メイラード反応に関する研究

東北⼤学⼤学院 農学研究科 機能分子解析学分野教授 宮澤 陽夫

アミノリン脂質のメイラード反応の分析食品とヒト血液における反応と⽣理機能

deoxy-D-fructosyl phosphatidylethanolamineの分析脂質の後期糖化産物の同定と機能

食品の糖化抑制成分高血糖障害への関与の解明

メイラード反応に関する論⽂・著書・総説のリスト

1. N. Shoji, K. Nakagawa, A. Asai, I. Fujita, A. Hashiura, Y. Nakajima, S. Oikawa, T. Miyazawa, LC-MS/MS analysis of carboxymethylated and carboxyethylated phosphatidylethanolamines in human erythrocytes and blood plasma, J. Lipid Res., in press (2010)

2. T. Miyazawa, D. Ibusuki, S. Yamashita, K. Nakagawa, Analysis of Amadori-glycated phosphatidylethanolamine in the plasma of healthy subjects and diabetic patients by liquid chromatography-tandem mass spectrometry. Ann. N. Y. Acad. Sci., 1126, 291-294 (2008)

3. K. Nakagawa, D. Ibusuki, S. Yamashita, T. Miyazawa, Glycation of plasma lipoprotein lipid membrane and screening for lipid glycation inhibitor. Ann. N. Y. Acad. Sci., 1126, 288-290 (2008)

4. 宮澤陽夫, 仲川清隆, 庄子真樹, ⽣体膜脂質グリケーションの実態と阻害成分, アンチエイジング･ヘルスフード（水島裕 監修）, サイエンスフォーラム, pp. 135-143 (2008)

メイラード反応に関する論⽂・著書・総説のリスト

5. T. Miyazawa, Do green tea polyphenols inhibit protein glycation? IMARS Highlights (Research Commentaries for Member of The International Maillard Reaction Society), 2, 6 (2007)

6. T. Miyazawa, Suppressed colonization of Helicobacter pylori by food protein-derived melanoidins, IMARS Highlights (Research Commentaries for Member of The International Maillard Reaction Society), 2, 11 (2007)

7. S. Yamashita, T. Miyazawa, Utilization of various receptors for advanced glycation end products as biomarkers for disease prevention, IMARS Highlights (Research Commentaries for Member of The International Maillard Reaction Society), 2, 8 (2007)

8. S. Yamashita, T. Miyazawa, Do advanced glycation end products trigger Alzheimer’s disease? IMARS Highlights (Research Commentaries for Member of The International Maillard Reaction Society), 2, 15 (2007)

メイラード反応に関する論⽂・著書・総説のリスト

9. O. Higuchi, K. Nakagawa, T. Tsuzuki, T. Suzuki, S. Oikawa, T. Miyazawa, Aminophospholipid glycation and its inhibitor screening system: a new role of pyridoxal 5’-phosphate as the inhibitor. J. Lipid Res., 47, 964-974 (2006)

10. 宮澤陽夫, 庄子真樹, 仲川清隆, ⽣体膜脂質グリケーションの証明. 分析化学, 55, 907-917 (2006)

11. K. Nakagawa, J.H. Oak, O. Higuchi, T. Tsuzuki, S. Oikawa, H. Otani, M. Mune, H. Cai, T. Miyazawa, Ion trap tandem mass spectrometric analysis of Amadori-glycated phosphatidylethanolamine in human plasma with or without diabetes. J. Lipid Res., 46, 2514-2524 (2005)

12. Y. Tokita, Y. Hirayama, A. Sekikawa, H. Kotake, T. Toyota, T. Miyazawa, T. Sawai, S. Oikawa, Fructose ingestion enhances atherosclerosis and deposition of advanced glycated end-products in cholesterol-fed rabbits. J. Atheroscler. Thromb., 12, 260-267(2005)

メイラード反応に関する論⽂・著書・総説のリスト

13. T. Miyazawa, J.H. Oak, K. Nakagawa, A convenient method for preparation of high-purity Amadori-glycated phosphatidylethanolamine and its prooxidant effect. Ann. New York Acad. Sci., 1043, 276-279 (2005)

14. T. Miyazawa, J.H. Oak, K. Nakagawa, Tandem mass spectrometry analysis of Amadori-glycated phosphatidylethanolamine in human plasma. Ann. New York Acad. Sci.,1043, 280-283 (2005)

15. K. Nakagawa, J.H. Oak, T. Miyazawa, Angiogenic potency of Amadori-glycated phosphatidylethanolamine. Ann. New York Acad. Sci., 1043, 413-416 (2005)

16. J.H. Oak, K. Nakagawa, S. Oikawa, T. Miyazawa, Amadori-glycated phosphatidylethanolamine induces angiogenic differentiations in cultured human umbilical vein endothelial cells. FEBS Lett., 555, 419-423 (2003)

メイラード反応に関する論⽂・著書・総説のリスト

17. J.H. Oak, K. Nakagawa, T. Miyazawa, UV analysis of Amadori-glycated phosphatidylethanolamine in foods and biological samples. J. Lipid Res., 43, 523-529 (2002)

18. S. Lertsiri, J.H. Oak, K.Nakagawa, T. Miyazawa, The occurrence of a novel hydrophilic hydroperoxide, 3-hydroxy-5-hydroperoxy-2-methyl 5,6-dihydropyran-4-one, as a cytotoxic glycation product in human plasma. Biochim. Biophys. Acta, 1573, 48-54 (2002)

19. 玉 正浩, 仲川清隆, 宮尾興平, 並木満夫, 宮澤陽夫, リン脂質アマドリ化合物の⽣成と細胞毒性に対するタマネギ抽出物の抑制効果. 日本食品科学工学会誌, 49 (10), 646-651 (2002)

20. J. Oak, K. Nakagawa, T. Miyazawa, Synthetically prepared Aamadori-glycated phosphatidylethanolaminecan trigger lipid peroxidation via free radical reactions. FEBS Letters., 481, 26-30 (2000)

メイラード反応に関する論⽂・著書・総説のリスト

21. S. Lertsiri, M. Shiraishi, T. Miyazawa, Identification of deoxy-D-fructosyl phosphotidylethanolamine as a non-enzymic glycation products of phosphatidylethanolamine and its occurrence in human blood plasma and red blood cells. Biosci. Biotech. Biochem., 62, 893-901 (1998)

LC-MS/MSによるヒト血漿のアマドリ型脂質の分析Analysis of Maillard Reaction Products in Plasma

with LC-MS/MS

Laboratory of Food and Biodynamic ChemistryGraduate School of Agricultural Science, Tohoku University

Prof. Teruo Miyazawa

Laboratory of Food and Biodynamic ChemistryGraduate School of Agricultural Science, Tohoku University

Prof. Teruo Miyazawa

O

H2N O PO

OHO

H OO

O

R1

R2

O

HOHO

OH

OHOH

N O PO

OHO

H OO

R1

R2

O

OH

HOHO

OH

CHO

OH

+

2OOH

OHOH OH

CH

O

HN O PO

OHO

H OO

O

R1

R2

Phosphatidylethanolamine (PE)

Glucose

Schiff base

Amadori-glycated phosphatidylethanolamine(Amadori-PE)

A. Ravandi et al, FEBS Lett. (1996)R. Bucala et al, Proc. Natl. Acad. Sci. USA (1993)

M.O. Lederer et al, Carbohydr. Res. (1997) S. Lertsiri et al, Biosci. Biotech. Biochem. (1998) J.H. Oak et al, FEBS Lett. (2000)J.H. Oak et al, J. Lipid Res. (2002)J.H. Oak et al, FEBS Lett. (2003)

Lipid Glycation - 脂質の糖化Lipid Glycation - 脂質の糖化

PE standard (744.6, 18:1/18:1)

H2N O PO

OHO

H

141

OO

OR1

R2O

603

M.W. 744PEO

OH

OHOH OH

CH2 HN O

OPO

OHO O

OR1

R2O

H

303

603

M.W. 906Amadori-PE

Neutral Loss Scan Mode303 141

4000Q TRAPTM LC/MS/MS(Applied Biosystems Ltd.)

MW 906 MW 744

M.W. M.W.

+Na+Na

Amadori-PE standard (906.6, 18:1/18:1)

Qualitative Analysis of Authentic Amadori-PE and PE by Tandem MS Neutral-Loss Scan ModeQualitative Analysis of Authentic Amadori-PE and PE by Tandem MS Neutral-Loss Scan Mode

HbA1c(%); 7.2, Age; 72, women

2OOH

OHOH OH

CH

O

HN O PO

OHO

H OO

O

R1

R2

2OOH

OHOH OH

CH

O

HN O PO

OHO

H OO

O

R1

R2303

Plasma Glycated Phospholipids of Patients with DiabetesPlasma Glycated Phospholipids of Patients with Diabetes

MS/MS

4000Q TRAPTM LC/MS/MS (Applied Biosystems Ltd.)

R1, R2 = 18:1

OOH

OHOH OH

CH2 HN O

OPO

OHO O

OR1

R2

O

H303 603

MW 906

MS/MS

H2N O PO

OHO

H

141O

OO

R1

R2

O

MW 744PE

Parent ionFragment ion

Amadori-PE

R1, R2 = 18:1

Fragment ion

Parent ion

906603

603

603

744

Neutral loss -303

Neutral loss -141

Tandem MS of Authentic Amadori-PE and PE Tandem MS of Authentic Amadori-PE and PE

アミノリン脂質のメイラード反応とその阻害物質アミノリン脂質のメイラード反応とその阻害物質Maillard Reaction of Aminophospholipid and its InhibitorsMaillard Reaction of Aminophospholipid and its Inhibitors

Maillard reaction: アミノ化合物とカルボニル化合物の非酵素的褐変反応

Glucose Schiff base Amadori productAmadori

rearrangement

ProteinAmino acid

L. C. Maillard. / Hebd. Seances. Acad. Sci. 154 (1912)

アミノリン脂質のメイラード反応に着目

R. Bucala. / Proc. Natl. Acad. Sci. U.S.A. 90 (1993)

AminophospholipidO

O

O

O

HOPO

O

OH

HNO

OH

OHHO

OH

S. Lertsiri. / Biosci. Biotechnol. Biochem. 62 (1998)Amadori-PE

Maillard Reaction (メイラード反応)Maillard Reaction (メイラード反応)

O

O

O

O

HOPO

O

OH

HNO

OH

OHHO

OH

Angiogenesis

FEBS Lett. 555 (2003)

Lipid peroxidation

FEBS Lett. 481 (2000)Aged rat plasmaFoods,

UV labelingHuman plasmaLC/MS/MS

J.H. Oak., et al. K. Nakagawa., et al.

99.98 % 0.02 %

OH

OHOH

OH

CH2OH

CO

H

D-Glucose

O

OHOH

OH

CH2OH

H, OH

+

O

O

O

O

HOPO

O

OHH2N

Phosphatidylethanolamine (PE)

O

O

O

O

HOPO

O

OHN

HO

OH

HO CH2OH

HO

Schiff-PEAmadori

rearrangement

J. Lipid Res. 43 (2002)J. Lipid Res. in press (2005)

Amadori-PE

J.H. Oak., et al. J.H. Oak., et al.

Amadori-PEは高血糖をともなう疾病の増悪化に関与

Glycation of AminophospholipidGlycation of Aminophospholipid

HbA1c (%) of diabetic patients

100

50

0Phos

phat

idyl

chol

ine

Hyd

rope

roxi

de

Normal n=11

<6.0n=5

6.0-6.5n=15

6.5-7.0n=8

>7.5n=19

7.0-7.5n=15

Mean±S.D.

* **

T. Nagashima. and T. Miyazawa., et al./ Diabetes Res. Clin. Pract. 56 (2002)

(%)

(Rel

ativ

e in

tens

ity)

Red blood cellsHemoglobin (Hb) Glucose HbA1c

+

脂質とタンパク質のグリケーションはどちらが重要なのか？

H.F. Bunn, et al./ J. Biol. Chem. 254 (1979)

Glycation and Lipid Peroxidation in vivoGlycation and Lipid Peroxidation in vivo

LDL Density 1.019-1.063MW 2-3x106

Diameter 19-22 nm

ApoB

PC, PETG

Cholesterol

Cholesterolester

Lipids 77%Triglyceride 10%Cholesterol ester 37%Free-cholesterol 8%Phospholipids 22%

ApoB 98%Protein 23%

Phosphatidylcholine 13%Sphingomyelin 6%Phosphatidylethanolamine 2%Lysophosphatidylcholine 1%

LDL

S. Benitez et al. / Atherosclerosis 177 (2004)

脂質グリケーションとタンパク質グリケーションの速度⽐較にLDLを用いた

Low-density Lipoprotein (LDL)Low-density Lipoprotein (LDL)

Lipids extractionby Folch method

Incubation in PBS (37oC, 0-72 h)

LipidsProteins

Scintillation counter

LDL(2 mg/mL protein)

D-Glucose(30 mM)

D-Glucose-1-[14C]

O

OHHO

OHHO

HOO

OHHO

OHHO

HO

Phospholipids (HPLC-ELSD)PhosphatidylethanolaminePhosphatidylcholineSphingomyelinLysophosphatidylcholine

ApoB Protein (ELISA)Neutral lipids (Enzyme)Total cholesterolFree cholesterolTriglyceride

Lipid hydroperoxides (FOX assay)Total LOOH

MethodologyMethodology

ApoB (515563 Da)

PE (744 Da)

グリケーションを受ける脂質とタンパク質のアミノ基の数はほぼ同数

O

O

O

O

HOPO

O

OHH2N

680 -Amino residue/ LDL particle

J. P. Segrest et al. / J. Lipid. Res 42 (2001)

Glycation rate0-20%

136 -Amino residue/ LDL particle

137 Amino residue/ LDL particle

Glycationを受けるPEとApoBのアミノ基の数Glycationを受けるPEとApoBのアミノ基の数

Means ± S.D. (n=3), P<0.05

Phosphatidylethanolamine＊

Total cholesterol, Free cholesterol, Triglyceride Phosphatidylcholine, Sphingomyelin, LysophosphatidylcholineLipid hydroperoxides

Not changed

Time (h)

ApoB protein

0

25

50

75

100

0 24 48 72

mol

%

Time (h)

(+)Glu

(-)Glu

mol

%

0

25

50

75

100

0 24 48 72

＊

(+)Glu

(-)Glu

グリケーションによりアミノリン脂質はタンパク質より速く減少した

アミノリン脂質以外の脂質はグリケーションの影響を受けなかった

Changes of Lipid and Protein in Human LDLs by Maillard Reaction Changes of Lipid and Protein in Human LDLs by Maillard Reaction

0

1

2

3

4

0 24 48 72

Lipids

Proteins

dpm

(x1

04 )

Means ± S.D. (n=3), *P < 0.05

＊

＊

ラベル化されたGlucoseが、脂質画分に多く移⾏した

Glucoseはタンパク質よりアミノリン脂質との反応性が高い

PE Amadori-PE LDL

PC

SM

PEAmadori-PE

Silica gel plateCHCl3:MeOH:CH3COOH:H2O = 30:10:4:2 (v/v/v/v)

Time (h)

Radioactive Comparison between Lipid and Protein GlycationsRadioactive Comparison between Lipid and Protein Glycations

HPLC-ELSD conditionsColumn :TOSOH TSK-GEL ODS-80Ts 4.6 x 150 mmMobile phase :MeOH:0.5 M Ammonium acetate = 99:1 (v/v)Flow rate :1.0 mL/min Column oven :35 oCELSD :gas pressure 2.0 bar, drift tube temp 60oC

Retention time (min)

ELSD

resp

onse

160 4 8 12

Amadori-PE

Dioleoyl-PE

Schiff-PE

光散乱検出器ELSD

糖化脂質は光散乱検出器により定量可能であった

検出限界0.5 ng

Quantitative Analysis of Glycated LipidsQuantitative Analysis of Glycated Lipids

Glucose Concentration Reaction Temperature

0

0.40

0.80

0.12

0.16

0.20

0.24

0.28

0 100 200 300 400 500Glucose (mM)

(mM)

Dioleoyl-PE

Schiff-PE

Temp (oC)35 40 45 50 55 60

0

0.40

0.80

0.12

0.16

0.20

0.24

0.28

Schiff-PE

Dioleoyl-PE

(mM)

Means ± S.D. (n=3)

脂質グリケーションはグルコース濃度依存的であり反応温度は影響しなかった

37℃, 2 h 2 h

Effects of Glucose and Temperature in GlycationEffects of Glucose and Temperature in Glycation

脂質グリケーションはメタノール濃度に依存的pHの影響を強く受けた

4 5 6 7 8 9 10pH

50 60 70 80MeOH in phosphate buffer (%)

0

0.40

0.80

0.12

0.16

0.20

0.24

0.40

0.80

0.12

0.16

0.20

0.24

0

Schiff-PE

Dioleoyl-PE

(mM) (mM)

Schiff-PE

MeOH Concentration pH

Phosphatebuffer

Tris-HCl buffer

Means ± S.D. (n=3)

37℃, 2 h 37℃, 2 h

Effects of MeOH Concentration and pHEffects of MeOH Concentration and pH

PEから糖化脂質が効率よく⽣成する反応系が確⽴できた

糖化脂質の定量HPLC-ELSD （光散乱検出器）

PE (0.3 mM)Glucose （500 mM）

70% MeOH/phosphate buffer （pH 7.4）（37℃, 2-48 h）

Test for Inhibitors of Lipid GlycationTest for Inhibitors of Lipid Glycation

Schiff-PEの⽣成を阻害すればAmadori-PEの⽣成が阻害できる

(mM)

12 24 36 48Time (h)

PE

Schiff-PE

Amadori-PE

mean ± S.D, n=3

0

0.04

0.12

0.20

0.28

0

0.08

0.16

0.24

Schiff-PEの⽣成が最⼤Amadori-PEの⽣成

2

Generation of Glycated Lipids (Amadori-PE)Generation of Glycated Lipids (Amadori-PE)

0 20 40 60 80 100

RutinQuercetin

Pyridoxal 5'-phosphatePyridoxal

PyridoxaminePyridoxineCarnosine

Aminoguanidine

L-Cysteine L-Lysine Control

Schiff-PE (% of control)

**

Ascorbic acid-Tocopherol *

Final conc. 1 mM37oC, 2 h

mean ± S.D, n=3, *P<0.05

ピリドキサール類はSchiff-PEの⽣成を阻害した

ResultResult

PEとピリドキサール類の反応により脂質グリケーションが競争的に阻害された

PE

＋O

OHHO

OHHO

HOR1O

O

R2O

O

HOPO

O

OHH2N

0.02 %

OHOHO

OHOH

OH

99.8 %

R1O

O

R2O

O

HO

PO

O

OHN

OH

OHOH

OH

HOR1O

O

R2O

O

HOPO

O

OH

HNO

OH

OHHO

OHSchiff-PE Amadori-PE

Glycation between PE and Glucose

D-Glucose

脂質とグルコースのグリケーション反応はグルコースの開環率に制約される

PL-PE

R1O

O

R2O

O

HOPO

O

OHN

HO

NOH

Pyridoxal (PL)

OHN

HO O

PE

R1O

O

R2O

O

HOPO

O

OHH2N＋

PLP-PE

R1O

O

R2O

O

HOPO

O

OHN

H2O3PO

NOH

Pyridoxal 5’-phosphate (PLP)

OPO3H2N

HO O

PE

R1O

O

R2O

O

HOPO

O

OHH2N＋

脂質とピリドキサール類の反応は制約を受けない

Reaction between PE and Pyridoxal

Reaction of PE with Pyridoxal or GlucoseReaction of PE with Pyridoxal or Glucose

アルデヒド基を有する物質は脂質グリケーションの阻害に有効か？

グリオキサール類やグリセルアルデヒドは、脂質グリケーションを阻害しなかった

OO

OH

HO OO

O

Glyoxal(GO)

Methyl glyoxal(MGO)

DL-Glyceraldehyde(Gly)

C GO MGO Gly0

20

40

60

80

100Sc

hiff-

PE(%

of c

ontr

ol)

mean ± S.D, n=3, *P<0.05

1 mM37 oC, 2 h

Glycation Inhibitory Effect of AldehydesGlycation Inhibitory Effect of Aldehydes

アミノリン脂質とピリドキサール類の付加体がヒト赤血球中に存在する

m/z900 950 10251000925 1050 1075 1100

100

80

60

40

20

0

Rel

ativ

e In

tens

ity (%

)

975

16:0-18:1(Diacyl)

16:0-18:2(Diacyl)

16:0-20:4(Diacyl)

18:0-18:218:1-18:1(Diacyl)

18:1-18:2(Diacyl)

16:0-16:0(Alkyl)

18:1-20:4(Diacyl)

18:0-20:4(Diacyl)

16:0-22:618:2-20:4(Diacyl)

18:1-22:6(Diacyl)

18:0-22:6(Diacyl)

1023.8

16:0-20:4(Plasma)16:0-18:1

(Plasma)16:0-18:2

(Alkyl)

16:0-18:0(Plasma)16:0-18:1

(Alkyl)

18:1-20:4(Plasma)16:0-22:6

(Alkyl)

18:0-20:416:0-22:4(Plasma)18:1-22:4

(Alkyl)

18:0-22:6(Plasma)18:1-22:6

(Alkyl)

O

O

O

O

HOPO

O

OH

HN

O

NOH

PHOO

OH

PE-ピリドキサール5'-リン酸 （16:0-18:1）

Human RBC packed cell (1 mL)

Lipid extraction

LC/MS/MS (Flow injection)

Human RBC preparations

Reduction (NaBH4)

NL (Neutral Loss) Chromatogram of PE-pyridoxal-5‘-phosphate adduct in Human Red Blood CellsNL (Neutral Loss) Chromatogram of PE-pyridoxal-5‘-phosphate adduct in Human Red Blood Cells

lipoprotein lipase activityの低下

膵細胞の破壊

Generation of active oxygen species

Poor Insulin Secretion

Increase in VLDL

High blood glucose level

Hyperlipidemia

STZN-(Methylnitrosocarbamoyl)-

α-D-glucosamine

N. Rakieten., et al. / Cancer. Chemother. Rep. 29 (1963)STZ誘導型糖尿病モデルラットは1型糖尿病モデル

STZ (Streptozotocin) Induced Diabetic Model RatsSTZ (Streptozotocin) Induced Diabetic Model Rats

4 wk 5 wk 10 wk

解剖PLP投与開始

STZ腹腔内投与70 mg/kg

重症

対照群(C) （n=6）

対照(C)＋PLP (2 mM) 投与群 （n=6）

STZ群 （n=9）

STZ＋PLP (1 mM) 投与群 （n=9）

Male SD rats4 wk

6 wk

＜投与方法＞ピリドキサール5'-リン酸水

溶液を経口自由摂取

測定項目●空腹時血糖値●糖化ヘモグロビン●中性脂質（トリグリセリド、

コレステロール）●リン脂質（PC, PE, SM, LPC）●血漿TBARS●血漿PCOOH●糖化脂質

MethodMethod

0

10

20

30

40

C C+PLP STZ STZ+PLP0

5

10

15

20

C C+PLP STZ STZ+PLPmean ± S.D, n=6-9,

P<0.05 vs C (Control grooup)mean ± S.D, n=6-9

P<0.05 vs C (Control group)

Blo

od g

luco

se (m

M)

Gly

cate

d he

mog

lobi

n (%

)

** *

*

ピリドキサール5'-リン酸は、血漿グルコース濃度糖化ヘモグロビン値を低下させなかった

Blood glucose level Glycated hemoglobin

Blood Glucose Level and Glycated HemoglobinBlood Glucose Level and Glycated Hemoglobin

ピリドキサール5'-リン酸は、血漿脂質組成にほとんど影響を与えない

ParametersControl

(n=6)Control + PLP

(n=6)STZ + PLP

(n=9)STZ(n=9)

Table Phospholipids (PC, PE, SM, LPC), neutral lipids (TG, T-CHO) in plasma.

C, indicates control; PLP, pyridoxal 5'-phosphate; PC, phosphatidylcholine; PE, phosphatidylethanolamine; SM, sphingomyelin; LPC, lysophosphatidylcholine; TG, triglyceride; T-CHO, total cholesterol. Data are expressed as mean ± SD. *P<0.05 vs control groups.

SM 28.8 162.1± 164.8 41.0± 74.3226.7 ± 202.7 90.8±

LPC 51.6236.9± 268.7 53.9± 69.0195.1 ± 218.8± 84.0

158.3 893.0 ± 927.5 175.7± 1815.7 718.0± 1516.5 734.0±PC *10.2 65.6 ± 68.1 15.1± 75.2229.3 ± 205.3 98.4±PE * *

T-CHO ±37.2 15.3 30.9 12.0± 198.1 84.6± 170.6 68.0±* *±89.9 32.5 102.2 9.2± 475.5 158.3± 390.5 176.2±TG * *

Phospholipids (nM)

Neutral lipids (mg/dl)

ParametersControl

(n=6)Control + PLP

(n=6)STZ + PLP

(n=9)STZ(n=9)

Table Phospholipids (PC, PE, SM, LPC), neutral lipids (TG, T-CHO) in plasma.

C, indicates control; PLP, pyridoxal 5'-phosphate; PC, phosphatidylcholine; PE, phosphatidylethanolamine; SM, sphingomyelin; LPC, lysophosphatidylcholine; TG, triglyceride; T-CHO, total cholesterol. Data are expressed as mean ± SD. *P<0.05 vs control groups.

SM 28.8 162.1± 164.8 41.0± 74.3226.7 ± 202.7 90.8±SM 28.8 162.1± 164.8 41.0± 74.3226.7 ± 202.7 90.8±

LPC 51.6236.9± 268.7 53.9± 69.0195.1 ± 218.8± 84.0LPC 51.6236.9± 268.7 53.9± 69.0195.1 ± 218.8± 84.0

158.3 893.0 ± 927.5 175.7± 1815.7 718.0± 1516.5 734.0±PC *158.3 893.0 ± 927.5 175.7± 1815.7 718.0± 1516.5 734.0±PC *10.2 65.6 ± 68.1 15.1± 75.2229.3 ± 205.3 98.4±PE * *10.2 65.6 ± 68.1 15.1± 75.2229.3 ± 205.3 98.4±PE * *

T-CHO ±37.2 15.3 30.9 12.0± 198.1 84.6± 170.6 68.0±* *T-CHO ±37.2 15.3 30.9 12.0± 198.1 84.6± 170.6 68.0±* *±89.9 32.5 102.2 9.2± 475.5 158.3± 390.5 176.2±TG * *±89.9 32.5 102.2 9.2± 475.5 158.3± 390.5 176.2±TG * *

Phospholipids (nM)

Neutral lipids (mg/dl)

Changes in Plasma LipidsChanges in Plasma Lipids

0

2

4

6

8

10

C C+PLP STZ STZ+PLP

Plas

ma

TBAR

S (

M)

* **

mean ± S.D, n=6-9, *P<0.05 vs S(STZ group)

ピリドキサール5'-リン酸の摂取は高血糖状態での脂質過酸化を抑制する

mean ± S.D, n=6-9, *P<0.05 vs S (STZ group)

0

20

40

60

80

100

120

C C+PLP STZ STZ+PLPPl

asm

a PC

OO

H (p

mol

/mL

plas

ma)

* *

*

Plasma TBARS Peroxided Lipids (PCOOH）

TBARS and Peroxided Lipids (PCOOH) in PlasmaTBARS and Peroxided Lipids (PCOOH) in Plasma

0

200

400

600

800

1000

1200

1400

16:0-18:2

Amad

ori-P

E (p

mol

/mL

plas

ma)

C C+P

LPST

ZST

Z +

PLP

mean ± S.D, n=6-9, *P<0.05 vs C (Control group)

**

** *

*

*

*

Amadori-PE0

0.3

0.5

0.8

1.0

1.3

1.5

1.8

C C+PLP STZ STZ+PLP

Amad

ori-P

E/PE

(mol

%)

mean ± S.D, n=6-9, *P<0.05 vs S (STZ group)

*

*

*** *

ピリドキサール5'-リン酸の摂取は高血糖状態での脂質グリケーションを阻害する

16:0-18:1 18:2-20:4 18:0-22:6

Glycated Lipids in Rat Plasma (Amadori-PE)Glycated Lipids in Rat Plasma (Amadori-PE)