講演要旨集

(1)

学会創立90周年記念

日本農芸化学会2014年度中四国支部大会

（第 40 回講演会）

日本動物細胞工学会

第33回動物細胞工学シンポジウム

日時：2014年９月26日（金），27日（土）

場所：徳島大学常三島キャンパス

日本農芸化学会中四国支部

(2)

DNA

HBS

(3)

(4)

(5)

(6)

(7)

(8)

(9)

(10)

(11)

(12)

講演会

プログラム

(13)

DNA

(14)

STS

HBS

(15)

PCTK3

STS

1,2 1 1 1 1

2

1 AFI ² STS

Savinin

1 1

D- -2-

D-

Md. Tazul Islam Chowdhury

MOLT-4F

(16)

The synthesis of 4,5-disubstituted-3-deoxy-D-manno-octulosonic acid (Kdo) derivatives

Ruiqin Yi, Tsuyoshi Ichiyanagi Fac. of Agric., Tottori Univ.

Kdo2

1 1

V

1 1 1 1

1 1 1

1

Senna occidentalis L.

1 1

oleuropein aglycon

1 2 2

1 2

-N-Acetylglucosaminidase

1 1

(17)

Screening and characterization of ulvan-degrading bacteria

Sultana Sharmin¹ ¹ ¹

1

O157

1,2 3 1

1 2 3

T-RFLP 18S-rDNA

1 1

rDNA

PA-

RSc0608

(18)

1 1

1 1 2

, ¹ , ²

1 1

, ¹

1 1

1

(19)

a -

1 1

N-

1 1

Pyrococcus horikoshii

¹ ²

1 2

Aeropyrum pernix D-

, ¹, ²,

, ¹ , ²

1 1

1

L- -

1 1

[NiFe]

5'

1 2 2 2

1 2NITE-NBRC

(20)

N-

Click chemistry in vitro in vivo

CDCA3 N-

1 1 1 2

1

1 2

Manoj Pohare

(21)

Association of GSTP1 and ABCC4 polymorphisms with response and toxicity of cyclophosphamide-based chemotherapy on Bangladeshi breast cancer patients

Mohammad Safiqul Islam, Md. Siddiqul Islam, Ferdous Khan¹, Pinky Karim Syeda K. Fatema¹, Abul Hasnat², Kazushige Yokota¹

Dept. Pharm., Noakhali Sci. Technol. Univ., Bangladesh; ¹Dept. Life Sci.

Biotechnol., Shimane Univ.,; ²Dept. Clin. Pharm. Pharmacol. Univ. Dhaka, Bangladesh

Biosynthesis of prostacyclin and its action are up-regulated during the maturation phase of adipocytes

Ferdous Khan, Mohammad Sharifur Rahman, Pinky Karim Syeda K.

Fatema, Mohammad Safiqul Islam, Kohji Nishimura¹, Mitsuo Jisaka, Tsutomu Nagaya, Fumiaki Shono², Kazushige Yokota

Dept. Life Sci. Biotechnol., ¹Center Int. Res. Sci., Shimane Univ., and

2Dept. Clin. Pharm., Tokushima Bunri Univ.

11-Deoxy-11-methylene-prostaglandin (PG) D₂ exerts pro-adipogenic effect on cultured adipocytes during the maturation phase through cellular mechanism different from that of natural PGD₂

Pinky Karim Syeda K. Fatema, Mohammad Safiqul Islam, Ferdous Khan, Kohji Nishimura¹, Mitsuo Jisaka, Tsutomu Nagaya, Fumiaki Shono²,

Kazushige Yokota

Dept. Life Sci. Biotechnol., ¹Center Int. Res. Sci., Shimane Univ., and

2Dept. Clin. Pharm., Tokushima Bunri Univ.

1 2

1

1 1 2

1 2

1 1

(22)

Secoisolarisiresinol

IGF-1

1 1

1

D- SOD

B₁₂

1 2 1 1

1

1 2

B₁₂

, ¹, , ²,

, ¹ ,²

B₁₂

1 1 2

3 1

1 2 3

(23)

B₁₂

(24)

(25)

受賞講演

講演要旨

(26)

2014

Bacillus subtilis ywfE ATP

L- - L-amino acid -ligase (Lal) ¹⁾

Lal Lal

2), 3), 4)

Gln Tyr

AlaGln AlaTyr

Gln Tyr

1/10

1) K. Tabata, H. Ikeda & S. Hashimoto : J. Bacteriol., , 5195-5202 (2005).

2) K. Tabata & S. Hashimoto : Appl. Environ. Microbiol. , 6378-6385 (2007).

3) M. Hayashi & K. Tabata : Appl. Environ. Microbiol. , 3033-3039 (2013).

4) M. Hayashi, K. Tabata, M. Yagasaki & Y. Yonetani : FEMS Microbiol. Lett. , 12-19 (2010).

(27)

2014

DNA

DNA PCR

PCR DNA

Thermus aquaticus DNA Taq DNA PCR

, DNA

3'-5'

DNA 90

3'-5' Pyrococcus furiosus Pfu DNA

Taq DNA PCR

DNA Thermococcus kodakarensis KOD1

P. furiosus

KOD DNA Pfu DNA

80 DNA

DNA

DNA X

DNA Palm Fingers

dNTP KOD DNA

Fingers Palm

dNTP

KOD DNA Taq DNA , Pfu DNA

Taq DNA

2.5 50

DNA 3'-5'

PCR

KOD DNA PCR

KOD DNA 2 PCR

1 KOD DNA PCR KOD

–Plus- Neo Taq DNA 80 1 KOD DNA

DNA

PCR KOD FX Neo Taq DNA

20 20 kb

2013 PCR ,

, PCR KOD SYBR qPCR Mix

KOD DNA

GeneCube

KOD

(28)

2 2014

1.

100 MPa

MPa 2.

3.

JAS

(29)

2014

4 RLYLRIGRR-NH₂, RLRLRIGRR-NH₂,

ALYLAIRRR-NH2, RLLLRIGRR-NH2

MRSA

J. Insect Biotechnol. Sericology, : 25-29.

PS

PS Peptides, : 660-668.

Biosci.

Biotechnol. Biochem., : 683-687.

integrin V

3 cyclic RGD

cyclic RGD cyclic RGD

integrin V 3

Biosci. Biotechnol. Biochem., : 2044-2048.

(30)

2014

EGCg

- -3-

2''-

EGCg EGCg

EGCg

-

6 EGCg

NO

4 2 -

DNA

ATM (Ataxia Telangiectasia) ATM

ATM 2

- ATM

/EGCg

(31)

(32)

シンポジウム

「食と健康」

講演要旨

(33)

900 250

K562

Apigenin-7-glucoside K562

G2/M CD

Chk1 Chk2 Apigenin-7-glucoside (Eur. J. Nutr, 2013, 52, 25 Mol. Nutr. Food Res, 2011, 55, 93)

AD PD

Behavioural Brain Res. 2013, 238, 86

2

J Dermatol Sci,2012, 67, 26 J.Nat.Prod.2009,72, 938)

(34)

EGFP

SIRT1 T2102

SIRT1 T2102 - SIRT1

- T2102 -

T2102 SIRT1 -

T2102

(35)

Diphenylpropane

1936 Scent-Györgyi P

Scent-Györgyi

II

1.5-2

8)

9

1)Kawai et al. J.Biol. Chem. 2008:283:9424-9434, 2)Terao et al. Food Func. 2011:2:11-17, 3) 2010:192:25-32,

4)http://www.naro.affrc.go.jp/project/f_foodpro/subject/a7.html

5)Takumi et al. Food Func. 2012:3:389-398, 6)Murota et al. Arch. Biochem. Biophys.2010:501:91-97, 7)Mukai et al. PLoS ONE 2012:7:e45048, 8)Mukai et al. J. Nutr. 2013:143:1558-1564, 9)Murota et al.

Br. J. Nutr. 2013:109:2147-2153, 10)Nakamura et al. Mol. Nutr. Food Res. 2014:58:310-317.

(36)

30 1984

3 11

1100

8

1980

Caco-2

iPS

M. Shimizu, Interaction between food substances and the intestinal epithelium, Biosci.

Biotechnol. Biochem., , 232-241(2010); FFI , ,

311-318 (2013).

(37)

一般講演

講演要旨

(38)

PCTK3

STS

PCTAIRE kinase 3 cyclin dependent kinase (CDK) Ser/Thr CDK

CDK PCTK3 cyclin A PKA

PCTK3 PCTK3 PCTK3

PCTK3 siRNA HEK293T

PCTK3 phalloidin

F- PCTK3

PCTK3

myosin light chain LIM-domain

kinase PCTK3 RhoA/ROCK

1,2 1 1 1 1 2

1 AFI ² STS

ELISA

CHO

(39)

Savinin

Savinin -benzylidene- -benzyl- -butyrolactone (Procambarous spiculifer)

HL-60 (-)-savinin

savinin

Piperonal (4R)- (4S)-benzyloxazolidinone

(+)- (-)- -benzyl- -butyrolactone savinin

HL-60 Vero WST-8 savinin

HL-60 IC50 10 M

Vero 30 M savinin

1 1

5

Comparative Molecular Field Analysis

CoMFA

CoMFA n-pentyl

(40)

D- -2-

D-

2- D- 6 D- -2-

D- 2- 4A

50 8 Candida antarctica lipase C-6

D- -2-

10 25 2

7

30 7 D-

-2- 100 µM 40

1 µM 120 D- -2-

100 µM 70 10 µM

120

D-

Md. Tazul Islam Chowdhury

D-

1,2,6-

-6-(2- )-D- (dAll-HBI)

D-

51 88 66

70 20 dAll-HBI

10 25 48

10 µM 110 1 mM 107

25 96

100 µM 115 136

(41)

MOLT-4F

D- D- C-3

D- C-6

D- D-

C-5

D- 2- -D- D- L- 4

10~300 M MOLT-4F

37 5 CO2 48

D-

D- 2-

-D- D- L-

The synthesis of 4,5-disubstituted-3-deoxy-D-manno-octulosonic acid (Kdo) derivatives

Ruiqin Yi, Tsuyoshi Ichiyanagi (Fac. of Agric., Tottori Univ.)

Lipopolysaccharides (LPSs) are produced by gram-negative bacteria on their outer membrane and play an important role in the mechanism of bacterial infection. LPS is composed of O-specific polysaccharide, a core oligosaccharide (core OS), and lipid A. The lipid A moiety causes the biological toxicity of LPS; however, the function of a core oligosaccharide of LPS is unclear. Recently, it has been reported that human antibody recognized a core OS of LPS derived from pathogenic strains, so core OS is a focused target for vaccine development. In our development of the core OS synthesis, we have reported a new Kdo intermediate preparing from D-mannose to synthesize the Kdo dissacharides [Kdo ( 2-4 or 2-8)Kdo] in good yield¹.

In this report, the limitation of glycosylation reactions and syntheses of 4,5-branched Kdo trisaccharides using this Kdo disaccharide as an acceptor will be discussed.

1 Ichiyanagi, T. et. al. Tetrahedron, , 67, 5964-5971.

(42)

Kdo2

1 1

LPS

Lipid A Lipid A 2-4 2 3

3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) Kdo

Kdo Kdo Kdo

3-

Kdo 1 Kdo 5

Kdo2

D- Kdo

90% 4 5

4

2

2- -6-

V

1 1 1 1 1

1 1 1

Maoecrystal V ( ) Isodon eriocalyx

6,7-seco-6-nor-15(8->9)-abeo-5,8-epoxy-ent-kaurane [1] HeLa (IC50 = 20 ng/ml)

[2]

Cyclohex-2-en-1-one Danishefsky-

Kitahara Diels-Alder

- 6-exo-

[1] S. H. Li et al., Org. Lett. 2004, , 4327.

[2] J. Gong et al., J. Am. Chem. Soc., , 132, 16745; J. Y. Cha et al., J. Am. Chem. Soc., , 133, 14964; F. Peng et al., J. Am. Chem. Soc., , 134, 18860; P. Lu et al., J. Am. Chem. Soc., , 135, 14552.

(43)

1 1

2

SPME GC-MS 2- 25.77%

DPPH

TLC

Fr. 1 5 Fr. 1 4

Fr. 4 Fr. 1 4

80°C

4 oleuropein aglycon

1 2 2 ,

1 2

EtOH EtOAc

oleuropein aglycon A B

B oleuropein DPPH

ORAC 2

DPPH AANa ORAC E

Trolox 7 9

HORAC

ESR oleuropein ORAC

HORAC

ESR HORAC ORAC

OH ESR O2 OH C2H5

oleuropein AANa Trolox

DPPH ORAC

HORAC ESR oleuropein OH

(44)

-N-Acetylglucosaminidase

1 1

-N-acetylglucosaminidase (GlcNAcase)

Paecilomyces sp. F13 Paecilomyces sp. F2281 Paecilomyces sp. F30 Pochonia suchlasporia var.

suchlasporia TAMA87 F40 4

GlcNAcase

4 F30 F40

2

2 MeOH

MeOH

2

MeOH MeOH

MeOH

F30 F40

(45)

Screening and characterization of ulvan-degrading bacteria

Ulvans are complex sulfated polysaccharides in the cell walls of green algae belonging to the genus Ulva. The objective of this study is to discover ulvan-degrading bacteria, which can contribute to potential use of ulvan oligomers or monomers. Ulvan-degrading bacteria were screened on agar plates with purified ulvan as a sole carbon source in artificial seawater and visualized with cetylpyridinium chloride (CPC). Bacteria samples were collected from feces of the molluscs living on the surface of green algae, Ulva ohnoi. Ulvan-degrading enzymes were monitored from culture supernatants of screened bacteria by using ulvan gel and CPC. Finally, one strain KUL42 was isolated as an ulvan-degrading bacterium. KUL42 was classified as Alteromonas sp. by 16S rRNA gene sequence. The crude enzymes were purified by ammonium sulfate precipitation from the supernatant of bacterial culture grown with ulvan, followed by anion exchange and hydrophobic interaction chromatographies. With CPC method, two proteins with molecular masses of 45k and 60k Daltons were indicated to have activities. N-terminal amino acid sequences of these proteins were determined. Corresponding genes for these proteins were

blast-searched using draft genome sequence of KUL42. As a result, the 45kD- and 60kD-proteins were identified as sulfatase and -xylosidase, respectively. Sulfatase is a enzyme to remove sulfate group from substrates. The 60kD-protein is classified into glycoside hydrolase family 43.

Sultana Sharmin¹ ¹ ¹

1

Fe²⁺

pH 2 Acidithiobacillus ferrooxidans Fe²⁺ Fe³⁺ CaCO3

pH 4

DNA PCR 16S rRNA

DGGE

PCR

DGGE Ferrovum myxofaciens 95%

UMEÅ 9K FS APPW

Ferrovum pH 10 mM

20 A.

ferrooxidans Gallionella A. ferrooxidans ATCC23270

10 mM Ferrovum 10 mM

Ferrovum Ferrovum A. ferrooxidans

(46)

O157

1,2 3

1 2 3

O157

O157sakai

25 30

1/10 LB

0.01 ppm 25 5 99%

O157

T-RFLP 18S-rDNA

1 1

T-RFLP SSU (16S-rDNA 18S-rDNA)

T-RFLP 4

T-RF

16S-rDNA T-RFLP

18S-rDNA T-RFLP

T-RFLP 18S -rDNA V4-5 V7-8 2

30 cm 10

5 cm 10 cm 3 ISOIL for Beads Beating DNA

±1 4

V4-5 V7-8 GeneMapper

16S-rDNA Msp Hha

Xsp T-RFLP

V4-7(F-565-R-1631) PCR T-RFLP

(47)

DNA Pot1

Pot1

pot1 Exo1

Rqh1 Exo1 Rqh1

rDNA

Pot1 DNA Pot1

Rqh1 rqh1-hd pot1 rqh1-hd

DNA M

SAC SAC

pot1 rqh1-hd SAC

SAC pot1 rqh1-hd

Mad2 Bub1

pot1 rqh1-hd SAC

SAC rDNA

SAC

(48)

PA

Rob1 Rob2

Rhomboid Rob1 Rob2

PomBase 2 C

SPAC57A7.09 SPAC57A7.09 PA

Protease Association Domain C

1⁺ PA

Rob1 Rob2 Pzr1

PA-

Pzr1 Rob1 Rob2 rob1 rob2

Pzr1-GFP

rob2 GFP 27 kDa rob1

Rob1 Pzr1 Pzr1-GFP

Pzr1

RSc0608

RSc0608 Saccharomyces cerevisiae

(49)

1 1

( )

C12K

C14K C18:2K 350 mM pH 10.5

100 g 5 g 1.7 g

1.7 g 68 mL 500

C12K

10% C18:2K 10%

C14K 5% 10%

10% 17%

1 1

Lc. lactis

Leu. citreum Leu. citreum

(50)

1 1 2

1 2

Penicillium Penicillium

Penicillium pinophilum NBRC 6345

C4K C6K C8K C10K

C12K C14K C18:1K

C18:2K C18:3K

C10K P. pinophilum 10 4

9 P.

pinophilum C10K MIC 175 mM

LAS

OPP-Na MIC 87.5 mM C10K

C10K

1 1

1

Aspergillus kawachii - - A

-A

A -A A -A

A -A

Aspergillus kawachii NBRC 4308

SLS 30 200 rpm 3

A -A

A -A K2HPO4

1.0 g /L 690 U/g-substrate

400 U/g-substrate

(51)

1

, ¹

D- D-

1 0.1% +3%

28 2

3 PDB

C2

1 1

1

Zymomonas mobilis

38 Z. mobilis TISTR548

40 Z. mobilis 200M

Z. mobilis 200M 5 µg/ml 100 µg/ml

YPD 240

3 41

Z. mobilis 200M Z. mobilis TISTR548

(52)

a -

1 1

1,3 1,2 M3FX

N- N- M3FX a

Gal 1-3[Fuc 1-4]GlcNAc 1- N- N-

N- 100%

N-

1,3Fuc’ase Gb 2.8 kg 1.3 kg

0.2 M NaCl 20 mM Tris-HCl pH8.0

DEAE QA SP

- 1,3Fuc’ase Gb

a PA G2F2GN2M3FX SF-HPLC

SDS-PAGE 1,3Fuc’ase Gb 140 kDa

1,3Fuc’ase

58 kDa 1,3Fuc’ase Gb

N-

1 1

µM N- FNGs

FNGs

FNGs GlcNAc “GN1 ”

FNGs N-

FNGs

GN1-FNGs N-

FNGs

FNGs FNGs PA Con A

Con A -Fuc’ase -Gal’ase -GlcNAc’ase

GN1-FNGs

FNGs ,

(53)

Pyrococcus horikoshii

1 2

HSDH /

HSDH

HSDH Pyrococcus horikoshii

HSDH

20 % MPD 100 mM pH 6.2

NW12 2.3

NADP(H)

NADP NADP (Ki

11.5 nM) NADP(H) C2 R40 K57

NADP P. horikoshii HSDH NADP

NADP

Aeropyrum pernix D-

1 2

D- Dye-DLDH FAD

D-

D- D-

Sulfolobus tokodaii Dye-DLDH

S. tokodaii DLDH

Aeropyrum pernix ORF APE_0487 X

pET-15b/ APE_0487 Rosetta-gamiTM 2 (DE3) 80 ,

10 min 3 SDS-PAGE

Hg

MAD 1.9 DALI

Rhodopsedomonas palustris Putative dehydrogenase ( 3PM9 )

(54)

1 1 1

3

Superdex 200 Phenyl Sepharose CL-4B SDS-PAGE 32 kDa

32 kDa D- SUL-I N

SUL-IA SUL-IA T

SUL-IA D- SUL-IA T

1 T Th1 IFN- 2 T

Th2 IL-4 SUL-IA T

Th1

L- -

1 1

NAD NadB L-

LAO AspDH

ImAsp

144 NadB 2

QS, NadA ImAsp QS

DHAP LAO AspDH, QS X

LAO:QS

Pyrococcus horikoshii LAO QS pET Duet

DEAE- 4 LAO:QS

QS Fe-S LAO, QS

(55)

[NiFe]

H2ase H2ase

MC4100 HyaAB 2

[NiFe]H2ase HyaAB

HyaAB

HyaAB HyaA C His

hyaAB(SC-His6) pUC19 HyaB

N StrepTagII hyaAB (LN-STII) pUC19

MC4100 hyaAB

HyaAB Native-PAGE

HyaAB(SC-His6) HyaAB (LN-STII)

5'

NITE-NBRC

[ ]

5

[ ]

Streptomyces mobaraensis

- Sm-ELA Sm-ela 5 1

3 5

mRNA

(56)

N-

N -

N N -

Wnt10a N -

N -

Wnt10a N - COS-1

N -

N - Glycopeptidase F N -

N -

N - N -

Wnt10a

N - N N -

N- RI

RI RI

Click chemistry in vitro

in vivo Click chemistry

in vitro in vivo

Click chemistry SDS-PAGE

in vitro in vivo N-

N- N Gly

in vitro in vivo

RI 1

SDS-PAGE Cys Ser

(57)

CDCA3 N-

N- N

N- CDCA3

Swiss-Prot 43,000

N- N-

CDCA3

cDNA N- G2A

[³H]

CDCA3 N- N-

CDCA3 C EGFP

CDCA3-EGFP G2A N-

CDCA3 CDCA3 N-

1 1 1 2 1

1 2

Arabis flagellosa

cDNA

10 M 18 1 mM

ZnSO4 CdSO4 RNA

dT cDNA 24

cDNA pCR4-TOPO

PCR BLAST

A. thaliana VSP1

LOX2 jacalin lectin (JL) family protein

A. halleri plant defensin (PDF1.3) zinc finger family protein

VSP1 PDF1.3 48

65 75

(58)

Manoj Pohare

N

in vitro

(59)

Association of GSTP1 and ABCC4 polymorphisms with response and toxicity of cyclophosphamide-based chemotherapy on Bangladeshi breast cancer patients

Mohammad Safiqul Islam, Md. Siddiqul Islam, Ferdous Khan¹, Pinky Karim Syeda K. Fatema¹, Abul Hasnat², Kazushige Yokota¹(Dept. Pharm., Noakhali Sci.

Technol. Univ., Bangladesh; ¹Dept. Life Sci. Biotechnol., Shimane Univ.,; ²Dept.

Clin. Pharm. Pharmacol. Univ. Dhaka, Bangladesh)

Cyclophosphamide (CPA) used in breast cancer along with epirubicin and 5-fluorouracil is transported by ABCC transporters and detoxified by glutathione S-transferases (GST). The activities of these enzymes and transporters may vary due to the presence of genetic polymorphisms. The effects of GSTP1rs1695 and ABCC4rs9561778 polymorphisms on the response and toxicities of CPA-based chemotherapy were evaluated on Bangladeshi breast cancer patients. Total 256 patients with invasive breast cancers were recruited of which 117 patients received neoadjuvant chemotherapy to examine the response as well as toxicities and another 139 patients received adjuvant chemotherapy to evaluate only the toxicities. Genetic polymorphisms were detected by using polymerase chain reaction-restriction fragment length polymorphism. Patients carrying AG, GG and AG plus GG genotypes of GSTP1rs1695 were more likely to have good response whereas no association of ABCC4rs9561778 was found with the chemotherapy response. Patients carrying GT and GT plus TT genotypes of ABCC4rs9561778 were found to be associated with anemia, neutropenia, leukopenia and gastrointestinal toxicities when compared with GG genotype whereas no association was found with thrombocytopenia. GSTP1rs1695 was not associated with any type of toxicities investigated.

Biosynthesis of prostacyclin and its action are up-regulated during the maturation phase of adipocytes

Ferdous Khan, Mohammad Sharifur Rahman, Pinky Karim Syeda K. Fatema, Mohammad Safiqul Islam, Kohji Nishimura¹, Mitsuo Jisaka, Tsutomu Nagaya, Fumiaki Shono², Kazushige Yokota (Dept. Life Sci. Biotechnol., ¹Center Int. Res.

Sci., Shimane Univ., ²Dept. Clin. Pharm., Tokushima Bunri Univ.)

Prostacyclin alternatively termed prostaglandin (PG)I2 is an unstable metabolite synthesized by the arachidonate cyclooxygenase pathway and rapidly hydrolyzed to 6-keto-PGF1 , a stable inactive product. Earlier studies have reported that prostacyclin analogues are capable of stimulating adipose differentiation under certain culture conditions. However, biosynthesis of prostacyclin has not been determined comprehensively at different life stages of cultured adipocytes. In this study, we monitored the endogenous synthesis of prostacyclin in cultured 3T3-L1 cells by the quantification of 6-keto-PGF1 by its specific immunological assay. The cultured adipocytes after 4-6 days of the maturation phase were found to have the highest capability to synthesize endogenous prostacyclin, which was accompanied by enhanced gene expression of PGI synthase and the IP receptor for prostacyclin.

Selective IP agonists effectively rescued the accumulation of fats inhibited in the presence of aspirin, a general cyclooxygenase inhibitor. On the other hand, specific antagonists for the IP receptors appreciably blocked the storage of fats during the maturation phase. Taken together, our results indicate that biosynthesis of prostacyclin and its action are closely linked with up-regulated adipogenesis during the maturation phase of adipocytes.

(60)

11-Deoxy-11-methylene-prostaglandin (PG) D₂ exerts pro-adipogenic effect on cultured adipocytes during the maturation phase through cellular mechanism different from that of natural PGD2

Pinky Karim Syeda K. Fatema, Mohammad Safiqul Islam, Ferdous Khan, Kohji Nishimura¹, Mitsuo Jisaka, Tsutomu Nagaya, Fumiaki Shono², Kazushige Yokota (Dept. Life Sci. Biotechnol., ¹Center Int. Res. Sci., Shimane Univ., ²Dept. Clin.

Pharm., Tokushima Bunri Univ.)

Prostaglandin (PG) D2 and its dehydration products, PG of J2 series, are known to serve as pro-adipogenic factors in adipocyte differentiation. Here, we studied the effect of 11-deoxy-11-methylene-PGD2 (11dm-PGD2), a chemically stable, isosteric analogue of PGD2, on the accumulation of fats during the maturation phase of cultured adipocytes and compared its efficacy with that of parent PGD2. The dose-dependent analysis of those effects revealed that 11dm-PGD2 was more potent than PGD2 in rescuing the storage of fats attenuated in the presence of indomethacin. Gene expression analysis confirmed the constitutive expression of two types of cell-surface receptors for PGD2 including the DP1 receptor and DP2/CRTH2 receptor. Selective agonists for either of the DP1 receptor and the CRTH2 receptor were able to enhance adipogenesis suppressed in the presence of indomethacin to a similar extent. On the other hand, a selective antagonist for the DP1 receptor more preferentially blocked the pro-adipogenic effect of PGD2 than that of 11dm-PGD2 while a selective antagonist for the CRTH2 receptor was more inhibitory for the action of 11dm-PGD2 as compared with PGD2. These findings indicate that 11dm-PGD2

stimulates adipogenesis through cellular mechanism different from that of natural PGD2.

1 2

PMI-PUFA 20:3 -5 11 14

20:4 -5 11 14 17 PMI-PUFA

C20PMI-PUFA -

PMI-PUFA 24

20:2 -11 14 CHO-K1

- CHO

(HepG2 MKN74) MKN74

50%

C18PUFA C20PMI-PUFA

(61)

1

1 1 2

1 2

1 LPA

LPA LPA

Perio paper

1 L paper 10 mM Bligh&Dyer

LC-MS/MS LPA

LPA 18 1 1

CT LPA

LPA

1 1

20% HFD

C57BL/6J 5 HFD HFD

HFD HFD HFD HFD 7

11

TG HDL- NEFA

HFD HDL-

(62)

Secoisolarisiresinol

Secoisolarisiresinol(SECO) 7 C57BL/6 SECO

Secoisolariciresinol diglucoside(SDG) 1.0% DIO 4

SECO SECO

6 SD DIO

400(PEG400) =50 50 50%

SECO 6 mg/kgBW 1 1 4

AIN93G SECO 6 mg/kgBW DIO

7 C57BL/6NJcl DIO 50%

SECO 6 mg/kgBW 1 1 4

IGF-1

1 1

1

APP APP

2 APP

IGF-1 1

IGF-1

IGF-1 APP

5 SD APP 7

IGF-1 APP

APP MuRF1 Myostatin

mTOR IGF-1

IGF-1 IGF-1

(63)

-

D- SOD

D- C-3 D- Caenorhabditis

elegans

sod-3 sod-3

D- sod-3

(GFP)

SOD-3::GFP CF1553 (muIs84)

D- 20

10 20

ImageJ sod-3 D- sod-3

D- sod-3

(64)

B₁₂

1 2 1 1 1

1 2

B12 (B12)

B12 B12

B12

B12 B12

B12

0.1 %, 0.2 %

0.5 % (w/w) 25°C 3

B12 Lactobacillus delbrueckii subsp.

lactis ATCC 7830 B12 E. coli 215

B12

MII-12 62.2 14.5 ng/g B12 B12 B12

MII-12 0.5 % B12 0.3

g/100g 3 B12 B12

B12

1 2

2

B12(B12) B12 B12[c-lactone]

B12 B12

B12

L.delbrueckii subsp. lactis ATCC7830

B12 6.0 ~ 18.7 g/100 g 7.0 g/100 g

E. coli 215 TLC LC/ESI-MS/MS

B12

(65)

B₁₂

1 1 2 3

B12(B12) B12

B12 B12

B12

Lactobacillus delbrueckii subsp. lactis ATCC7830 B12 E. coli

215 LC/ESI-MS/MS

HPLC

19 15 B12

LC/ESI-MS/MS B12 B12

15 8 192 415 g/100 g B12

B12 15 4

B12

B12 B12

B12[c-lactone] B12[c-lactone] B12

T

T B12

10 30 ppm B12

10 M 180

Sep-Pak C18 HPLC

10 ppm B12 20

B12 361 nm 364 nm 520 – 550 nm 550

– 585 nm HPLC 60 2

30 ppm B12

2

B12 2

(66)

(67)

140725̲AD̲JAACT シンポジウム広告̲入稿.pdf 2 14/07/25 10:40

(68)

2014年度日本農芸化学会中四国支部大会

（第40回講演会）

実行委員長：大政健史

連絡先：徳島大学大学院ソシオテクノサイエンス研究部 T E L：088-656-7408

E-mail：[email protected] 2014年度日本農芸化学会中四国支部大会実行委員会

実行委員長大政健史（徳島大院・STS研究部）

実行副委員長田中保（徳島大院・HBS研究部）

総務・会計・広報

○向井理恵（徳島大院・HBS研究部），湯浅恵造（徳島大院・STS研究部）

要旨集・プログラム

○近藤美樹（徳島文理大），間世田英明（徳島大院・STS研究部），

兼目裕充（徳島文理大），櫻谷英治（徳島大院・STS研究部）

会場

○金丸芳（徳島大院・SAS研究部），中村光裕（徳島大院・SAS研究部），

中川秀幸（徳島大院・SAS研究部），林直宏（大塚化学（株）），

岡崎貴世（四国大），新居佳孝（徳島県立工業技術セ）

懇親会

○増田俊哉（徳島大院・SAS研究部），横井川久己男（徳島大院・SAS研究部），

寺尾純二（徳島大院・HBS研究部），川上竜巳（徳島大院・SAS研究部），

酒井徹（徳島大院・HBS研究部）

(69)

支部からのお知らせ

1. 学会創立90周年記念第41回講演会（支部例会）

開催日：2015年１月24日（土）

場所：水産大学校

内容：受賞講演，一般講演

講演申込締切：2014年12月22日（月）

講演要旨締切：2015年１月５日（月）

世話人：原田和樹（水産大学校）

2. 学会創立90周年記念第24回市民フォーラム開催日：2014年11月８日（土）

場所：愛媛大学内容：招待講演

世話人：菅原卓也（愛媛大学農学部）

3. 学会創立90周年記念第18回若手研究者シンポジウム開催日：2014年９月20日（土）

場所：愛媛大学農学部内容：招待講演

世話人：渡辺誠也（愛媛大学農学部）

日本農芸化学会中四国支部事務局

〒700-8530 岡山市北区津島中 1-1-1 岡山大学大学院環境生命科学研究科農生命科学専攻生物機能化学講座内

支部ホームページ：http://jsbba-cs.jp

講 演 要 旨 集 - 日本農芸化学会中四国支部