ( ) As 2 O g () g ( ) (BGLB ) % 1.13 % % 5.00 % 2.00 % 0.57 % 0.50 % 0.20 % % 35
37 Ver.13.0HS (Oryza sativa Linne) 20.0 % 20.0 % () (HPLC ) 5.0 % (1 g1052 ) (1) 10 ppm () Pb (2) 1 ppm ( ) As 2 O g () g ( ) (BGLB ) 50 % 50 % 100 % 36
38 Ver.13.0HS (Oryza sativa Linne) 3.0 % 3.0 % () (HPLC ) 5.0 % (1 g1052 ) (1) 10 ppm () Pb (2) 1 ppm ( ) As 2 O g () g ( ) (BGLB ) 40 % 60 % 100 % 37
39 Ver.13.0HS (Oryza sativa Linne) 8.0 % 8.0 % () (HPLC ) 5.0 % (1 g1052 ) (1) 10 ppm () Pb (2) 1 ppm ( ) As 2 O g () g ( ) (BGLB ) 40 % 60 % 100 % 38
40 Ver.13.0HS (Oryza sativa Linne) 0.30 % % () (HPLC ) (1) 10 ppm () Pb (2) 1 ppm ( ) As 2 O g () g ( ) (BGLB ) 40 % 4 % 46 % 10 % 100 % 39
41 Ver.13.0HS (Oryza sativa Linne) 0.80 % % () (HPLC ) (1) 10 ppm () Pb (2) 1 ppm ( ) As 2 O g () g ( ) (BGLB ) 40 % 4 % 46 % 10 % 100 % 40
42 Ver.13.0HS (Oryza sativa Linne) () 3.0 % 0.2 g 2 ml 3.0 % () (HPLC ) 8.0 % (1 g1052 ) (1) 10 ppm ( 2 ) Pb (2) 1 ppm ( 3 ) As 2 O /g () /g ( ) (BGLB ) 60 % 37 % 3 % 100 % 41
43 Ver.13.0HS (Oryza sativa Linne) () 8.0 % 0.2 g 2 ml 8.0 % () (HPLC ) 5.0 % (1 g1052 ) (1) 10 ppm ( 2 ) Pb (2) 1 ppm ( 3 ) As 2 O /g () /g ( ) (BGLB ) 60 % 32 % 8 % 100 % 42
44 Ver.13.0HS (Oryza sativa Linne) () 20.0 % 0.2 g 2 ml 20.0 % () (HPLC ) 5.0 % (1 g1052 ) (1) 10 ppm ( 2 ) Pb (2) 1 ppm ( 3 ) As 2 O /g () /g ( ) (BGLB ) 50 % 30 % 20 % 100 % 43
45 Ver.13.0HS (Oryza sativa Linne) () 3.0 % 0.2 g 2 ml 3.0 % () (HPLC ) 5.0 % (1 g1052 ) (1) 10 ppm ( 2 ) Pb (2) 1 ppm ( 3 ) As 2 O /g () /g ( ) (BGLB ) 60 % 37 % 3 % 100 % 44
46 Ver.13.0HS (Oryza sativa Linne) () 8.0 % 0.2 g 2 ml 8.0 % () (HPLC ) 5.0 % (1 g1052 ) (1) 10 ppm ( 2 ) Pb (2) 1 ppm ( 3 ) As 2 O /g () /g ( ) (BGLB ) 60 % 32 % 8 % 100 % 45
47 Ver.13.0HS (Oryza sativa Linne) () 0.30 % (1) 10 g 50 ml 0.5 g (2) 1 g 5 g 0.5 g 20 ml 2 20 ml 5 ml 10 ml 0.30 % () (HPLC ) (1) 10 ppm ( 2 ) Pb (2) 1 ppm ( 3 ) As 2 O /g () /g ( ) (BGLB ) 40.0 % 38.0 % % 5.0 % 3.7 % 0.3 % % 46
48 Ver.13.0HS (Oryza sativa Linne) () 0.80 % 10 g 50 ml 0.5 g 0.80 % () (HPLC ) (1) 10 ppm ( 2 ) (Pb) (2) 1 ppm ( 3 ) As 2 O /g () /g ( ) (BGLB ) 46.0 % 40.0 % % 3.2 % 0.8 % % 47
49 Ver.13.0HS TEL(0586) () FAX(0586) URL/ OEM F TEL(03) FAX(03) * * * 48
50 Ver.13.0HS 1
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