げっ歯類における数種の異なる疾病モデルを用いたグリーンプロポリスの防御効果に関する研究

Title

げっ歯類における数種の異なる疾病モデルを用いたグリー

_{ンプロポリスの防御効果に関する研究( 本文(Fulltext) )}

Author(s)

土屋, 祐弥

Report No.(Doctoral

Degree)

博士(獣医学) 甲第510号

Issue Date

2018-09-21

Type

博士論文

Version

ETD

URL

http://hdl.handle.net/20.500.12099/77271

※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。





ࡆࡗṑ㢮࡟࠾ࡅࡿᩘ✀ࡢ␗࡞ࡿ⑌⑓ࣔࢹࣝࢆ⏝࠸ࡓ

ࢢ࣮ࣜࣥࣉ࣏ࣟࣜࢫࡢ㜵ᚚຠᯝ࡟㛵ࡍࡿ◊✲









 ᖺ



ᒱ㜧኱Ꮫ኱Ꮫ㝔㐃ྜ⋇་Ꮫ◊✲⛉

㸦ᒱ㜧኱Ꮫ㸧





ᅵᒇ ♸ᘺ

1

ࡆࡗṑ㢮࡟࠾ࡅࡿᩘ✀ࡢ␗࡞ࡿ⑌⑓ࣔࢹࣝࢆ⏝࠸ࡓ

ࢢ࣮ࣜࣥࣉ࣏ࣟࣜࢫࡢ㜵ᚚຠᯝ࡟㛵ࡍࡿ◊✲

ᅵᒇ ♸ᘺ

2

┠

┠ḟ

␎ㄒ୍ぴ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 4 ᗎㄽ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 6 ➨1❶ EEBGP ࡢ⸆≀ㄏⓎᛶᛴᛶ⫢㞀ᐖࣔࢹࣝ࡟ᑐࡍࡿ㜵ᚚຠᯝࡢ᳨ウ ⥴ゝ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 12 ᮦᩱ࠾ࡼࡧ᪉ἲ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 14 ⤖ᯝ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 19 ⪃ᐹ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 22 ᑠᣓ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 27 ᅗ⾲ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 29 ➨2❶ EEBGP ࡢ AA ࢔࣑ࣟ࢖ࢻ⑕ࣔࢹࣝ࡟ᑐࡍࡿ㜵ᚚຠᯝࡢ᳨ウ ⥴ゝ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 43 ᮦᩱ࠾ࡼࡧ᪉ἲ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 44 ⤖ᯝ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 45 ⪃ᐹ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 46 ᑠᣓ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 48 ᅗ⾲ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 49 ➨3❶ EEBGP ࡢ HEV-9 ឤᰁ࡟ࡼࡿᛴᛶ⬻⅖ࣔࢹࣝ࡟ᑐࡍࡿ㜵ᚚຠᯝࡢ᳨ウ ⥴ゝ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 57 ᮦᩱ࠾ࡼࡧ᪉ἲ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 57 ⤖ᯝ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 54 ⪃ᐹ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 60 ᑠᣓ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 61 ᅗ⾲ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 62

3

⥲ྜウㄽ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 65

⤖ㄽ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 69

ㅰ㎡ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 70

4

␎

␎ㄒ୍ぴ

AA: Amyloid A ACTB: β-Actin

ALT: Alanine aminotransferase APAP: N-Acethyl-p-aminophenol AST: Aspartate aminotransferase CAPE: Caffeic acid phenethyl ester CAT: Catalase

CCL: CC-motif chemokine CYP2E1: Cytochrome P450 2E1 CXCL: CXC-motif chemokine

DPPH: 2, 2-Diphenyl-1-picrylhydrazyl

EEBGP: Ethanol extract of Brazilian green propolis EHV-9: Equine herpesvirus-9

GAPDH: Glyceraldehyde-3-phosphate dehydrogenase GSTM1: Glutathione-S-transferase M1 H.E.: Hematoxylin-Eosin IL: Interleukin LPS: Lipopolysaccharide NAPQI: N-Acethylbenzoquinoneimine NF-κB: Nuclear factor-κB

NOS2: Nitric oxide synthase-2

NRF2: Nuclear-factor erythroid-derived 2-related factor-2 SAA: Serum amyloid A

5 GPX: Grutathione peroxidase

GSR: Glutathione reductase

PPARα: Peroxisome proliferator-activated receptor α SULT1A1: Sulfotransferase 1A1

TNF-α: Tumor necrosis factor-α

6

ᗎ

ᗎㄽ

 ᡃࡀᅜࡢࠕᖹᆒᑑ࿨ࠖࡣ, 2013 ᖺ᫬Ⅼ࡛⏨ᛶࡀ 80.21 ṓ, ዪᛶࡀ 86.61 ṓ࡛࠶ࡿ୍᪉, ௓ㆤࢆཷࡅࡓࡾ ᐷࡓࡁࡾ࡟࡞ࡗࡓࡾࡏࡎ᪥ᖖ⏕άࢆ㏦ࢀࡿᮇ㛫ࢆ♧ࡍࠕ೺ᗣᑑ࿨ࠖࡀ, ⏨ᛶࡀ 71.19 ṓ, ዪᛶࡀ 74.21 ṓ ࡜, ୧⪅ࡢ㛫࡟ 10 ṓ๓ᚋࡢᕪࡀ࠶ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[1]ࠋ୍᪉, ᅜࡢ་⒪㈝ࡣᖺࠎቑຍࡢ୍㏵ࢆࡓ ࡝ࡗ࡚࠾ࡾ, 2015 ᖺ࡟ࡣ 42 ඙෇࡟ࡶ㐩ࡋ࡚࠸ࡿࡀ, ࡑࡢ⣙ 60%ࡣ 65 ṓ௨ୖࡢ㧗㱋⪅ࢆᑐ㇟࡜ࡋࡓࡶࡢ ࡛࠶ࡾ㸦ᅗ1-1㸧[2], ᖹᆒᑑ࿨࡜೺ᗣᑑ࿨࡜ࡢ஋㞳ࡀ኱ࡁ࡞ၥ㢟࡜࡞ࡗ࡚࠸ࡿࠋࡑࡢࡓࡵ, ᪥ᮏᨻᗓࡣᕷ ㈍⏝་⸆ရࢆ⏝࠸ࡓࢭࣝࣇ࣓ࢹ࢕ࢣ࣮ࢩࣙࣥ࡟ࡼࡿ་⒪㈝ࡢᢚไࢆ㐍ࡵࡿ࡜࡜ࡶ࡟[3], ⱝᖺᒙࡸ୰㧗ᖺ ᒙࢆྵࡴẚ㍑ⓗⱝ࠸ୡ௦࠿ࡽࡢ⑌⑓ࡢண㜵ࢆ᥎㐍ࡋ, ≉࡟㣗ရࡢ᭷ࡍࡿ೺ᗣቑ㐍ຠᯝ࡟╔┠ࡋ࡚࠸ࡿࠋ 㣗ရࡣ, ೺ᗣ㣗ရ㸦ࢧࣉ࣓ࣜࣥࢺ㸧ࢆྵࡴ୍⯡㣗ရ࡜ಖ೺ᶵ⬟㣗ရࡢ 2 ࡘ࡟኱ࡁࡃศ㢮ࡉࢀ, ࡉࡽ࡟ಖ ೺ᶵ⬟㣗ရࡣ≉ᐃಖ೺⏝㣗ရ, ᰤ㣴ᶵ⬟㣗ရ, ᶵ⬟ᛶ⾲♧㣗ရ࡟ศࡅࡽࢀ, ࡑࢀࡒࢀ 1991 ᖺ, 2001 ᖺ, 2015 ᖺࡼࡾ㐠⏝ࡀ㛤ጞࡉࢀ࡚࠸ࡿ㸦ᅗ 1-2㸧[4, 5]ࠋࡇࢀࡽಖ೺ᶵ⬟㣗ရ࡛ࡣ, ୍⯡㣗ရ࡜ࡣ␗࡞ࡾ, ೺ ᗣቑ㐍࡟㛵ࢃࡿᶵ⬟ᛶ⾲♧ࡀㄆࡵࡽࢀ࡚࠾ࡾ, 2018 ᖺ 3 ᭶ࡢ᫬Ⅼ࡛, ≉ᐃಖ೺⏝㣗ရࡢチྍᩘࡣ 1, 000 ௳ࢆ[6], ᶵ⬟ᛶ⾲♧㣗ရࡢⓏ㘓ᩘࡣ 1, 300 ௳ࢆ㉸࠼࡚࠾ࡾ[7], ௒ᚋࡉࡽ࡟ቑຍࡍࡿࡇ࡜ࡀண᝿ࡉࢀࡿࠋ ࡇࢀࡽ㣗ရࡢ⏝㏵ᣑ኱࡟క࠸, ࢧࣉ࣓ࣜࣥࢺࡢ೺ᗣቑ㐍ຠᯝ࡟㛵ࡍࡿ࢚ࣅࢹࣥࢫࢆྲྀᚓࡍࡿࡇ࡜ࡢ㔜せ ᛶࡀ㧗ࡲࡗ࡚࠸ࡿࠋ  ࣉ࣏ࣟࣜࢫࡣ, ࣑ࢶࣂࢳࡀᶞᮌࡢ᪂ⱆࡸᶞᾮ࡜⮬ࡽࡢศἪ≀ࢆΰྜࡋ࡚సࡿᶞ⬡≧≀㉁࡛࠶ࡾ, ఏ⤫ ་⸆ࡸࢧࣉ࣓ࣜࣥࢺ⣲ᮦ࡜ࡋ࡚฼⏝ࡉࢀ࡚࠸ࡿ[8]ࠋࣉ࣏ࣟࣜࢫࡢྵ᭷ᡂศࡣ, ᮦᩱ࡜࡞ࡿ᳜≀ࡸᢳฟ᮲ ௳࡟ࡼࡾ␗࡞ࡿࠋ࣮ࣚࣟࢵࣃࡸ୰ᅜ࡛ࡣ, ࣏ࣉࣛ࡟⏤᮶ࡍࡿࣉ࣏ࣟࣜࢫࡢ࢚ࢱࣀ࣮ࣝᢳฟ≀ࡀᗈࡃ฼⏝ࡉ ࢀ࡚࠾ࡾ, ࢡࣜࢩࣥࡸࣆࣀࢭࣥࣈࣜࣥ, ࢣࣝࢭࢳࣥ, ࢞ࣛࣥࢠࣥ➼ࡢࣇࣛ࣎ࣀ࢖ࢻࡢ࡯࠿, ≉ᚩⓗ࡞ᡂศ ࡜ࡋ࡚࢝ࣇ࢙㓟ࣇ࢙ࢿࢳ࢚ࣝࢫࢸࣝ㸦CAPE㸧ࢆྵࡴ㸦⾲ 1-1㸧[9-11]ࠋ࣏ࣉࣛࢱ࢖ࣉࡢࣉ࣏ࣟࣜࢫ࡛ࡣ, ᢠ࢘࢖ࣝࢫస⏝ࡸᢠࡀࢇస⏝, ᢠ㓟໬స⏝, ᢠ⅖⑕స⏝࡞࡝ࡀ▱ࡽࢀ࡚࠾ࡾ[9], ࣐࢘ࢫࡸࣛࢵࢺ࡟࠾࠸ ࡚⹫⾑෌℺ὶ࡟క࠺ᚰẘᛶ[12], ࢻ࢟ࢯࣝࣅࢩࣥࡸࢩࢫࣉࣛࢳࣥ࡟ࡼࡿ⭈ẘᛶ[13], ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥ 㸦N-࢔ࢭࢳࣝ-p-࢔࣑ࣀࣇ࢙ࣀ࣮ࣝ; APAP㸧ࡸᅄሷ໬Ⅳ⣲࡟ࡼࡿ⫢ẘᛶ[14, 15]ࢆᢚไࡍࡿࡇ࡜ࡀሗ࿌ࡉ ࢀ࡚࠸ࡿࠋ୍᪉, ᪥ᮏ࡛ࡣ୺࡟ࣈࣛࢪࣝ⏘ࡢࣉ࣏ࣟࣜࢫࡀࢧࣉ࣓ࣜࣥࢺ࡜ࡋ࡚฼⏝ࡉࢀ࡚࠸ࡿࠋ⃰࠸⥳Ⰽ ࡀ≉ᚩⓗ࡞ࣈࣛࢪࣝ⏘ࢢ࣮ࣜࣥࣉ࣏ࣟࣜࢫ㸦Brazilian green Propolis; BGP㸧ࡣ, ୺࡟࢟ࢡ⛉ࡢBaccharis

7 dracunculifolia D.C.ࢆᮦᩱ࡜ࡋ࡚సࡽࢀ㸦࢔ࣞࢡࣜࣥࢱ࢖ࣉ㸧, ࡑࡢ࢚ࢱࣀ࣮ࣝᢳฟ≀㸦Ethanolic extract of BGP; EEBGP㸧ࡣ, ࢔ࣝࢸࣆࣜࣥ C ࡸࣂࢵ࢝ࣜࣥ, ࢻࣝࣃࢽࣥ➼ࡢࣉࣞࢽࣝ໬ࣇ࢙ࢽࣝࣉࣟࣃ ࣀ࢖ࢻ㢮ࡸ, ࢣࣥࣇ࢙࣮ࣟࣝࡸࢣࣥࣇ࢙ࣛ࢖ࢻ➼ࡢࣇࣛ࣎ࣀ࢖ࢻ㢮ࢆྵࡳ㸦⾲ 1-1, ᅗ 1-3㸧[11], ᢠ⳦స ⏝, ᢠ࢘࢖ࣝࢫస⏝, ᢠ㓟໬స⏝, ᢠ⅖⑕స⏝࡞࡝ࡀ▱ࡽࢀ࡚࠸ࡿ[16, 17]ࠋEEBGP ࡣ, ື≀ᐇ㦂࡟࡚⢾ ᒀ⑓ᛶ⭈⑕࡟ᑐࡋ࡚ಖㆤస⏝ࢆ♧ࡋ[18], α-ࢼࣇࢳࣝ࢖ࢯࢳ࢜ࢩ࢔ࢿ࣮ࢺ[19], D-࢞ࣛࢡࢺࢧ࣑ࣥ[20]࠾ࡼ ࡧỈᾐᣊ᮰ࢫࢺࣞࢫ[21]࡟ࡼࡿ⾑୰⫢㞀ᐖ࣐࣮࣮࢝ࡢቑຍࢆᢚไࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ, ࡑࡢ࣓࢝ࢽࢬ࣒ ࡜ࡋ࡚EEBGP ࡢ᭷ࡍࡿᢠ㓟໬స⏝ࡀ⪃࠼ࡽࢀ࡚࠸ࡿࡀ[18, 19], ⤌⧊࡛ࣞ࣋ࣝࡢኚ໬ࢆホ౯ࡋࡓሗ࿌ࡣ ࡯࡜ࢇ࡝࡞࠸ࠋ ᮏ◊✲࡛ࡣ, ␗࡞ࡿ࣓࢝ࢽࢬ࣒࡟ᇶ࡙࠸࡚⑌⑓ࢆⓎ⑕ࡍࡿ 4 ✀ࡢື≀ࣔࢹࣝ࡟࠾࠸࡚, EEBGP ࡀ㜵ᚚ ຠᯝࢆ♧ࡍ࠿ྰ࠿᳨ウࡋ, ᢠ㓟໬స⏝ࡸ⅖⑕ᢚไస⏝࡜ࡢ㛵ಀᛶࢆ☜ㄆࡋࡓࠋ➨ 1 ❶࡛ࡣ, 2 ✀ࡢ⫢ẘᛶ ≀㉁㸦APAP ࠾ࡼࡧᅄሷ໬Ⅳ⣲㸧࡟ࡘ࠸࡚, ᢞ୚᮲௳ࢆỴᐃࡍࡿࡓࡵ, ࡇࢀࡽࡢ≀㉁ࡢ௦ㅰ࡟㛵ࢃࡿ⸆≀ ௦ㅰ㓝⣲ࡢ㑇ఏᏊⓎ⌧࡜ࢢࣝࢱࢳ࢜ࣥ㔞࡟ᑐࡋ࡚ᦤ㣵᮲௳࠾ࡼࡧᢞ୚ࡢࢱ࢖࣑ࣥࢢࡢ㐪࠸ࡀ୚࠼ࡿᙳ㡪 ࡟ࡘ࠸࡚ㄪ࡭, ⫢⮚ࡢയᐖࡢ᭷↓࠾ࡼࡧࡑࡢ⛬ᗘ࡜ࡢ㛵㐃࡟ࡘ࠸᳨࡚ウࡋࡓࠋ⥆࠸࡚, APAP ࠶ࡿ࠸ࡣᅄ ሷ໬Ⅳ⣲࡟ࡼࡿ⫢⣽⬊ቯṚ࡟ᑐࡍࡿ EEBGP ࡢ㜵ᚚຠᯝࢆ᳨ウࡍࡿ࡜࡜ࡶ࡟, ᢠ㓟໬㓝⣲࠾ࡼࡧ⅖⑕㛵 㐃ᅉᏊࡢ㑇ఏᏊⓎ⌧࡟ᑐࡍࡿᙳ㡪ࢆ☜ㄆࡋ, 㜵ᚚຠᯝ࡜ࡢ㛵㐃ࢆㄪ࡭ࡓࠋ➨ 2 ❶࡛ࡣᛴᛶ⫢㞀ᐖࣔࢹࣝ࡜ ࡣ␗࡞ࡿ㞀ᐖࣔࢹࣝ࡜ࡋ࡚, ࢔࣑ࣟ࢖ࢻ A㸦AA㸧࢔࣑ࣟ࢖ࢻ⑕࡟ᑐࡍࡿ EEBGP ࡢ㜵ᚚຠᯝࢆ᳨ウࡍࡿ ࡜࡜ࡶ࡟, ᢠ㓟໬㓝⣲࠾ࡼࡧ⅖⑕㛵㐃ᅉᏊࡢ㑇ఏᏊⓎ⌧࡟ᑐࡍࡿᙳ㡪ࢆㄪ࡭, 㓟໬ࢫࢺࣞࢫ࠾ࡼࡧ⅖⑕ ཯ᛂ࡜ࡢ㛵㐃࡟ࡘ࠸࡚☜ㄆࡋࡓࠋࡲࡓ, EEBGP ࡟ࡣᢠ࢘࢖ࣝࢫస⏝ࡢሗ࿌ࡀ࠶ࡿࡓࡵ[16, 17], ➨ 3 ❶࡛ ࡣ, ࣐࢘࣊ࣝ࣌ࢫ࢘࢖ࣝࢫ㸦EHV㸧-9 ឤᰁ࡟క࠺ᛴᛶ⬻⅖࡟ᑐࡍࡿ EEBGP ࡢ㜵ᚚຠᯝࢆ᳨ウࡋ, ేࡏ ࡚⅖⑕㛵㐃ᅉᏊࡢ㑇ఏᏊⓎ⌧࡟୚࠼ࡿᙳ㡪ࢆ᳨ウࡋࡓࠋ

8

⾲ 1-1. ࢔ࣞࢡࣜࣥ㸦Alecrim㸧ࢱ࢖ࣉࣉ࣏ࣟࣜࢫ࡜࣏ࣉࣛ㸦Poplar㸧ࢱ࢖ࣉࣉ࣏ࣟࣜࢫࡢྵ᭷ᡂศࡢ㐪 ࠸㸦Zhang J et al., 2016 [11]ࡼࡾᢤ⢋୍࣭㒊ᨵኚ㸧

-: ᮍ᳨ฟࡲࡓࡣᐃ㔞㝈⏺௨ୗ

⾲୰ࡢ໬Ꮫ≀㉁๓ࡢᩘᏐࡣᅗ1-3 ࡟ᑐᛂࡍࡿࠋ

Alecrim type Poplar type Phenylpropanoids (Cinnamic acid derivatives)

䐟 p -Coumaric acid 18.01 2.70

Caffeic acid 1.47 13.35

䐠 Chlorogenic acid 8.98

-Caffeic acid phenetyl ester (CAPE) - 24.43 Prenylated phenylpropanoids 䐡 Artepillin C 107.70 Flavonoids Quercetin - 5.90 䐢 Kaempferol 0.99 1.85 Galangin - 22.23 䐣 Kaempferide 11.18 -Pinocembrin 1.13 44.55 Chrysin - 66.89 Chemicals Contents (mg/g)

9

ᅗ1-1. ᅜẸ་⒪㈝ࡢ᥎⛣㸦ᖹᡂ 27 ᖺᗘ ᅜẸ་⒪㈝ࡢᴫἣ[2]ࡼࡾᢤ⢋㸧

10

ᅗ1-2. ୍⯡㣗ရ, ಖ೺ᶵ⬟㣗ရ, ་⸆ရࡢศ㢮㸦ᶵ⬟ᛶ⾲♧㣗ရࡗ࡚ఱ?[5]ࡼࡾᢤ⢋㸧

 ࢧࣉ࣓ࣜࣥࢺࢆྵࡴ୍⯡㣗ရ࡛ࡣ, ຠᯝ࣭ຠ⬟➼ࡢᶵ⬟ᛶ⾲♧ࡣ୍ษㄆࡵࡽࢀ࡚࠸࡞࠸ࡀ, ಖ೺ᶵ⬟㣗ရ࡛ࡣ, ࣄࢺ⮫ᗋ ヨ㦂ࡢ⤖ᯝ࡟ᇶ࡙ࡃࡶࡢ࡟㝈ࡾᶵ⬟ᛶ⾲♧ࡀㄆࡵࡽࢀ࡚࠸ࡿࠋ

11

ᅗ1-3. ࣈࣛࢪࣝ⏘ࢢ࣮ࣜࣥࣉ࣏ࣟࣜࢫࡢ࢚ࢱࣀ࣮ࣝᢳฟ≀㸦EEBGP㸧୰ࡢ୺࡞ྵ᭷ᡂศ

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➨

➨

1 ❶

13 ࠙⥴ゝࠚ ୍⯡ⓗ࡞ゎ⇕㙠③⸆࡛࠶ࡿ APAP ࡣ, ࣄࢺ࡟࠾࠸࡚㐣๫㔞ࡢ᭹⏝࡟ࡼࡗ࡚⫢㞀ᐖࢆ⏕ࡌࡿࡇ࡜ࡀ▱ࡽ ࢀ࡚࠾ࡾ, ື≀࡛ࡶࣄࢺ࡜ྠᵝࡢ࣓࢝ࢽࢬ࣒࡛㞀ᐖࢆ⏕ࡌࡿࡇ࡜࠿ࡽ⸆≀ᛶࡢ⫢㞀ᐖࣔࢹࣝ࡜ࡋ࡚ᗈࡃ ฼⏝ࡉࢀ࡚࠸ࡿ[22]ࠋAPAP ࡟ࡼࡿ⫢ẘᛶⓎ⌧࣓࢝ࢽࢬ࣒ࢆᅗ 2-1 ࡟♧ࡍࠋ⤒ཱྀᦤྲྀࡋࡓ APAP ࡣᑠ⭠࠿ ࡽཷືᣑᩓ࡟ࡼࡾ 90%௨ୖࡀ྾཰ࡉࢀ[23-25], ኱㒊ศࡣࢢࣝࢡࣟࣥ㓟ᢪྜ࠾ࡼࡧ◲㓟ᢪྜࢆཷࡅ࡚ᒀ୰ ࡬᤼ἥࡉࢀࡿࠋ୍㒊ࡣࢩࢺࢡ࣒ࣟP450 2E1㸦CYP2E1㸧࡟ࡼࡾ௦ㅰࡉࢀ࡚཯ᛂᛶࡢ㧗࠸N-࢔ࢭࢳࣝ࣋ ࣥࢰ࢟ࣀࣥ࢖࣑ࣥ㸦NAPQI㸧࡜࡞ࡿࡀ, NAPQI ࡣ㏿ࡸ࠿࡟ࢢࣝࢱࢳ࢜ࣥᢪྜࢆཷࡅ࡚ᒀ୰࡬࡜᤼ἥࡉࢀ ࡿ[23, 25, 26]ࠋࡋ࠿ࡋ࡞ࡀࡽ, ୍ᗘ࡟኱㔞ࡢ APAP ࢆᦤྲྀࡍࡿ࡜, ࢢࣝࢡࣟࣥ㓟ᢪྜ࠾ࡼࡧ◲㓟ᢪྜࡢ㣬 ࿴࡟కࡗ࡚NAPQI ࡢ⏕ᡂ㔞ࡀቑຍࡋ, ᤼ἥࡉࢀ࡞࠿ࡗࡓ NAPQI ࡀ⣽⬊ෆࡢ࣑ࢺࢥࣥࢻࣜ࢔ࢱࣥࣃࢡ࡜ 㠀≉␗ⓗ࡟⤖ྜࡍࡿࠋࡑࡢ⤖ᯝ, ATP ྜᡂࡀ㜼ᐖࡉࢀ࡚⣽⬊ᶵ⬟࡟␗ᖖࢆࡁࡓࡍ࡜࡜ࡶ࡟, 㐣㓟໬཯ᛂ࡟ ࡼࡗ࡚⭷⬡㉁, ࢱࣥࣃࢡ㉁, DNA ࡀᦆയࡋ, ⫢⣽⬊ቯṚࡀ⏕ࡌࡿ[27-29]ࠋᐇ㝿࡟, ᢠ㓟໬㓝⣲࡛࠶ࡿ Mn-ࢫ࣮ࣃ࣮࢜࢟ࢩࢻࢹ࢕ࢫ࣒ࢱ࣮ࢮ㸦Mn-SOD㸧ࡸࢢࣝࢱࢳ࢜ࣥ࣌ࣝ࢜࢟ࢩࢲ࣮ࢮ㸦GPX㸧-1 ࢆḞᦆࡋࡓ ࣐࢘ࢫ[30, 31]ࡸࡇࢀࡽࡢᢠ㓟໬㓝⣲ࡢ㑇ఏᏊⓎ⌧࡟㛵ࢃࡿ nuclear-factor erythroid-derived 2-related factor㸦NRF㸧2 ࢆḞᦆࡋࡓ࣐࢘ࢫ࡛ APAP ࡟ࡼࡿ⫢㞀ᐖࡀᝏ໬ࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[32]ࠋࡲࡓ, 㓟 ໬ࢫࢺࣞࢫ࡟ࡼࡿ⫢⣽⬊ቯṚ࡟ຍ࠼࡚, APAP ᢞ୚࡟ࡼࡿ⫢㞀ᐖ࡟ࡣ⅖⑕཯ᛂࡀ㔜せ࡛࠶ࡿࡇ࡜ࡀ᫂ࡽ࠿ ࡜࡞ࡗ࡚࠸ࡿ[33, 34]ࠋ⫢⮚ᅛ᭷ࡢ࣐ࢡࣟࣇ࢓࣮ࢪ࡛࠶ࡿࢡࢵࣃ࣮⣽⬊ࡣ, ቯṚࡋࡓ⣽⬊࠿ࡽᨺฟࡉࢀࡿ damage-associated molecular patterns ࡟ᛂ⟅ࡋ࡚ࢧ࢖ࢺ࢝࢖ࣥࡸࢣࣔ࢝࢖ࣥࢆ⏘⏕ࡋ, ⅖⑕ᛶ⣽⬊ࢆ ㄏᘬࡍࡿࡇ࡜࡛ቯṚࡋࡓ⣽⬊ࢆ㝖ཤࡍࡿࡀ, 㐣๫࡞⅖⑕཯ᛂࡣṇᖖ࡞⫢⣽⬊ࢆ◚ቯࡋ, ⫢⣽⬊ቯṚࡢ⠊ ᅖࢆᣑ኱ࡉࡏࡿ[33-35]ࠋඛ⾜◊✲࡟ࡼࡾ, ⅖⑕ᛶࢧ࢖ࢺ࢝࢖࡛ࣥ࠶ࡿ Interlukin㸦IL㸧-1 ࡢ⏘⏕ࡸࡑࡢ ࢩࢢࢼࣝఏ㐩[36-38], ࢣࣔ࢝࢖࡛ࣥ࠶ࡿ CC motif chemokine ligand㸦CCL㸧-2 ࡢࢩࢢࢼࣝఏ㐩[39]ࢆ᧠ ஘ࡍࡿ࡜, APAP ࡟ࡼࡿ⫢㞀ᐖࡀ㍍ῶࡉࢀࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗ࡚࠸ࡿࠋ ࣄࢺ࡟࠾ࡅࡿAPAP ࡢ୰ẘ㔞ࡣ 1 ᪥᭱኱᭹⏝㔞㸦4 g/᪥㸧ࡢ 2 ಸ࡟┦ᙜࡍࡿ 8 g/᪥࡜ࡢሗ࿌ࡀ࠶ࡾ, ࡑ ࡢ኱㒊ศࡣ⮬ẅ┠ⓗࡢពᅗⓗ࡞㐣๫㔞ࡢ᭹⏝࡛࠶ࡿ[40]ࠋ୍᪉, ࣄࢺ࡟࠾࠸࡚, ៏ᛶⓗ࡞ከ㔞ࡢ࢔ࣝࢥ࣮ ࣝᦤྲྀୗࡸ, 㛗ᮇࡢ⤯㣗ୗ[40-42]ࡸపᰤ㣴≧ែୗ[40, 41]࡛ࡣ, ୍᪥᭱኱᭹⏝㔞௨ୗࡢᦤྲྀ࡛࠶ࡗ࡚ࡶ⫢ 㞀ᐖࢆ⏕ࡌࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋࡲࡓ, ࣐࢘ࢫ࡛ࡣ, ᢞ୚㔞ࡀྠࡌ࡛࠶ࡿ࡟ࡶ࠿࠿ࢃࡽࡎ, APAP ࡢᢞ ୚᫬้࡟ࡼࡗ࡚Ṛஸ⋡ࡸ⫢⣽⬊ቯṚࡢ⛬ᗘࡀ␗࡞ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[43]ࠋ

14 ࣄࢺ࡟࠾࠸࡚, 㧗⃰ᗘࡢᅄሷ໬Ⅳ⣲⵨Ẽࡢ྾ධࡸ⁐ᾮࡢ᭚㟢ࡣ, ⫢⮚ࡸ⭈⮚, ⚄⤒⣔࡞࡝࡟㞀ᐖࢆ㉳ࡇ ࡍࡀ, ⫢ẘᛶ࡟ࡘ࠸࡚ࡣࣛࢵࢺࡸ࣐࢘ࢫ࡞࡝ࡢື≀࡛ࡶࡼࡃ෌⌧ࡉࢀࡿࡓࡵ, ⫢㞀ᐖࣔࢹࣝ࡜ࡋ࡚ᗈࡃ ฼⏝ࡉࢀ࡚࠸ࡿ[22]ࠋᅄሷ໬Ⅳ⣲࡟ࡼࡿ⫢ẘᛶⓎ⌧࣓࢝ࢽࢬ࣒ࢆᅗ 2-2 ࡟♧ࡍࠋ྾཰ࡉࢀࡓᅄሷ໬Ⅳ⣲ࡣ ⫢⮚࡛CYP2E1 ࡟ࡼࡿ௦ㅰཷࡅ, ࢺࣜࢡ࣓ࣟࣟࢳࣝࣛࢪ࢝ࣝ㸦CCl3࣭㸧ࡸࢺࣜࢡ࣓ࣟࣟࢳࣝ࣌ࣝ࢜࢟ࢩ ࣛࢪ࢝ࣝ㸦CCl3OO࣭㸧ࢆ⏕ᡂࡍࡿ[44-46]ࠋࡇࢀࡽࡢࣛࢪ࢝ࣝࡣ, ⬡㉁⭷ࡢ㓟໬ࡸ⏕య㧗ศᏊ࡬ࡢ㠀≉␗ ⓗ⤖ྜࢆ௓ࡋ࡚⣽⬊ᶵ⬟ࢆ㞀ᐖࡍࡿࡓࡵ, CYP2E1 ࡢⓎ⌧㔞ࡀ≉࡟ከ࠸ᑠⴥ୰ᚰᇦ࡛⫢⣽⬊ቯṚࡀ⏕ࡌ ࡿ[44, 45]ࠋࡲࡓ, ᅄሷ໬Ⅳ⣲ࡣ⫢⮚࡟࠾ࡅࡿ࢔࣏࣏ࣜࢱࣥࣃࢡ㉁ࡢྜᡂࢆ㜼ᐖࡋ, ࢺࣜࢢࣜࢭࣜࢻࡢ㍺ ㏦ࡀ㜼ᐖࡉࢀࡿࡇ࡜࡟ࡼࡾ, ⫢⮚୰࡟ከᩘࡢ⬡⫫⁲ࡀほᐹࡉࢀࡿ[44, 45]ࠋAPAP ࡜ྠᵝ, ᅄሷ໬Ⅳ⣲ࡢ ⫢ẘᛶࡶᦤ㣵᮲௳ࡢᙳ㡪ࢆཷࡅࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠾ࡾ, 24 ᫬㛫⤯㣗ୗࡢࣛࢵࢺ࡛ࡣ⮬⏤ᦤ㣵ୗࡼࡾࡶ ᙉ࠸⫢㞀ᐖࡀほᐹࡉࢀ࡚࠸ࡿ[46]ࠋ ඛ⾜◊✲࡟ࡼࡾ, APAP ࡟ࡼࡿ⫢㞀ᐖࡣᢠ㓟໬⬟ࢆ᭷ࡍࡿ㣗ရ⏤᮶ࡢ≀㉁࡟ࡼࡾᢚไࡉࢀࡿࡇ࡜ࡀሗ࿌ ࡉࢀ࡚࠾ࡾ, ື≀ᐇ㦂࡟࡚, ࢺ࣐ࢺᢳฟ≀[48], ࢩࣙ࢘࢞⛉᳜≀㸦Zingiberaceae㸧ᢳฟ≀[49, 50], ࣈࢻ࢘ ✀Ꮚᢳฟ≀[51]ࡀ APAP ࡟ࡼࡿ⫢㞀ᐖࢆ㍍ῶࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋྠᵝ࡟, ࣅࢱ࣑ࣥ E [52]ࡸN -࢔ࢭࢳࣝࢩࢫࢸ࢖ࣥ[53]➼ࡢᢠ㓟໬≀㉁ࡢ௚, ࣜࣥࢦ࣏ࣜࣇ࢙ࣀ࣮ࣝ[54]ࡸS-࢔ࣜࣝࢩࢫࢸ࢖ࣥ[55]࡞࡝ ࡢ㣗ရ⏤᮶ࡢᢠ㓟໬ᡂศࡀᅄሷ໬Ⅳ⣲࡟ࡼࡿ⫢㞀ᐖ࡜⬡㉁㐣㓟໬ࢆᢚไࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋࡲ ࡓ, APAP ᢞ୚࡟ࡼࡿ⫢⣽⬊ቯṚ[33, 34]ࡸᅄሷ໬Ⅳ⣲ㄏⓎᛶࡢ⫢㞀ᐖ[56, 57]ࡣ, ⅖⑕཯ᛂࢆᢚไࡍࡿࡇ ࡜࡛㍍ῶࡉࢀࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗ࡚࠸ࡿࠋ࣏ࣉࣛࢱ࢖ࣉࡢࣉ࣏ࣟࣜࢫ࡛ࡣAPAP [14]ࡸᅄሷ໬Ⅳ⣲[15]

࡟ᑐࡍࡿಖㆤຠᯝࡢሗ࿌ࡀ࠶ࡿ୍᪉, EEBGP ࡟ࡘ࠸࡚ࡣ࡯࡜ࢇ࡝ሗ࿌ࡉࢀ࡚࠸࡞࠸ࠋEEBGP ࡣ, in vitro

࡟࠾࠸࡚ࣛࢪ࢝ࣝࢫ࢝࣋ࣥࢪ࣮ࣕ࡜ࡋ࡚άᛶ㓟⣲✀ࢆ┤᥋ᾘཤࡋ[11, 58, 59], ᢠ㓟໬㓝⣲ࢆㄏᑟࡍࡿࡇ ࡜ࡀ▱ࡽࢀ[11], ࣐࢘ࢫࡸࣛࢵࢺ࡟࠾࠸࡚ᢠ㓟໬㓝⣲άᛶࢆ㧗ࡵࡿࡇ࡜࡛ α-ࢼࣇࢳࣝ࢖ࢯࢳ࢜ࢩ࢔ࢿ࣮ ࢺ[19]ࡸỈᾐᣊ᮰ࢫࢺࣞࢫ[21]࡟ࡼࡿ⾑୰ࡢ⫢㞀ᐖ࣐࣮࣮࢝ࡢቑຍࢆపῶࡉࡏࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋ ࡲࡓ, EEBGP ࡣ⣽⬊ヨ㦂ࡸື≀ヨ㦂࡟࡚ᢠ⅖⑕స⏝ࢆ♧ࡍࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࡀ[60, 61], EEBGP ࡢ ᢠ⅖⑕స⏝࡜⫢㞀ᐖ࡜ࡢ㛵ಀ࡟ࡘ࠸࡚ࡣ᫂ࡽ࠿࡟࡞ࡗ࡚࠸࡞࠸ࠋ

ᮏ❶࡛ࡣ, ᐇ㦂 1 ࡟࠾࠸࡚, ప⏝㔞ࡢ APAP ᢞ୚࡟ࡼࡿ⫢യᐖࣔࢹࣝࢆ☜❧ࡍࡿࡓࡵ, APAP ࡸ NAPQI ࡢ௦ㅰ࡟㛵ࢃࡿUDP-glucuronosyltransferase 1A6 (Ugt1a6), sulfotransferase 1A1 (Sult1a1), Cyp2e1

15 ᦤ㣵᮲௳࡟ࡼࡿᙳ㡪ࢆ☜ㄆࡋࡓࠋ⥆࠸࡚, 800 mg/kg య㔜ࡢ APAP ࢆ⮬⏤ᦤ㣵ୗࡢᮅ᪉ࡲࡓࡣኤ᪉, ⤯㣗 ୗ, ࠶ࡿ࠸ࡣไ㝈⤥㣵㸦⮬⏤ᦤ㣵ୗࡢ 70%㸧᮲௳ୗࡢࣛࢵࢺ࡟ᢞ୚ࡋ, APAP ࡟ࡼࡿ⫢㞀ᐖࡢ᭷↓࠾ࡼࡧ ⛬ᗘ࡟ࡘ࠸᳨࡚ウࡋࡓࠋࡲࡓ, APAP ࡜ྠᵝ࡟ẘᛶⓎ⌧࡟ CYP2E1 ࡀ㛵ࢃࡿᅄሷ໬Ⅳ⣲࡟ࡘ࠸࡚, ⮬⏤ᦤ 㣵ୗ࠶ࡿ࠸ࡣ⤯㣗ୗࡢࣛࢵࢺ࡟ᢞ୚ࡋ, ⫢യᐖࡢ᭷↓࠾ࡼࡧ⛬ᗘ࡟ࡘ࠸᳨࡚ウࡋࡓࠋᐇ㦂 2 ࡛ࡣ, EEBGP ࡢ஦๓ᢞ୚ࡀ APAP ㄏⓎᛶ⫢㞀ᐖ࡟ᑐࡋ࡚ಖㆤⓗ࡟ാࡃ࠿ྰ࠿ࢆ᳨ウࡋ, ᢠ㓟໬㓝⣲࠾ࡼࡧ⅖⑕㛵㐃ᅉ Ꮚࡢ㑇ఏᏊⓎ⌧࡟ᑐࡍࡿᙳ㡪ࢆㄪ࡭ࡿࡇ࡜࡛, EEBGP ࡢᢠ㓟໬స⏝࠾ࡼࡧᢠ⅖⑕స⏝࡜⤌⧊യᐖ࡟ᑐࡍ ࡿಖㆤຠᯝ࡜ࡢ㛵ಀᛶ࡟ࡘ࠸᳨࡚ウࡋࡓࠋࡉࡽ࡟, ᐇ㦂 3 ࡛ EEBGP ࡢ஦๓ᢞ୚ࡀᅄሷ໬Ⅳ⣲ㄏⓎᛶ⫢ യᐖ࡟ᑐࡋ࡚ಖㆤⓗ࡟ാࡃ࠿ྰ࠿ࢆ᳨ウࡋࡓࠋ ࠙ᮦᩱ࡜᪉ἲࠚ ࣭⿕㦂≀㉁  ⿕㦂≀㉁࡟ࡣ, BGP ཎሢࡢ 95%࢚ࢱࣀ࣮ࣝᢳฟ≀㸦EEBGP; ࢔ࣆओ, ᒱ㜧, ᪥ᮏ㸧ࢆ⏝࠸ࡓࠋEEBGP ࢆࢹ࢟ࢫࢺࣜࣥ⢊ᮎ㸦ᯇ㇂໬Ꮫᕤᴗओ, රᗜ, ᪥ᮏ㸧࡜ 2:3 ࡢ๭ྜ࡛ΰྜࡋ࡚஝⇱ࡉࡏ, ୍⯡㣫⫱⏝ᅛᙧ 㣫ᩱ㸦CE-2; ᪥ᮏࢡࣞ࢔ओ, ᮾி, ᪥ᮏ㸧࡟ 0.3%㸦ᐇ㦂 2㸧࠶ࡿ࠸ࡣ 1.2%㸦ᐇ㦂 3㸧࡛ΰྜࡋࡓࠋ ࣭ື≀  ᐇ㦂࡟ࡣ, 5-6 㐌㱋ࡢ㞝ᛶ Wistar/ST ࣛࢵࢺ㸦᪥ᮏ SLCओ, 㟼ᒸ, ᪥ᮏ㸧ࢆ⏝࠸, ᐊ  23s1Υ, ‵ᗘ 55s10㸣, ᫂ᬯྛ 12 ᫬㛫㸦᫂ᮇ: 8:00㹼20:00㸧࡟ࢥࣥࢺ࣮ࣟࣝࡋࡓື≀ᐊ࡟࡚⣙ 1 㐌㛫㥆໬ࡉࡏࡓᚋ, ヨ 㦂࡟౑⏝ࡋࡓࠋື≀ヨ㦂ࡣ, ࢔ࣆओ㛗Ⰻᕝࣜࢧ࣮ࢳࢭࣥࢱ࣮ࡢື≀ᐇ㦂ጤဨ఍ࡢᢎㄆࡢࡶ࡜㸦ᐇ㦂 1: #42-002 ࠾ࡼࡧ#44-020, ᐇ㦂 2: #43-006, ᐇ㦂 3: #44-038㸧, ື≀೔⌮つᐃ࡟ᚑࡗ࡚ᐇ᪋ࡋࡓࠋ ᐇ ᐇ㦂1 ࣭⤥㣵᮲௳ ື≀ࢆ, ⮬⏤ᦤ㣵⩌, ๓᪥ 17:00 ࠿ࡽ 9:00 ࡲ࡛ࡢ 16 ᫬㛫⤯㣗⩌࡞ࡽࡧ࡟ไ㝈⤥㣵⩌࡟ศࡅ, CE-2 ࢆ୚ ࠼ࡓࠋไ㝈⤥㣵ࡣ, ඛ⾜ᩥ⊩ࢆཧ⪃࡟[62], ྠ⣔⤫, ྠ㐌㱋ࡢ㞝ᛶࣛࢵࢺ࡟࠾ࡅࡿ 1 ᪥࠶ࡓࡾࡢᖹᆒᦤ㣵 㔞ࡢ⣙70%࡟┦ᙜࡍࡿ 14 g ࢆẖ᪥ 17:00 ࡟⤥㣵ࡋ, 10 ᪥㛫⥅⥆ࡋࡓࠋไ㝈⤥㣵᮲௳ୗ࡛ࡣ,ẖ᪥ 9:00 ࡟ ୍⯡≧ែࢆほᐹࡋ, య㔜࠾ࡼࡧᦤ㣵㔞ࢆグ㘓ࡋࡓࠋ

16 ࣭㑇ఏᏊⓎ⌧㔞࠾ࡼࡧࢢࣝࢱࢳ࢜ࣥ㔞ࡢ ᐃ  ⮬⏤ᦤ㣵ୗ࠾ࡼࡧไ㝈⤥㣵᮲௳ୗ࡛ࡣ9:00 ࠾ࡼࡧ 16:00 ࡟, ⤯㣗ୗ࡛ࡣ 9:00 ࡟㸦ྛ n = 5㸧, ࢖ࢯࣇ ࣝࣛࣥ㸦࿴ග⣧⸆ᕤᴗओ, ኱㜰, ᪥ᮏ㸧྾ධ㯞㓉ୗ࡛⫢⮚ࢆ᦬ฟࡋ࡚㔜㔞ࢆ ᐃࡋࡓᚋ, ᾮయ❅⣲࡛෾⤖ ࡋ, ⸆≀௦ㅰ㓝⣲ࡢ㑇ఏᏊⓎ⌧㔞࠾ࡼࡧ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࢆ ᐃࡋࡓࠋ  ෾⤖ࡋࡓ⫢⮚⣙50 mg ࡟ ISOGEN®_{ϩ㸦ओࢽࢵ࣏ࣥࢪ࣮ࣥ, ᮾி, ᪥ᮏ㸧ࢆຍ࠼࡚ Polytron}® _PT1300D 㸦Kinematica AG, Lucerne, Switzerland㸧࡟࡚࣍ࣔࢪࢼ࢖ࢬ㸦12, 000 rpm㸧ࡋࡓࠋ⁛⳦ Milli-Q Ỉࢆຍ ࠼࡚, ᐊ ࡛ 15 ศ㛫㟼⨨ࡋࡓᚋ, 㐲ᚰศ㞳㸦12, 000™G, 4Υ, 15 ศ㸧ࡋ, ୖΎࢆᅇ཰ࡋࡓࠋᢳฟࡋࡓ Total RNA ࡣp-ࣈࣟࣔ࢔ࢽࢯ࣮ࣝ㸦࿴ග⣧⸆ᕤᴗओ㸧࠾ࡼࡧ 8 M ሷ໬ࣜࢳ࣒࢘⁐ᾮ㸦ࢼ࢝ࣛ࢖ࢸࢫࢡओ, ி㒔, ᪥ᮏ㸧࡟࡚⢭〇ࡋ, ᢳฟࡋࡓ Total RNA ࠿ࡽ SuperScript® _{III First-Strand Synthesis SuperMix for} qRT-PCR㸦Thermo Fisher Scientific, Waltham, MA, USA㸧ࢆ⏝࠸࡚ cDNA ࢆྜᡂࡋࡓࠋcDNA ࢆࢸࣥ ࣉ࣮ࣞࢺ࡜ࡋ, Applied Biosystems 7300 Real Time PCR system㸦Thermo Fisher Scientific㸧ࢆ⏝࠸࡚, ᐃ㔞PCR㸦95Υ, 15 ⛊Ѝ60Υ, 1 ศࢆ 40 ࢧ࢖ࢡࣝ㸧ࢆᐇ᪋ࡋࡓࠋReference gene ࡜ࡋ࡚ Glyceraldehyde- 3-phosphate dehydrogenase㸦Gapdh㸧㑇ఏᏊࢆ⏝࠸, ಶయࡈ࡜ࡢ㑇ఏᏊⓎ⌧㔞ࡣGapdh㑇ఏᏊࡢ┦ᑐ 㔞࡜ࡋ࡚⟬ฟࡋࡓࠋ ᐃࡍࡿ⸆≀௦ㅰ㓝⣲㑇ఏᏊࡣ, ᩥ⊩᝟ሗࢆᇶ࡟[24-26, 63, 64], APAP ௦ㅰ࡟㛵୚ࡍ ࡿUgt1a6, Sult1a1, Cyp2e1࠾ࡼࡧGstm1࡜ࡋࡓࠋ౑⏝ࡋࡓࣉࣛ࢖࣐࣮ࢆ⾲2-1 ࡟♧ࡋࡓࠋ

 ෾⤖ࡋࡓ⫢⮚⣙100 mg ࡟ 5% (w/v) ࢫࣝ࣍ࢧࣜࢳࣝ㓟⁐ᾮ㸦ࢼ࢝ࣛ࢖ࢸࢫࢡओ㸧ࢆຍ࠼࡚࣍ࣔࢪࢼ࢖ ࢬ㸦12, 000 rpm㸧ࡋࡓࠋ㐲ᚰศ㞳㸦8, 000™G, 4Υ, 10 ศ㸧ᚋࡢୖΎࢆ⏝࠸, GSSG/GSH Quantification Kit㸦ओྠோ໬Ꮫ◊✲ᡤ, ⇃ᮏ, ᪥ᮏ㸧࡟࡚⥲ࢢࣝࢱࢳ࢜ࣥ㔞ࢆ ᐃࡋࡓࠋಶయࡈ࡜ࡢࢢࣝࢱࢳ࢜ࣥ㔞ࡣ ⫢⮚‵㔜㔞࡛᥮⟬ࡋ, ⫢⮚࠶ࡓࡾࡢࢢࣝࢱࢳ࢜ࣥ㔞࡜ࡋ࡚⟬ฟࡋࡓࠋ ࣭APAP ㄏⓎᛶ⫢㞀ᐖࡢホ౯  APAP ࡢᢞ୚㔞ࡣ, ࣄࢺ࡟࠾࠸࡚⫢㞀ᐖࢆ♧ࡍ 8 g ࢆࣛࢵࢺࡢయ⾲㠃✚࡛᥮⟬ࡋࡓ 800 mg/kg య㔜࡜ࡋ ࡓࠋAPAP㸦ࢼ࢝ࣛ࢖ࢸࢫࢡओ㸧ࢆ 0.5% (w/v)࣓ࢳࣝࢭ࣮ࣝࣟࢫ㸦ಙ㉺໬Ꮫᕤᴗओ, ᮾி, ᪥ᮏ㸧⁐ᾮ࡟ ᠱ⃮ࡋ, ⮬⏤ᦤ㣵ୗ࡛ࡣ 9:00㸦n = 3㸧࠾ࡼࡧ 16:00㸦n = 5㸧, ⤯㣗ୗ࡛ࡣ 9:00㸦n = 5㸧, ไ㝈⤥㣵᮲௳ ୗ࡛ࡣ16:00㸦n = 5㸧࡟, ࡑࢀࡒࢀ༢ᅇᙉไ⤒ཱྀᢞ୚ࡋࡓࠋ࡞࠾, ⮬⏤ᦤ㣵ୗ 16:00 ᢞ୚࡛ࡣ, APAP ᢞ ୚࠿ࡽ๗᳨ࡲ࡛⤯㣗ࡋ, ⮬⏤ᦤ㣵ୗ 9:00 ᢞ୚࠾ࡼࡧ⤯㣗ୗ 9:00 ᢞ୚࡛ࡣ 10:00 ࠿ࡽ 16:00 ࡲ࡛⮬⏤ᦤ㣵

17 ࡜ࡋ, 16:00 ࠿ࡽ๗᳨ࡲ࡛ࡣ⤯㣗ࡋࡓࠋไ㝈⤥㣵᮲௳ୗ 16:00 ᢞ୚࡛ࡣ 17:00 ࡟⣙ 14 g ⤥㣵ࡋࡓࠋ ࠸ࡎࢀࡢື≀ࡶAPAP ᢞ୚࠿ࡽ⣙ 24 ᫬㛫ᚋ࡟࢖ࢯࣇࣝࣛࣥ྾ධ㯞㓉ୗ࡛⭡኱㟼⬦ࡼࡾ᥇⾑ࡋࡓᚋ, ᨺ ⾑⮴Ṛࡋࡓࠋ⫢⮚ࢆ᦬ฟࡋࡓᚋ, 10%୰ᛶ⦆⾪࣐࣍ࣝࣜࣥ⁐ᾮ㸦࣐࢖ࣝࢻ࣒࣍ࣝ®_{10N; ࿴ග⣧⸆ᕤᴗओ㸧} ࡟࡚ᅛᐃࡋ, ᐃἲ࡟ᚑࡗ࡚࣐࣊ࢺ࢟ࢩࣜࣥ-࢚࢜ࢪࣥ㸦H.E.㸧ᰁⰍᶆᮏࢆస〇ࡋ, ගᏛ㢧ᚤ㙾࡟࡚ほᐹࡋ ࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣ, ⾑Ύࢆ㐲ᚰศ㞳㸦3, 000 rpm, 4Υ, 15 ศ㸧ࡋࡓᚋ, ⏕໬Ꮫ⮬ືศᯒ⿦⨨㸦ᐩኈࢻࣛ ࢖ࢣ࣒ 7000i; ᐩኈࣇ࢕࣒࣓ࣝࢹ࢕࢝ࣝओ, ᮾி, ᪥ᮏ㸧ࢆ⏝࠸࡚, ࢔ࣛࢽࣥ࢔࣑ࣀࢺࣛࣥࢫࣇ࢙࣮ࣛࢮ 㸦ALT㸧್࠾ࡼࡧ࢔ࢫࣃࣛࢠࣥ㓟࢔࣑ࣀࢺࣛࣥࢫࣇ࢙࣮ࣛࢮ㸦AST㸧್ࢆ ᐃࡋࡓࠋ ࣭ᅄሷ໬Ⅳ⣲ㄏⓎᛶ⫢㞀ᐖࡢホ౯  ᅄሷ໬Ⅳ⣲ࡢᢞ୚㔞ࡣ800 mg/kg య㔜࡜ࡋࡓࠋᅄሷ໬Ⅳ⣲㸦࿴ග⣧⸆ᕤᴗओ㸧ࢆ 0.1% (v/v) Tween 80 㸦ᮾி໬ᡂᕤᴗओ, ᮾி, ᪥ᮏ㸧ࢆྵࡴ 0.5% (w/v)࣓ࢳࣝࢭ࣮ࣝࣟࢫ⁐ᾮ࡟࣍ࣔࢪࢼ࣮ࢨ࣮㸦TAITEC CULTRAS HOMOGENIZER VP-60S; ࢱ࢖ࢸࢵࢡओ, ᇸ⋢, ᪥ᮏ㸧ࢆ⏝࠸࡚ศᩓࡉࡏ, ⮬⏤ᦤ㣵ୗ㸦n = 5㸧࠶ࡿ࠸ࡣ 16 ᫬㛫⤯㣗ୗ㸦n = 4㸧ࡢ 10:00 ࡟, ࡑࢀࡒࢀ༢ᅇᙉไ⤒ཱྀᢞ୚ࡋࡓࠋ࡞࠾, ⮬⏤ᦤ㣗ୗᢞ ୚࡛ࡣ, ᢞ୚᪥ࡢ 8:00 ࡲ࡛ࡣ⮬⏤࡟ᦤ㣗ࡉࡏ, ᢞ୚ᚋࡣ 2 ᫬㛫⤯㣗ࡉࡏࡓࠋ⤯㣗ୗᢞ୚࡛ࡣ, ᢞ୚๓᪥ࡢ 18:00 ࠿ࡽ⤯㣗ࡉࡏ, ᅄሷ໬Ⅳ⣲ᢞ୚ᚋ 2 ᫬㛫ࡲ࡛⤯㣗ࢆ⥅⥆ࡋࡓࠋࡑࡢᚋ, ࠸ࡎࢀࡢ᮲௳࡛ࡶ⮬⏤࡟ᦤ 㣗ࡉࡏ, ᢞ୚᪥ࡢ 17:00 ࠿ࡽ๗᳨ࡲ࡛ࡣ⤯㣗ࡉࡏࡓࠋ ࠸ࡎࢀࡢື≀ࡶᅄሷ໬Ⅳ⣲ᢞ୚࠿ࡽ⣙24 ᫬㛫ᚋ࡟࢖ࢯࣇࣝࣛࣥ྾ධ㯞㓉ୗ࡛⭡኱㟼⬦ࡼࡾ᥇⾑ࡋࡓᚋ, ᨺ⾑⮴Ṛࡋࡓࠋ⫢⮚ࢆ᦬ฟࡋࡓᚋ, 10%୰ᛶ⦆⾪࣐࣍ࣝࣜࣥ⁐ᾮ࡟࡚ᅛᐃࡋ, ᐃἲ࡟ᚑࡗ࡚ H.E.ᰁⰍᶆᮏ ࢆస〇ࡋ, ගᏛ㢧ᚤ㙾࡟࡚ほᐹࡋࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣ, ⾑Ύࢆ㐲ᚰศ㞳㸦3, 000 rpm, 4Υ, 15 ศ㸧ࡋࡓᚋ, ⏕໬Ꮫ⮬ືศᯒ⿦⨨㸦ᐩኈࢻࣛ࢖ࢣ࣒7000i㸧ࢆ⏝࠸࡚, ALT ್࠾ࡼࡧ AST ್ࢆ ᐃࡋࡓࠋ

ᐇ ᐇ㦂2

࣭ᐇ㦂ࢹࢨ࢖ࣥ

Control ⩌࡟ࡣ CE-2 ࢆ, EEBGP ⩌ࡣ 0.3% EEBGP 㣗ࢆ 7 ᪥㛫㐃⥆࡛୚࠼ࡓࠋAPAP ࡢᢞ୚㔞࠾ࡼࡧ ᢞ୚᫬㛫ࡣᐇ㦂1 ࡢ⤖ᯝ࠿ࡽタᐃࡋ, ᢞ୚㛤ጞ࠿ࡽ 8 ᪥┠ࡢ 16:00 ࡟, 0.5% (w/v)࣓ࢳࣝࢭ࣮ࣝࣟࢫ⁐ᾮ ࡟ᠱ⃮ࡋࡓAPAP ࢆ 800 mg/kg య㔜࡛༢ᅇᙉไ⤒ཱྀᢞ୚ࡋ, ๗᳨ࡲ࡛⤯㣗ࡉࡏࡓࠋᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ ቯṚࡀࡳࡽࢀࡓControl ⩌ 4 ౛࠾ࡼࡧ EEBGP ⩌ 5 ౛ࢆホ౯࡟⏝࠸ࡓࠋ

18

APAP ᢞ୚࠿ࡽ⣙ 24 ᫬㛫ᚋ࡟࢖ࢯࣇࣝࣛࣥ྾ධ㯞㓉ୗ࡛⭡኱㟼⬦⾑ࢆᚓࡓᚋ, ᒕẅࡋ, ⫢⮚ࢆ᦬ฟࡋ ࡓࠋ⫢⮚ࡢ୍㒊ࡣᾮయ❅⣲࡛෾⤖ࡋ, ᢠ㓟໬㓝⣲࠾ࡼࡧ⅖⑕㛵㐃㑇ఏᏊࡢⓎ⌧㔞ࢆ ᐃࡋࡓࠋ⫢⮚ࡢṧࡾ

ࡢ㒊ศࡣ 10%୰ᛶ⦆⾪࣐࣍ࣝࣜࣥ⁐ᾮ࡛ᅛᐃࡋࡓᚋ, ᐃἲ࡟ᚑࡗ࡚ H.E.ᰁⰍᶆᮏࢆస〇ࡋ, ගᏛ㢧ᚤ㙾

࡟࡚ほᐹࡋࡓࠋ⫢⮚ࡢṇᖖ㡿ᇦ࡟༨ࡵࡿቯṚ㡿ᇦࡢ๭ྜࡣ, 㢧ᚤ㙾㸦™50 ಸ㸧ୗ࡛ࣛࣥࢲ࣒࡟ 8 ᯛྲྀᚓ ࡋࡓ⏬ീࢆ⏝࠸࡚㠃✚್ࢆ⟬ฟࡋࡓࠋ⏬ീゎᯒ࡟ࡣImage J㸦National Institutes of Health, USA㸧౑ ⏝ࡋࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣ, ⾑Ύࢆ㐲ᚰศ㞳㸦3, 000 rpm, 4Υ, 15 ศ㸧ࡋࡓᚋ, ⏕໬Ꮫ⮬ືศᯒ⿦⨨ࢆ⏝࠸ ࡚, ALT ್࠾ࡼࡧ AST ್ࢆ ᐃࡋࡓࠋ

࣭ᐃ㔞PCR

 ᢠ㓟໬㓝⣲࡛࠶ࡿSod1, Sod2, Catalase㸦Cat㸧, Gpx1, Gpx4, Peroxiredoxin-1㸦Prdx1㸧, Glutathione reductase㸦Gsr㸧, ࢧ࢖ࢺ࢝࢖࡛ࣥ࠶ࡿIl1b, Il6, Il10, Il23a, Tnf, ࢣࣔ࢝࢖࡛ࣥ࠶ࡿCcl2, Ccl3, CXC motif chemokine ligand-2㸦Cxcl2㸧࠾ࡼࡧNitric oxide synthase-2㸦Nos2㸧㑇ఏᏊࡢⓎ⌧㔞ࢆᐃ㔞PCR ࡟࡚ ᐃࡋࡓࠋControl ⩌࠾ࡼࡧ EEBGP ⩌ࡼࡾᚓࡽࢀࡓ⫢⮚⣙ 50 mg ࡟ ISOGEN®_{ϩࢆຍ࠼࡚࡟࡚࣍} ࣔࢪࢼ࢖ࢬ㸦12, 000 rpm㸧ࡋ, ᐇ㦂 1 ࡢ᪉ἲ࡟ᚑࡗ࡚ cDNA ࢆྜᡂࡋࡓᚋ, ᐃ㔞 PCR ࡟࡚ᢠ㓟໬㓝⣲ ࠾ࡼࡧ⅖⑕㛵㐃㑇ఏᏊࡢⓎ⌧㔞ࢆ ᐃࡋࡓࠋ౑⏝ࡋࡓࣉࣛ࢖࣐࣮ࢆ⾲2-2 ࡟♧ࡋࡓࠋReference gene ࡟ ࡣGapdh㑇ఏᏊ㸦⾲2-1㸧ࢆ⏝࠸࡚, ಶయࡈ࡜ࡢ㑇ఏᏊⓎ⌧㔞ࡣGapdh㑇ఏᏊࡢ┦ᑐ㔞࡜ࡋ࡚⟬ฟࡋࡓࠋ ᐇ ᐇ㦂3 ࣭ᐇ㦂ࢹࢨ࢖ࣥ

Control ⩌࡟ࡣ CE-2 ࢆ, EEBGP ⩌ࡣ 1.2% EEBGP 㣗ࢆ 7 ᪥㛫㐃⥆࡛୚࠼ࡓࠋᢞ୚㛤ጞ࠿ࡽ 8 ᪥┠ ࡢ 9:00 ࡟, 0.1% (w/v) Tween 80 ྵ᭷ 0.5% (w/v)࣓ࢳࣝࢭ࣮ࣝࣟࢫ⁐ᾮ࡟ᠱ⃮ࡋࡓᅄሷ໬Ⅳ⣲ࢆ 800 mg/kg య㔜࡛༢ᅇᙉไ⤒ཱྀᢞ୚ࡋࡓࠋࡑࡢᚋ, ࠸ࡎࢀࡢ᮲௳࡛ࡶ⮬⏤࡟ᦤ㣗ࡉࡏ, ᢞ୚᪥ࡢ 17:00 ࠿ࡽ๗ ᳨ࡲ࡛ࡣ⤯㣗ࡉࡏࡓࠋᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚࡀࡳࡽࢀࡓྛ⩌6 ౛ࢆホ౯࡟⏝࠸ࡓࠋ ᅄሷ໬Ⅳ⣲ᢞ୚࠿ࡽ⣙24 ᫬㛫ᚋ࡟࢖ࢯࣇࣝࣛࣥ྾ධ㯞㓉ୗ࡛⭡኱㟼⬦⾑ࢆᚓࡓᚋ, ᒕẅࡋ, ⫢⮚ࢆ᦬ ฟࡋ࡚ 10%୰ᛶ⦆⾪࣐࣍ࣝࣜࣥ⁐ᾮ࡛ᅛᐃࡋࡓᚋ, ᐃἲ࡟ᚑࡗ࡚ H.E.ᰁⰍᶆᮏࢆస〇ࡋ, ගᏛ㢧ᚤ㙾࡟ ࡚ほᐹࡋࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣ, ⾑Ύࢆ㐲ᚰศ㞳㸦3, 000 rpm, 4Υ, 15 ศ㸧ࡋࡓᚋ, ⏕໬Ꮫ⮬ືศᯒ⿦⨨ࢆ ⏝࠸࡚, ALT ࠾ࡼࡧ AST ࢆ ᐃࡋࡓࠋ

19 ࣭⤫ィฎ⌮

 ᐇ㦂1 ࡛ࡣ, ⸆≀௦ㅰ㓝⣲ࡢ㑇ఏᏊⓎ⌧㔞, ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞, APAP ᢞ୚ 24 ᫬㛫ᚋࡢ⾑୰ ALT ್ ࠾ࡼࡧAST ್ࡣ, Bartlett ࡢ᳨ᐃ࡟࡚ྛ⩌ࡢศᩓࢆẚ㍑ࡋ, ➼ศᩓࡢሙྜࡣ Dunnett ࡢከ㔜ẚ㍑᳨ᐃ, ୙ ➼ศᩓࡢሙྜࡣSteel ࡢከ㔜ẚ㍑᳨ᐃࢆ⏝࠸࡚⮬⏤ᦤ㣵ୗ 9:00 ࡜ࡑࡢ௚ࡢ᮲௳ୗ࡛ẚ㍑ࡋࡓࠋຍ࠼࡚, ไ 㝈⤥㣵᮲௳ୗࡢ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࡣ, F ᳨ᐃ࡟࡚ศᩓࢆẚ㍑ࡋࡓᚋ, Student ࡢ t ᳨ᐃࢆ⏝࠸࡚, 2 ࡘࡢ ࢱ࢖࣒࣏࢖ࣥࢺ㸦9:00 ࠾ࡼࡧ 16:00㸧㛫࡛ẚ㍑ࡋࡓࠋᅄሷ໬Ⅳ⣲ᢞ୚ 24 ᫬㛫ᚋࡢ⾑୰ ALT ್࠾ࡼࡧ AST ್ࡣ, F ᳨ᐃ࡟࡚ศᩓࢆẚ㍑ࡋࡓᚋ, Welch ࡢ t ᳨ᐃࢆ⏝࠸࡚⮬⏤ᦤ㣵ୗ࡜⤯㣗ୗ࡛ẚ㍑ࡋࡓࠋᐇ㦂 2 ࡛ ࡣ, ⾑୰ ALT ್࠾ࡼࡧ AST ್, ⫢⣽⬊ቯṚ㡿ᇦࡢ๭ྜ, ᢠ㓟໬㓝⣲࠾ࡼࡧ⅖⑕㛵㐃㑇ఏᏊࡢⓎ⌧㔞ࡣ, F ᳨ᐃ࡟࡚ศᩓࢆẚ㍑ࡋࡓᚋ, ➼ศᩓࡢሙྜࡣ Student ࡢ t ᳨ᐃ, ୙➼ศᩓࡢሙྜࡣ Welch ࡢ t ᳨ᐃࢆ⏝࠸ ࡚Control ⩌࡜ EEBGP ⩌࡛ẚ㍑ࡋࡓࠋ⅖⑕㛵㐃㑇ఏᏊࡢⓎ⌧㔞࡜⫢⣽⬊ቯṚ㡿ᇦࡢ๭ྜࡣ, Spearman ࡢ㡰఩┦㛵ಀᩘࢆ⏝࠸࡚┦㛵ࡢ᭷↓ࢆ☜ㄆࡋࡓࠋᐇ㦂3 ࡛ࡣ, ⾑୰ ALT ್࠾ࡼࡧ AST ್ࡣ, F ᳨ᐃ࡟࡚ ศᩓࢆẚ㍑ࡋࡓᚋ, Student ࡢ t ᳨ᐃࢆ⏝࠸࡚ Control ⩌࡜ EEBGP ⩌࡛ẚ㍑ࡋࡓࠋ⤫ィฎ⌮࡟ࡣ࢚ࢡࢭ ࣝ⤫ィ2012㸦Social Survey Reserch Information Co., Ltd, ᮾி, ᪥ᮏ㸧ࢆ⏝࠸, ᭷ពỈ‽ࡣp < 0.05 ࡜ࡋࡓࠋ ࠙⤖ᯝࠚ ᐇ ᐇ㦂1 ࣭ไ㝈⤥㣵᮲௳ୗ࡟࠾ࡅࡿ୍⯡≧ែ, య㔜, ᦤ㣵㔞  ไ㝈⤥㣵⩌ࡢ࠸ࡎࢀࡢື≀࡟࠾࠸࡚ࡶ୍⯡≧ែ࡟␗ᖖࡣࡳࡽࢀ࡞࠿ࡗࡓࠋไ㝈⤥㣵᮲௳ୗ࡛㣫⫱ࡋࡓ ື≀࡟య㔜పୗࡣࡳࡽࢀࡎ㸦ᅗ2-3㸧, ⮬⏤ᦤ㣵⩌ࡢ 83%࡛࠶ࡗࡓࠋไ㝈⤥㣵⩌࡛ࡣ, ඲࡚ࡢಶయࡀヨ㦂 ᮇ㛫ࢆ㏻ࡋ࡚୚࠼ࡓ඲㔞㸦14 g㸧ࢆᦤ㣗ࡋࡓࠋ ࣭␗࡞ࡿᦤ㣵᮲௳ୗ࡟࠾ࡅࡿ⸆≀௦ㅰ㓝⣲㑇ఏᏊࡢⓎ⌧㔞࠾ࡼࡧ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࡢኚ໬ Cyp2e1 㑇ఏᏊࡢⓎ⌧㔞ࡣ, ⮬⏤ᦤ㣵ୗ 9:00 ࡜ẚ㍑ࡋ࡚⮬⏤ᦤ㣵ୗ 16:00 ࡛ࡣ᭷ព࡞ኚ໬ࡀࡳࡽࢀ࡞ ࠿ࡗࡓࡶࡢࡢ, ⤯㣗ୗ࠾ࡼࡧไ㝈⤥㣵᮲௳ୗ࡛ࡣ᭷ព࡟ቑຍࡋࡓ㸦p < 0.01, ᅗ 2-4㸧ࠋ⮬⏤ᦤ㣵ୗ 16:00, ⤯㣗ୗ࠾ࡼࡧไ㝈⤥㣵᮲௳ୗࡢUgt1a6, Sult1a1࠾ࡼࡧGstm1㑇ఏᏊࡢⓎ⌧㔞ࡣ, ⮬⏤ᦤ㣵ୗ 9:00 ࡜ẚ ㍑ࡋ࡚᭷ព࡞ኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦ᅗ2-4㸧ࠋ

20 ⮬⏤ᦤ㣵ୗ16:00 ࡢ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࡣ, ⮬⏤ᦤ㣵ୗ 9:00 ࡜ẚ㍑ࡋ࡚᭷ព࡞ኚ໬ࡀࡳࡽࢀ࡞࠿ࡗࡓࠋ ไ㝈⤥㣵᮲௳ୗ9:00 ࡛ࡣ⮬⏤ᦤ㣵ୗ 9:00 ࡢ 86%࡜ࢃࡎ࠿࡞ῶᑡࡀࡳࡽࢀࡓࡀ, ᭷ព࡞ኚ໬࡛ࡣ࡞࠿ࡗࡓࠋ ୍᪉, ⤯㣗ୗ࠾ࡼࡧไ㝈⤥㣵ୗ 16:00 ࡛ࡣ⮬⏤ᦤ㣵ୗ 9:00 ࡜ẚ㍑ࡋ࡚᭷ព࡟పୗࡋ㸦p < 0.01㸧, ຍ࠼࡚ ไ㝈⤥㣵ୗ16:00 ࡛ࡣไ㝈⤥㣵ୗ 9:00 ࡜ẚ㍑ࡋ࡚᭷ព࡟పୗࡋࡓ㸦p < 0.01, ᅗ 2-5㸧ࠋ ࣭APAP ㄏⓎᛶ⫢㞀ᐖ࡟ᑐࡍࡿᦤ㣵᮲௳ࡢᙳ㡪  ྛ᮲௳ୗ࡟࠾࠸࡚, 800 mg/kg య㔜ࡢ APAP ࢆ⤒ཱྀᢞ୚ࡋࡓ࡜ࡇࢁ, ⮬⏤ᦤ㣵ୗ 9:00 ࡢᢞ୚࡛ࡣ, ࠸ࡎ ࢀࡢື≀ࡢ⫢⮚࡟ࡶ␗ᖖࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦ᅗ2-6A㸧ࠋ୍᪉, ⮬⏤ᦤ㣵ୗ 16:00 ᢞ୚࡛ࡣ, 4 ౛࡛⅖⑕⣽ ⬊ᾐ₶ࢆకࡗࡓᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀ, ูࡢ 1 ౛࡛୰ᚰ㟼⬦࿘ᅖ࡟⅖⑕⣽⬊ᾐ₶ࢆకࡗࡓ ༢⣽⬊ᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀࡓ㸦ᅗ2-6B㸧ࠋᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚࡀࡳࡽࢀࡓಶయ࡛ࡣ, ALT ್ࡀ

⮬⏤ᦤ㣵ୗࡢᖹᆒ್㸦51.0s9.2 U/L㸧ࡢ⣙ 160%௨ୖ࡟, AST ್ࡀ⮬⏤ᦤ㣵ୗࡢᖹᆒ್㸦84.7s8.0 U/L㸧

࡟ᑐࡋ࡚300%௨ୖ࡟ቑຍࡋ, ༢⣽⬊ᛶࡢ⫢⣽⬊ቯṚࡀࡳࡽࢀࡓಶయ࡛ࡣ, AST ್ࡀ⮬⏤ᦤ㣵ୗࡢᖹᆒ್ ࡢ⣙150%࡟ቑຍࡋࡓࡀ, ALT ್ࡣ⮬⏤ᦤ㣵ୗ࡜ኚࢃࡽ࡞࠿ࡗࡓ㸦⾲ 2-3㸧ࠋ ⤯㣗ୗࡢᢞ୚࡛ࡣ, 2 ౛࡛⅖⑕⣽⬊ᾐ₶ࢆకࡗࡓᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀ㸦ᅗ 2-6C㸧, ูࡢ 2 ౛࡛୰ᚰ㟼⬦࿘ᅖ࡟⅖⑕⣽⬊ᾐ₶ࢆకࡗࡓ༢⣽⬊ᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀࡓࠋᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯ Ṛࡀࡳࡽࢀࡓಶయ࡛ࡣ, ALT ್ࡀ⮬⏤ᦤ㣵ୗࡢᖹᆒ್ࡢ⣙ 150%࡟, AST ್ࡀ⮬⏤ᦤ㣵ୗࡢᖹᆒ್࡟ᑐࡋ ࡚ 300%௨ୖ࡟ቑຍࡋ, ༢⣽⬊ᛶࡢ⫢⣽⬊ቯṚࡀࡳࡽࢀࡓಶయ࡛ࡣ, AST ್ࡀ⮬⏤ᦤ㣵ୗࡢᖹᆒ್ࡢ⣙ 140%࡟ቑຍࡋࡓࡀ, ALT ್ࡣ⮬⏤ᦤ㣵ୗ࡜ኚࢃࡽ࡞࠿ࡗࡓ㸦⾲ 2-3㸧ࠋ  ไ㝈⤥㣵᮲௳ୗ࡛ࡣ, 3 ౛࡛୰ᚰ㟼⬦࿘ᅖ࡟⅖⑕⣽⬊ᾐ₶ࢆకࡗࡓ༢⣽⬊ᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀ 㸦ᅗ2-6D㸧, ࡇࢀࡽࡢື≀࡛ࡣ AST ್ࡀ⮬⏤ᦤ㣵ୗࡢᖹᆒ್ࡢ⣙ 120%࡟ቑຍࡋࡓࡀ, ALT ್ࡣ඲౛࡛ ኚࢃࡽ࡞࠿ࡗࡓ㸦⾲2-3㸧ࠋ  ࠸ࡎࢀࡢ᮲௳࡛ࡶ, ALT ್࠾ࡼࡧ AST ್ࡣ⮬⏤ᦤ㣵ୗ 9:00 ࡜ẚ㍑ࡋ࡚᭷ព࡞ኚ໬࡛ࡣ࡞࠿ࡗࡓࠋ ࣭ᅄሷ໬Ⅳ⣲ㄏⓎᛶ⫢㞀ᐖ࡟ᑐࡍࡿᦤ㣵᮲௳ࡢᙳ㡪  ⮬⏤ᦤ㣵ୗ࡛800 mg/kg య㔜ࡢᅄሷ໬Ⅳ⣲ࢆ⤒ཱྀᢞ୚ࡋࡓ࡜ࡇࢁ, ඲౛࡛⅖⑕ᛶ⣽⬊ᾐ₶ࢆకࡗࡓᑠ ⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚ࡜, ୰㛫ᇦ࡛⫢⣽⬊ࡢ⬡⫫ኚᛶࡀほᐹࡉࢀࡓ㸦ᅗ 2-7A-a㸧ࠋ⤯㣗ୗࡢᢞ୚࡛ࡣ, ඲

21

౛࡛ᑠⴥ୰ᚰᇦ࠿ࡽ୰㛫ᇦ࡟࠿ࡅ࡚⅖⑕ᛶ⣽⬊ᾐ₶ࢆకࡗࡓ⫢⣽⬊ቯṚࡀࡳࡽࢀ, ⮬⏤ᦤ㣵ୗ࡟ẚ࡭࡚ ᑡᩘ࡛ࡣ࠶ࡿࡀ, ୰㛫ᇦ࡟⬡⫫ኚᛶࢆ♧ࡍ⫢⣽⬊ࡀほᐹࡉࢀࡓ㸦ᅗ 2-7A-b㸧ࠋ⫢⣽⬊ቯṚࡢ⠊ᅖ࡜┦㛵ࡍ ࡿࡼ࠺࡟, ALT ್࡛ࡣ᭷ព࡞ᕪ࡛ࡣ࡞࠿ࡗࡓࡶࡢࡢ, ⮬⏤ᦤ㣵ୗ㸦130.6s104.1 U/L㸧࡟ᑐࡋ, ⤯㣗ୗ㸦1, 918.5s598.7 U/L㸧࡛㧗ࡃ㸦p = 0.05㸧, AST ್ࡣ⮬⏤ᦤ㣵ୗࡀ 310.0s252.1 U/L ࡛࠶ࡗࡓࡢ࡟ᑐࡋ, ⤯ 㣗ୗ࡛ࡣ3, 899.3s1150.3 U/L ࡜᭷ព࡟㧗࠿ࡗࡓ㸦p < 0.05, ᅗ 2-7B㸧ࠋ ᐇ ᐇ㦂2 ୍࣭⯡≧ែ, య㔜, ᦤ㣵㔞  EEBGP ࡢ 7 ᪥㛫ΰ㣵ᢞ୚࡟ࡼࡿ୍⯡≧ែࡢ␗ᖖࡣࡳࡽࢀࡎࠊయ㔜᥎⛣ࠊయ㔜ቑຍ㔞࠾ࡼࡧᦤ㣵㔞ࡣ ᑐ↷⩌࡜ኚࢃࡽ࡞࠿ࡗࡓ㸦ࢹ࣮ࢱ♧ࡉࡎ㸧ࠋ ࣭⑓⌮⤌⧊ホ౯࠾ࡼࡧ⏕໬Ꮫ᳨ᰝ  APAP ࢆᢞ୚ࡋࡓࣛࢵࢺࡢ⫢⮚ࢆ⑓⌮⤌⧊Ꮫⓗ࡟ほᐹࡋࡓ࡜ࡇࢁ, ⅖⑕ᛶ⣽⬊ࡢᾐ₶ࢆకࡗࡓᑠⴥ୰ ᚰᛶࡢ⫢⣽⬊ቯṚࡀࡳࡽࢀࡓ㸦ᅗ2-8A-a㸧ࠋEEBGP ࢆᢞ୚ࡋࡓື≀࡛ࡣ, Control ⩌ࡢື≀࡜ྠᵝ࡟⅖ ⑕ᛶ⣽⬊ࡢᾐ₶ࢆకࡗࡓᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚࡀࡳࡽࢀࡓࡀ㸦ᅗ2-8A-b㸧, EEBGP ⩌ࡢ⫢⣽⬊ቯṚࡢ ⠊ᅖ㸦28.2s5.0%㸧ࡣ Control ⩌㸦45.9s5.3%㸧࡜ẚ㍑ࡋ࡚᭷ព࡟పୗࡋࡓ㸦p < 0.05, ᅗ 2-8B㸧ࠋࡲࡓ, ⫢⣽⬊ቯṚࡢኚ໬࡜ྠᵝ࡟, Control ⩌࡜ EEBGP ⩌ࡢ㛫࡟᭷ពᕪࡣࡳࡽࢀ࡞࠿ࡗࡓࡶࡢࡢ, EEBGP ᢞ୚ ࡟ࡼࡾALT ್㸦256.4s69.2 U/L㸧ࡣ Control ⩌㸦803.8s341.1 U/L㸧ࡢ⣙ 1/3 ࡟, AST ್㸦1, 807.6s 738.3 U/L㸧ࡣ Control ⩌㸦3, 447.3s998.8 U/L㸧ࡢ⣙༙ศ࡟పୗࡋࡓ㸦ᅗ 2-8C㸧ࠋ

࣭㑇ఏᏊⓎ⌧㔞ࡢኚ໬

Sod1, Sod2, Cat, Gpx1, Gpx4, Prdx1, Gsrࡢ࠸ࡎࢀࡢᢠ㓟໬㓝⣲㑇ఏᏊࡢⓎ⌧㔞࡟࠾࠸࡚ࡶ, EEBGP ⩌࡜Control ⩌ࡢ㛫࡛⤫ィⓗ࡟᭷ព࡞ኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦ᅗ 2-9㸧ࠋ

୍᪉, ⅖⑕㛵㐃㑇ఏᏊ࡛ࡣ, Control ⩌࡜ẚ㍑ࡋ࡚, EEBGP ⩌࡛Il10㑇ఏᏊࡢⓎ⌧㔞ࡀ᭷ព࡟పୗࡋ㸦p < 0.05㸧, ᭷ព࡞ᕪࡣࡳࡽࢀ࡞࠿ࡗࡓࡶࡢࡢ, Il1b㑇ఏᏊࡢⓎ⌧㔞ࡢపୗഴྥࡀࡳࡽࢀࡓ㸦p = 0.06㸧㸦ᅗ 2-10㸧ࠋࡑࡢ௚ࡢ࠸ࡎࢀࡢ㑇ఏᏊⓎ⌧㔞࡟࠾࠸࡚ࡶ, EEBGP ⩌࡜ Control ⩌ࡢ㛫࡛⤫ィⓗ࡟᭷ព࡞ኚ໬ࡣ ࡳࡽࢀ࡞࠿ࡗࡓࡀ, EEBGP ⩌࡛ࡣIl6࠾ࡼࡧCxcl2㑇ఏᏊࡢⓎ⌧㔞ࡀࡑࢀࡒࢀControl ⩌ࡢ 57.2%࠾ࡼ

22 ࡧ46.0%࡟పୗࡋࡓ㸦ᅗ 2-10㸧ࠋ ⫢⣽⬊ቯṚࡢ⠊ᅖ࡜㑇ఏᏊⓎ⌧ࡢኚ໬ࡢ㛵ಀᛶࢆ☜ㄆࡋࡓ࡜ࡇࢁ, Il1b㸦R = 0.73, p < 0.05㸧࠾ࡼࡧIl10 㑇ఏᏊ㸦R = 0.82, p < 0.01㸧࡛᭷ព࡞┦㛵ࡀࡳࡽࢀ, Il6㸦R = 0.57㸧, Ccl3㸦R = 0.65㸧࠾ࡼࡧCxcl2㑇 ఏᏊ㸦R = 0.58㸧࡛ࡣ᭷ព࡛ࡣ࡞࠿ࡗࡓࡶࡢࡢ, ┦㛵ഴྥࡀࡳࡽࢀࡓ㸦ᅗ 2-11㸧ࠋ ᐇ ᐇ㦂3 ࣭⑓⌮⤌⧊ホ౯࠾ࡼࡧ⏕໬Ꮫ᳨ᰝ  ᅄሷ໬Ⅳ⣲ࢆᢞ୚ࡋࡓࣛࢵࢺࡢ⫢⮚ࢆ⑓⌮⤌⧊Ꮫⓗ࡟ほᐹࡋࡓ࡜ࡇࢁ, ⅖⑕ᛶ⣽⬊ࡢᾐ₶ࢆకࡗࡓᑠ ⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚ࡜, ୰㛫ᇦ࡛⫢⣽⬊ࡢ⬡⫫ኚᛶࡀほᐹࡉࢀࡓ㸦ᅗ 2-12A-a㸧ࠋEEBGP ࢆᢞ୚ࡋࡓ ື≀࡛ࡣ, Control ⩌ࡢື≀࡜ྠᵝࡢኚ໬ࡀࡳࡽࢀ, ࡑࡢ⛬ᗘ࡟ᕪࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦ᅗ 2-12A-b㸧ࠋࡇࢀ ࡽࡢኚ໬࡜ྠᵝ࡟, ALT ್ࡣ Control ⩌ࡢ 211.7s112.6 U/L ࡟ᑐࡋ EEBGP ᢞ୚⩌࡛ࡣ 236.8s166.6 U/L, AST ್ࡣ Control ⩌ࡢ 560.7s185.3 U/L ࡟ᑐࡋ EEBGP ᢞ୚⩌࡛ࡣ 503.3s413.2 U/L ࡜, ୧⩌ࡢ 㛫࡟ᕪࡣࡳࡽࢀ࡞࠿ࡗࡓ㸦ᅗ2-12B㸧ࠋ ࠙⪃ᐹࠚ ᐇ㦂1 㛗ᮇࡢ⤯㣗࠶ࡿ࠸ࡣపᰤ㣴≧ែୗ࡟࠾ࡅࡿࣄࢺ࡛ࡣ, APAP ࡢ୍⯡ⓗ࡞୰ẘ㔞㸦8 g/᪥㸧ࡼࡾࡶప࠸⏝ 㔞࡛ࡶ⫢㞀ᐖࢆ⏕ࡌࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿ[40, 41]ࠋࡲࡓ, ࣐࢘ࢫ࡟࠾࠸࡚, ᢞ୚㔞ࡀྠࡌ࡛࠶ࡿ࡟ࡶ࠿ ࠿ࢃࡽࡎ, APAP ᢞ୚࡟ࡼࡿṚஸ⋡ࡸ⫢⣽⬊ቯṚࡢ⠊ᅖࡀᢞ୚᫬้࡟ࡼࡗ࡚␗࡞ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ [43]ࠋࡇࡢࡼ࠺࡞ APAP ㄏⓎᛶ⫢㞀ᐖࡢឤཷᛶࡢ㐪࠸࡟⸆≀௦ㅰ㓝⣲άᛶࡸ⫢⮚ࢢࣝࢱࢳ࢜ࣥࡀᙳ㡪ࡋ ࡚࠸ࡿࡇ࡜ࡀ♧၀ࡉࢀ࡚࠸ࡿࡓࡵ, ᐇ㦂 1 ࡛ࡣ, APAP ࡢ⸆≀ືែ࡟㛵ࢃࡿ⸆≀௦ㅰ㓝⣲άᛶ࡟ࡘ࠸࡚, ᪥ෆ࡛ࡢኚ໬࠾ࡼࡧ⤥㣵᮲௳ࡢᙳ㡪ࢆ☜ㄆࡋࡓࠋࡑࡢ⤖ᯝ, APAP ࡸ NAPQI ࡢᢪྜ࠾ࡼࡧ᤼ἥࢆಁ㐍ࡍ ࡿUgt1a6, Sult1a1࠾ࡼࡧGstm1㑇ఏᏊࡢⓎ⌧㔞ࡣ, ⮬⏤ᦤ㣵ୗ࡟࠾ࡅࡿᮅ᪉࡜ኤ᪉࡛ኚ໬ࡏࡎ, ⤯㣗 ୗࡸไ㝈⤥㣵᮲௳ୗ࡛ࡶ㢧ⴭ࡞ኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓࠋCyp2e1㑇ఏᏊࡢⓎ⌧㔞ࡣ, ⮬⏤ᦤ㣵ୗ࡟࠾ࡅࡿ ᮅ᪉࡜ኤ᪉࡛ࡣ࡯࡜ࢇ࡝ኚ໬ࡋ࡞࠿ࡗࡓࡶࡢࡢ, ⤯㣗ୗ࠾ࡼࡧไ㝈⤥㣵᮲௳ୗࡢ⮬⏤ᦤ㣵ୗ 9:00 ࡜ẚ㍑ ࡋ࡚᭷ព࡟ቑຍࡋࡓࠋCYP2E1 ࡣపศᏊ໬ྜ≀ࡢỈ㓟໬ࡸ⬺࣓ࢳࣝ໬ࢆゐ፹ࡍࡿ㓝⣲࡛࠶ࡾ, ࡑࡢ㑇ఏ ᏊⓎ⌧ࡣ, ᇶ㉁࡜࡞ࡿ໬Ꮫ≀㉁ࡢᦤྲྀࡸ኱㔞ࡢ㣧㓇࡟ࡼࡾቑຍࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[63-65]ࠋ୍᪉

23 ࡛, ⤯㣗ୗ࡛ࡶ CYP2E1 ࡀㄏᑟࡉࢀࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[47, 63, 66]ࠋ⢾ࡢ౪⤥ࡀ୙㊊ࡍࡿ࡜, ࢚ࢿ ࣝࢠ࣮୙㊊ࢆ⿵࠺ࡓࡵ࡟࢔ࢭࢳࣝCoA ࠿ࡽ࢔ࢭࢺ㓑㓟, β-ࣄࢻࣟ࢟ࢩ㓗㓟, ࢔ࢭࢺࣥ࡞࡝ࡢࢣࢺࣥయࡀྜ ᡂࡉࢀࡿࡀ[67], ࢔ࢭࢺࣥࡣ CYP2E1 ࢆㄏᑟࡋ[64, 65], ࢔ࢭࢺࣥࡢ⾑୰⃰ᗘ᫬㛫᭤⥺ୗ㠃✚࡜ CYP2E1 άᛶࡣṇࡢ┦㛵ࢆ♧ࡍࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[47]ࠋ⤯㣗ୗ[63]ࡸ 3 㐌㛫ࡢ 80%ไ㝈⤥㣵ୗ[68]࡛ࡣ⾑୰ࡢ ࢣࢺࣥయ⃰ᗘࡀቑຍࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠾ࡾ, 10 ᪥㛫ࡢ 70%ไ㝈⤥㣵࡛࠶ࡗ࡚ࡶᜏᖖⓗ࡞࢚ࢿࣝࢠ࣮ ୙㊊ࢆ⿵࠺ࡓࡵ࡟ࢣࢺࣥయ௦ㅰࡀஹ㐍ࡋ, ⤖ᯝ࡜ࡋ࡚ CYP2E1 ࡀㄏᑟࡉࢀࡓྍ⬟ᛶࡀ࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋ ࢢࣝࢱࢳ࢜ࣥࡣࢩࢫࢸ࢖ࣥ, ࢢࣝࢱ࣑ࣥ㓟, ࢢࣜࢩࣥࡢ 3 ࡘࡢ࢔࣑ࣀ㓟࠿ࡽᵓᡂࡉࢀࡿࢺࣜ࣌ࣉࢳࢻ࡛ ࠶ࡿࡀ, ࡇࢀࡽࡢ࢔࣑ࣀ㓟ࡢ࠺ࡕ, ࢩࢫࢸ࢖ࣥࡢ⏕యෆࡢ㈓ⶶ㔞ࡣከࡃ࡞࠸ୖ, ྜᡂ࡟ࡣᚲ㡲࢔࣑ࣀ㓟࡛ ࠶ࡿ࣓ࢳ࢜ࢽࣥ࡜ࢭࣜࣥࡀᚲせ࡛࠶ࡿࡇ࡜࠿ࡽ, 㣵࠿ࡽࡢ࢔࣑ࣀ㓟౪⤥ࡀῶᑡࡍࡿ࡜ࢢࣝࢱࢳ࢜ࣥࡢ⏕ ྜᡂ㔞ࡣపୗࡍࡿ[69, 70]ࠋᐇ㝿࡟ࢩࢫࢸ࢖ࣥḞஈ㣗࡛ 8 ᪥㛫㣫⫱ࡋࡓࣛࢵࢺ࡛ࡣ, ࢢࣝࢱࢳ࢜ࣥ㔞ࡣ㏻ ᖖ㣗⩌ࡢ75%ࡲ࡛ῶᑡࡋࡓ୍᪉࡛[69], ඲࡚ࡢ࢔࣑ࣀ㓟ࡢ౪⤥ࡀ࡞ࡃ࡞ࡿ 24 ᫬㛫⤯㣗ୗࡢࣛࢵࢺ࡛ࡣࢢ ࣝࢱࢳ࢜ࣥ㔞ࡣ⮬⏤ᦤ㣵ୗࡢ⣙40%ࡲ࡛పୗࡍࡿ[70]ࠋᮏ◊✲࡛ࡣ, ⤯㣗ୗ࡟ຍ࠼࡚, 10 ᪥㛫ࡢ 70%ไ㝈 ⤥㣵᮲௳ୗ࡛ࡶ⮬⏤ᦤ㣵ୗ࡜ẚ㍑ࡋ࡚᭷ព࡟పୗࡋࡓࠋQin ࡽࡣ, 1 ᪥㛫ࡢ 75%ไ㝈⤥㣵᮲௳ୗ࡛ࡣ⫢⮚ ࢢࣝࢱࢳ࢜ࣥ㔞ࡣ⮬⏤ᦤ㣵ୗ࡜ኚࢃࡽ࡞࠸࡜ሗ࿌ࡋ࡚࠸ࡿࡇ࡜࠿ࡽ[71], ᮏ◊✲ࡢࡼ࠺࡟⥅⥆ࡋࡓไ㝈 ⤥㣵ୗ࡛ࡣ㣵࠿ࡽ౪⤥ࡉࢀࡿ࢔࣑ࣀ㓟ࡀ୍ᚊ࡟ῶᑡࡍࡿࡓࡵ, ࢢࣝࢱࢳ࢜ࣥࡢ⏕ྜᡂ㔞ࡀῶᑡࡋ࡚࠸ࡿ ྍ⬟ᛶࡀ࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋຍ࠼࡚, ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࡣ᪥ෆ࡛ኚືࡍࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿ[47, 72]ࠋ ࣛࢵࢺࡣ୺࡟ᬯᮇ࡟ᦤ㣵ࡍࡿࡓࡵ, ⮬⏤ᦤ㣵ୗࡢ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࡣ, ᫂ᮇึᮇ࡟᭱ࡶ㧗ࡃ, ᫬㛫ࡢ⤒ 㐣࡟కࡗ࡚ῶᑡࡋ, ᫂ᮇᚋ༙࡛ࡣ᫂ᮇึᮇࡢ⣙ 70%ࡲ࡛ῶᑡࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࡀ[47, 72], ᮏ◊ ✲࡟࠾ࡅࡿ⮬⏤ᦤ㣵ୗࡢࢢࣝࢱࢳ࢜ࣥ㔞ࡣ᫂ᮇึᮇ㸦9:00㸧࡜᫂ᮇᚋ༙㸦16:00㸧࡛࡯࡜ࢇ࡝ኚࢃࡽ࡞࠿ ࡗࡓࠋ୍᪉, 70%ࡢไ㝈⤥㣵᮲௳ୗ࡛ࡣ, ᫂ᮇᚋ༙㸦16:00㸧ࡢࢢࣝࢱࢳ࢜ࣥ㔞ࡣ᫂ᮇึᮇ㸦9:00㸧ࡢ 44% ࡟ࡲ࡛పୗࡋ, ไ㝈⤥㣵ୗ࡛ࡣ⮬⏤ᦤ㣵ୗࡼࡾࡶ኱ࡁ࡞ኚືࡀࡳࡽࢀࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗࡓࠋ ⤯㣗ୗ࠾ࡼࡧไ㝈⤥㣵᮲௳ୗ࡛ࡣ, Cyp2e1㑇ఏᏊⓎ⌧ࡢቑຍ࡜⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࡢపୗࡀ☜ㄆࡉࢀ ࡓࡇ࡜࠿ࡽ, APAP ㄏⓎᛶࡢ⫢㞀ᐖ࡟ᙳ㡪ࡍࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋࡑࡇ࡛, ⮬⏤ᦤ㣵ୗ 9:00 ᢞ୚࡛ࡣ⫢ 㞀ᐖࢆ⏕ࡌ࡞࠸800 mg/kg య㔜ࡢ APAP ࢆ⤯㣗ୗ࠾ࡼࡧไ㝈⤥㣵᮲௳ୗࡢࣛࢵࢺ࡟ᢞ୚ࡋࡓࠋࡑࡢ⤖ᯝ, ⤯㣗ୗ࡛ࡣᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀ, ไ㝈⤥㣵᮲௳ୗ࡛ࡣ୰ᚰ㟼⬦࿘ᅖ࡟༢⣽⬊ᛶࡢ⫢⣽⬊ ቯṚࡀほᐹࡉࢀࡓࠋࡇࢀࡽࡢኚ໬ࡣ, CYP2E1 άᛶࡢቑຍ࡟కࡗࡓ NAPQI ࡢ⏕ᡂ㔞ࡢቑຍ࡜, ⫢⮚ࢢࣝ

24 ࢱࢳ࢜ࣥ㔞ࡢపୗ࡟కࡗࡓNAPQI ᤼ἥ㔞ࡢῶᑡࡢ⤖ᯝ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋຍ࠼࡚, ࡇࡢࡼ࠺࡞ప⏝㔞 ࡢ APAP ᢞ୚࡛ࡣ⫢⣽⬊ቯṚࡢ᭷↓ࡸ⛬ᗘ࡟኱ࡁ࡞ಶయᕪࡀࡳࡽࢀࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡾ, ࡑࡢཎᅉ࡜ ࡋ࡚Cyp2e1㑇ఏᏊࡢⓎ⌧㔞㸦᭱኱࡛2 ಸ㸧ࡸ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞㸦᭱኱࡛ 1.5 ಸ㸧ࡢಶయᕪࡢᙳ㡪ࡀ⪃ ࠼ࡽࢀࡓࠋ୍᪉࡛, Cyp2e1㑇ఏᏊࡢⓎ⌧㔞ࡸ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࡣ⤯㣗ୗ࡜ไ㝈⤥㣵᮲௳ୗ࡛࡯࡜ࢇ࡝ ኚࢃࡽ࡞࠿ࡗࡓ࡟ࡶ࠿࠿ࢃࡽࡎ, ⫢⣽⬊ቯṚࡢ⛬ᗘࡣ⤯㣗ୗࡢ᪉ࡀᝏ࠿ࡗࡓࡇ࡜࠿ࡽ, CYP2E1 ࡸࢢࣝࢱ ࢳ࢜ࣥ௨እࡢせᅉࡀ㛵୚ࡋ࡚࠸ࡿྍ⬟ᛶࡶ⪃࠼ࡽࢀࡓࠋᐇ㝿࡟, ࣐࢘ࢫࡸࣛࢵࢺࢆ 18㹼36 ᫬㛫⤯㣗ࡉࡏ ࡿ࡜, ᢠ㓟໬㓝⣲ࡢάᛶࡀ⮬⏤ᦤ㣵ୗࡼࡾࡶపୗࡋ, 㐣㓟໬⬡㉁ࡀቑຍࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࡀ[73, 74], 4㹼6 㐌㛫࡟ࢃࡓࡗ࡚⤥㣵㔞ࢆ⮬⏤ᦤ㣵ୗࡢ 60㹼70%࡟ไ㝈ࡋࡓࣛࢵࢺ࡛ࡣ, ࢫ࣮ࣃ࣮࢜࢟ࢩࢻࢹ࢕ ࢫ࣒ࢱ࣮ࢮ, ࢝ࢱ࣮ࣛࢮ, ࢢࣝࢱࢳ࢜ࣥ࣌ࣝ࢜࢟ࢩࢲ࣮ࢮ➼ࡢᢠ㓟໬㓝⣲ࡢάᛶࡀ⮬⏤ᦤ㣵ୗࡼࡾࡶቑ ຍࡋ, ⬡㉁㐣㓟໬ࡀᢚไࡉࢀࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[73, 75-77]ࠋࡇࢀࡽࡢሗ࿌࠿ࡽ, ᦤ㣵᮲௳㛫࡛ࡳࡽ ࢀࡓ㞀ᐖࡢ⛬ᗘࡢ㐪࠸ࡸಶయᕪࡢཎᅉ࡜ࡋ࡚, ⸆≀௦ㅰ௨እࡢせᅉࡶ㛵ಀࡋ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ 㦫ࡃ࡭ࡁࡇ࡜࡟, ⮬⏤ᦤ㣵ୗ 16:00 ࡛ࡣ, Cyp2e1㑇ఏᏊࡢⓎ⌧㔞ࡸࢢࣝࢱࢳ࢜ࣥ㔞ࡀ⮬⏤ᦤ㣵ୗ9:00 ࡜࡯࡜ࢇ࡝ኚࢃࡽ࡞࠸࡟ࡶ㛵ࢃࡽࡎ, APAP ᢞ୚࡟ࡼࡗ࡚⤯㣗ୗࡼࡾࡶከࡃࡢື≀࡛ᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ ቯṚࡀほᐹࡉࢀ, ⫢㞀ᐖ࣐࣮࣮࡛࢝࠶ࡿ ALT ࠾ࡼࡧ AST ࡀ㧗್ࢆ♧ࡋࡓࠋࡑࡢ⌮⏤ࡢ 1 ࡘ࡜ࡋ࡚, APAP ᢞ୚ᚋࡢᦤ㣵ࡢᙳ㡪ࡀ⪃࠼ࡽࢀࡓࠋࢢࣝࢱࢳ࢜ࣥࡢ⏕ྜᡂ㔞ࡣࢩࢫࢸ࢖ࣥ࠶ࡿ࠸ࡣࡑࡢ๓㥑≀㉁࡛࠶ࡿ ࣓ࢳ࢜ࢽࣥࡢ౪⤥㔞࡟౫Ꮡࡋ, ࣛࢵࢺ࡟࠾࠸࡚ APAP ᢞ୚࡟కࡗ࡚పୗࡋࡓࢢࣝࢱࢳ࢜ࣥ㔞ࢆN-࢔ࢭࢳ ࣝࢩࢫࢸ࢖ࣥࡀᅇ᚟ࡉࡏ[78], 40 ᫬㛫⤯㣗ᚋ࡟ᵝࠎ࡞⃰ᗘࡢࢩࢫࢸ࢖ࣥࢆྵࡴ㣵ࢆ୚࠼ࡓࣛࢵࢺ࡛ࡣ, ࢩ ࢫࢸ࢖ࣥ㔞౫Ꮡⓗ࡟ࢢࣝࢱࢳ࢜ࣥ㔞ࡀቑຍࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[70]ࠋࡲࡓ, APAP ᢞ୚ᚋࡢ࣓ࢳ࢜ ࢽࣥ⤒ཱྀᢞ୚ࡀ⫢㞀ᐖࢆᢚไࡍࡿ࡜ࡢሗ࿌ࡶ࠶ࡿ[79]ࠋ୍᪉, ࣐࢘ࢫ࡟࠾࠸࡚, ࢢࣝࢱࢳ࢜ࣥࡀ༑ศ࡟ṧ ࡗ࡚࠸ࡿ࡟ࡶ࠿࠿ࢃࡽࡎ NAPQI ࡢࢩࢫࢸ࢖ࣥ௜ຍయࡀ᳨ฟࡉࢀࡓ [80]ࠋࡇࡢࡇ࡜ࡣ NAPQI ࡀࢢࣝࢱ ࢳ࢜ࣥᏑᅾୗ࡛ࡶ㠀≉␗ⓗ࡟⣽⬊ෆࡢࢳ࣮࢜ࣝ࡜⤖ྜࡍࡿྍ⬟ᛶࢆ♧၀ࡋ࡚࠾ࡾ, 㣵⏤᮶ࡢࢩࢫࢸ࢖ࣥ ࡸ࣓ࢳ࢜ࢽࣥࡀ⫢⮚࡟⛣⾜ࡋ, ┤᥋ NAPQI ࡜⤖ྜࡋ࡚⫢㞀ᐖࢆᢚไࡋࡓྍ⬟ᛶࡶ࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋ ⸆≀௦ㅰ㓝⣲ࡀᦤ㣵᮲௳ࡢᙳ㡪ࢆཷࡅࡿࡇ࡜ࡀ᫂ࡽ࠿࡛࠶ࡗࡓࡓࡵ, APAP ࡜ྠࡌࡃẘᛶⓎ⌧࡟ CYP2E1 ࡀ㛵ࢃࡿᅄሷ໬Ⅳ⣲ㄏⓎᛶࡢ⫢㞀ᐖ࡟ࡘ࠸࡚, ⮬⏤ᦤ㣵ୗ࡜⤯㣗ୗ࡛㞀ᐖࡢ⛬ᗘࢆẚ㍑ࡋࡓࠋ ࡑࡢ⤖ᯝ, ⤯㣗ୗ࡛ࡣ⮬⏤ᦤ㣵ୗࡼࡾࡶᗈ࠸⠊ᅖ࡛⫢⣽⬊ቯṚࡀㄆࡵࡽࢀ, ᪤▱ࡢሗ࿌࡜୍⮴ࡋࡓ[47, 71]ࠋࡇࢀࡽࡢሗ࿌࡛ࡶ, ⤯㣗࡟ࡼࡾCyp2e1㑇ఏᏊࡢⓎ⌧㔞ࡢቑຍࡸCYP2E1 άᛶࡢቑຍࡀ♧ࡉࢀ࡚࠸

25

ࡿࡇ࡜࠿ࡽ, ⤯㣗࡟ࡼࡿᅄሷ໬Ⅳ⣲ࡢ⫢ẘᛶࡢቑᙉࡣ, CYP2E1 ࡢㄏᑟࡀ཯ᫎࡉࢀࡓࡶࡢ࡜⪃࠼ࡽࢀࡓࠋ ᐇ

ᐇ㦂2

EEBGP ࢆ APAP ࡢᢞ୚࡟ඛ❧ࡗ࡚ 7 ᪥㛫ᢞ୚ࡍࡿࡇ࡜࡟ࡼࡾ, APAP ࡟ࡼࡿ⫢⣽⬊ቯṚࡢ⠊ᅖࡀ㍍ῶ ࡍࡿࡇ࡜ࡀ☜ㄆࡉࢀࡓࠋAPAP ࡀ⫢⣽⬊ቯṚࢆㄏᑟࡍࡿ࣓࢝ࢽࢬ࣒࠿ࡽ, EEBGP ࡀ, (i) άᛶ㓟⣲✀ࡢᾘ ཤ, (ii) ⅖⑕཯ᛂࡢㄪᩚ, (iii) APAP ࠾ࡼࡧ NAPQI ࡢ௦ㅰไᚚ࠾ࡼࡧ᤼ἥಁ㐍࡟ᙳ㡪ࡋ࡚࠸ࡿྍ⬟ᛶࡀ⪃ ࠼ࡽࢀࡓࠋ

EEBGP ࡣ, RAW294.6 ⣽⬊࡟࠾࠸࡚ᢠ㓟໬㓝⣲ࡢ mRNA 㔞࠾ࡼࡧࢱࣥࣃࢡ㔞ࢆቑຍࡉࡏ, άᛶ㓟⣲ ✀ࡢ⏘⏕ࢆపୗࡉࡏࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[11]ࠋࡋ࠿ࡋ࡞ࡀࡽ, ᮏᐇ㦂࡛ࡣ, ᢠ㓟໬㓝⣲㑇ఏᏊࡢⓎ⌧ 㔞࡟ࡣኚ໬ࡀࡳࡽࢀࡎ, ᢠ㓟໬㓝⣲ࡢㄏᑟ࡟ࡼࡗ࡚άᛶ㓟⣲✀ࢆᾘཤࡍࡿྍ⬟ᛶࡣప࠸࡜⪃࠼ࡽࢀࡓࠋ୍ ᪉, ᮏᐇ㦂࡛ࡣ, ⫢⮚࡟࠾࠸࡚⅖⑕཯ᛂࢆಁ㐍ࡍࡿIl1b, Il6, Cxcl2࡜࠸ࡗࡓ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥࡸࢣࣔ ࢝࢖ࣥ㑇ఏᏊࡢⓎ⌧㔞ࡀEEBGP ᢞ୚࡟ࡼࡾపୗࡋࡓࠋຍ࠼࡚, ࡇࢀࡽࡢ㑇ఏᏊࡢⓎ⌧㔞, ≉࡟Il1b㑇 ఏᏊ࡜⫢⣽⬊ቯṚࡢ⠊ᅖ࡟┦㛵ࡀࡳࡽࢀࡓࡇ࡜࠿ࡽ, EEBGP ᢞ୚ࡣ⅖⑕཯ᛂࢆಁ㐍ࡍࡿ㑇ఏᏊࡢⓎ⌧ୖ ᪼ࢆᢚ࠼, ⅖⑕཯ᛂࢆㄪᩚࡋࡓྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋᢠ IL-1 ᢠయࡸ IL-1 ཷᐜయḞᦆ࣐࢘ࢫࢆ⏝࠸ࡓሗ࿌ ࠿ࡽ, APAP ࡟ࡼࡿ⫢㞀ᐖࡢ㐍ᒎ࡟ࡣ IL-1α ࠾ࡼࡧ IL-1β ࡢάᛶ໬ࡀᐦ᥋࡟㛵ࢃࡾ[81, 82], ⫢㞀ᐖࢆ㉳ࡇ ࡋࡓ࣐࢘ࢫ࡟࠾࠸࡚, IL-1β ࡢᢚไࡣࡑࡢ௚ࡢࢧ࢖ࢺ࢝࢖ࣥ⏘⏕ࢆᢚไࡋ, ⅖⑕཯ᛂ࡟క࠺⤌⧊㞀ᐖࡢ㍍ ῶ࡟㔜せ࡞ᙺ๭ࢆࡶࡘࡇ࡜ࡀᣦ᦬ࡉࢀ࡚࠸ࡿ[83]ࠋ୍᪉, ⫢⤌⧊ࡢയᐖࡢక࠸⅖⑕཯ᛂࡀ⏕ࡌࡿ࡜, ⅖⑕ ᛶࢧ࢖ࢺ࢝࢖ࣥ࡜࡜ࡶ࡟, ⅖⑕཯ᛂࢆ㈇࡟ࣇ࢕࣮ࢻࣂࢵࢡไᚚࡍࡿᢠ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥࡀㄏᑟࡉࢀࡿ [64, 65]ࠋ࣐࢘ࢫ࡬ࡢ APAP ᢞ୚࡛ࡶ௦⾲ⓗ࡞ᢠ⅖⑕ᛶࢧ࢖ࢺ࢝࢖࡛ࣥ࠶ࡿIl10㑇ఏᏊࡢⓎ⌧㔞ࡀቑຍ ࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[86, 87]ࠋKannno ࡽࡣ APAP ࡟ࡼࡿ⫢㞀ᐖ࡟కࡗ࡚⾑୰ࡢIl10㑇ఏᏊࡢ mRNA ࡀቑຍࡍࡿࡇ࡜ࢆሗ࿌ࡋ࡚࠾ࡾ[87], EEBGP ᢞ୚⩌࡛ࡳࡽࢀࡓIl10㑇ఏᏊࡢⓎ⌧㔞ࡢపୗࡣ,

EEBGP ࡀ┤᥋Il10㑇ఏᏊࡢⓎ⌧࡟ᙳ㡪ࡋࡓ࡜࠸࠺ࡼࡾࡶ⫢⣽⬊ቯṚࡢ㍍ῶ࡟కࡗࡓኚ໬࡛࠶ࡿྍ⬟ᛶ ࡀ⪃࠼ࡽࢀࡓࠋᐇ㝿࡟, ᮏᐇ㦂࡛ࡣ, Il10 㑇ఏᏊࡢⓎ⌧㔞࡜⫢⣽⬊ቯṚࡢ⠊ᅖ࡟┦㛵ࡀࡳࡽࢀࡓࠋ ᮏヨ㦂࡛ࡣ, ᢠ㓟໬㓝⣲㑇ఏᏊ࡟ࡣኚ໬ࡀࡳࡽࢀࡎ, ࢧ࢖ࢺ࢝࢖ࣥཬࡧࢣࣔ࢝࢖ࣥ㑇ఏᏊࡢኚ໬ࡢࡳ ࡀ☜ㄆࡉࢀ, ࡇࢀ࡟ࡣࢧࣥࣉࣜࣥࢢࡢࢱ࢖࣒࣏࢖ࣥࢺࡀ㛵ಀࡋ࡚࠸ࡿ࠿ࡶࡋࢀ࡞࠸࡜⪃࠼ࡽࢀࡓࠋ࣐࢘ࢫ ࡟APAP ᢞ୚ࡍࡿ࡜ 1 ᫬㛫௨ෆ࡟ࢢࣝࢱࢳ࢜ࣥࡀᯤῬࡋ[88, 89], ࡑࡢᚋࡍࡄ࡟άᛶ㓟⣲✀ࡀ⏘⏕ࡉࢀࡿ ࡓࡵ[89-91], ᢠ㓟໬㓝⣲ࡢㄏᑟࡣᢞ୚ 24 ᫬㛫ᚋࡼࡾࡶ᪩࠸ࢱ࢖࣑ࣥࢢ࡛㉳ࡇࡗ࡚࠸ࡿྍ⬟ᛶࡀ࠶ࡿࠋ୍

26

᪉࡛, ዲ୰⌫➼ࡢ⅖⑕ᛶ⣽⬊ࡢᾐ₶ࡣᢞ୚ 24 ᫬㛫ᚋ࡛ࡶ⥅⥆ࡋ࡚࠸ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[91]ࠋ BGP ࡜ࡣ㉳※᳜≀ࡸྵ᭷ᡂศࡀ␗࡞ࡿ, ࣏ࣉࣛࢱ࢖ࣉࣉ࣏ࣟࣜࢫࡢ࢚ࢱࣀ࣮ࣝᢳฟ≀ࢆ࣐࢘ࢫࡸࣛࢵ ࢺ࡟7 ᪥㛫㐃⥆ࡋ࡚⤒ཱྀᢞ୚ࡍࡿ࡜, CYP2E1 άᛶࡀపୗࡋ, APAP ࡸ NAPQI ࡢ᤼ἥ࡟㛵ࢃࡿ

Sulfotransferase άᛶ࠾ࡼࡧ Glutathione-S-transferase άᛶࡀቑຍࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ[14], ࣏ࣉࣛ⣔

ࣉ࣏ࣟࣜࢫࡢ≉ᚩᡂศ࡛࠶ࡿCAPE ࢆ 3 ᪥㛫㐃⥆ࡋ࡚⤒ཱྀᢞ୚ࡍࡿࡇ࡜࡟ࡼࡾ CYP2E1 ࢱࣥࣃࢡ㔞࠾ࡼ

ࡧCYP2E1 άᛶࡀపୗࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[92]ࠋᮏᐇ㦂࡛⏝࠸ࡓ EEBGP ࡣ CYP2E1 άᛶ࡟ᑐࡍ ࡿሗ࿌ࡢ࠶ࡗࡓCAPE ࢆྵࢇ࡛࠸࡞࠸ࡀ[11, 59], EEBGP ࡀ⸆≀௦ㅰ㓝⣲࡟ᙳ㡪ࡋࡓྍ⬟ᛶࡣ᏶඲࡟ࡣ ྰᐃ࡛ࡁ࡞࠿ࡗࡓࠋ ᐇ ᐇ㦂3  EEBGP ࢆᅄሷ໬Ⅳ⣲ࡢᢞ୚࡟ඛ❧ࡗ࡚ 7 ᪥㛫ᢞ୚ࡋࡓࡀ, ᅄሷ໬Ⅳ⣲࡟ࡼࡿ⫢⣽⬊ቯṚࡢ⠊ᅖࡸ⬡⫫ ኚᛶࡢ⛬ᗘࡣControl ⩌࡜ኚࢃࡽ࡞࠿ࡗࡓࠋᅄሷ໬Ⅳ⣲࡟ࡼࡿ⫢㞀ᐖࡣ, ࣭CCl3ࡸ࣭OOCCl3ࡢࡼ࠺࡞ࣛ ࢪ࢝ࣝ࡟ࡼࡿ⬡㉁㐣㓟໬ࢆཎᅉ࡜ࡍࡿࡓࡵ[44, 45], ࣅࢱ࣑ࣥ E [52]ࡸN-࢔ࢭࢳࣝࢩࢫࢸ࢖ࣥ[53]➼ࡢ┤ ᥋ࣛࢪ࢝ࣝࢆᾘཤࡍࡿ≀㉁࡛ࡢಖㆤຠᯝࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋඛ⾜◊✲࡛ࡣ, EEBGP ࡸ࢔ࣝࢸࣆࣜࣥ C ࡀ, άᛶ㓟⣲✀[58]ࡸ 2, 2-ࢪࣇ࢙ࢽࣝ-1-ࣆࢡࣜࣝࣄࢻࣛࢪࣝ㸦DPPH㸧ࣛࢪ࢝ࣝ[59, 93]ࢆ㠀㓝⣲ⓗ࡟┤᥋ᾘ ཤࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋᡃࠎࡣ, ࣛࢵࢺ࡬ࡢ 500mg/kg య㔜ࡢ EEBGP ࡢ༢ᅇᙉไ⤒ཱྀᢞ୚࡟ࡼࡾ, EEBGP ୰ࡢ୺せᡂศ࡛࠶ࡿp-ࢡ࣐ࣝ㓟, ࢻࣝࣃࢽࣥ࠾ࡼࡧ࢔ࣝࢸࣆࣜࣥ C ࡢ⾑୰⃰ᗘࡣᢞ୚ᚋ 10㹼20 ศ࡛ࡑࢀࡒࢀ᭱㧗⾑୰⃰ᗘ࡟㐩ࡋ, ᢞ୚ 1 ᫬㛫ᚋ࡟ࡣ p-ࢡ࣐ࣝ㓟ࡣࡑࡢ 1/10 ࡲ࡛పୗࡋ, ࢻࣝࣃࢽࣥ࠾ ࡼࡧ࢔ࣝࢸࣆࣜࣥC ࡣ⾑୰࠿ࡽ࡯࡜ࢇ࡝ᾘኻࡍࡿࡇ࡜ࢆ☜ㄆࡋ࡚࠾ࡾ㸦Data not shown㸧, ΰ㣵ᢞ୚࡛ ࡣ, ⫢⤌⧊୰ࡢ⃰ᗘࡀ⫢㞀ᐖࢆᢚไࡍࡿࡢ࡟༑ศ࡞⃰ᗘࡲ࡛ୖ᪼ࡋ࡞࠿ࡗࡓྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋᅄሷ໬ Ⅳ⣲࡟ࡼࡿ⫢㞀ᐖࡣ, ࢡࢵࣃ࣮⣽⬊ࡢḞᦆ[56, 57]ࡸ TNF-α ࡢ㜼ᐖ[94]࡟ࡼࡾῶᙅࡍࡿ୍᪉࡛, IL-1 ࡢࢩ ࢢࢼࣝఏ㐩ࢆ㜼ᐖࡍࡿ࡜, ᅄሷ໬Ⅳ⣲ᢞ୚ 24 ᫬㛫ᚋࡢ⫢⣽⬊ቯṚ࡟ࡣ࡯࡜ࢇ࡝ᙳ㡪ࡏࡎ, ᢞ୚ 3 ᪥ᚋ࡟ ࠾࠸࡚⫢⣽⬊ቯṚࡢ⠊ᅖࡀపୗࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[95]ࠋࡲࡓ, ᅄሷ໬Ⅳ⣲ࡢᢞ୚࡟ࡼࡾ⫢⮚࡛㐣 㓟໬ࢪ࢔ࢩࣝࢢࣜࢭ࣮ࣟࣝࡀ⏕ࡌ, ࣉࣟࢸ࢖ࣥ࢟ࢼ࣮ࢮ C ࢆ௓ࡋ࡚ Nuclear factor-κB㸦NF-κB㸧ࢆάᛶ ໬ࡋ, ⫢⣽⬊ቯṚࢆㄏᑟࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[96]ࠋࡇࢀࡽࡢሗ࿌࠿ࡽ, ⤌⧊㞀ᐖ࡜⅖⑕཯ᛂ࡜ࡢ㛵 ಀࡣᅄሷ໬Ⅳ⣲࡜APAP ࡛ࡣ␗࡞ࡿࡇ࡜ࡀண᝿ࡉࢀ, ୧⪅ࡢ㛫࡛ EEBGP ࡢస⏝ࡀ␗࡞ࡗࡓ୍ᅉ࡛࠶ࡿ ྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ

27 ࠙ᑠᣓࠚ ࣄࢺ࡟࠾࠸࡚, ୍⯡ⓗ࡞୰ẘ㔞ࡼࡾࡶᑡ࡞࠸㔞࡟ࡶ࠿࠿ࢃࡽࡎ, APAP ࡢᦤྲྀ࡟ࡼࡾ⫢㞀ᐖࢆ⏕ࡌࡿሙ ྜࡀ࠶ࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠾ࡾ, ື≀ヨ㦂࡟࡚ APAP ࡢ௦ㅰ࡟㛵ࢃࡿ⸆≀௦ㅰ㓝⣲ࡸ⫢⮚ࢢࣝࢱࢳ࢜ࣥࡢ 㛵୚ࡀ♧၀ࡉࢀ࡚࠸ࡿࠋᐇ㦂1 ࡛ࡣ, ࡇࡢࡼ࠺࡞ప⏝㔞ࡢ APAP ᦤྲྀ᫬࡟⏕ࡌࡿ⫢㞀ᐖࢆᶍࡋࡓࣔࢹࣝ ࢆస〇ࡍࡿ┠ⓗ࡛, 1 ᪥ࡢ␗࡞ࡿ᫬㛫࡟࠾ࡅࡿ⸆≀௦ㅰ㓝⣲ࡢ㑇ఏᏊⓎ⌧㔞࠾ࡼࡧ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࢆ ☜ㄆࡋ, ຍ࠼࡚ᦤ㣵᮲௳ࡢᙳ㡪ࢆ☜ㄆࡋࡓࠋࡑࡢ⤖ᯝ, ࡍ࡛࡟ሗ࿌ࡢ࠶ࡿ⤯㣗ୗ࡟ຍ࠼࡚, 70%ࡢไ㝈⤥ 㣵ࢆ10 ᪥㛫⥅⥆ࡋࡓ᮲௳ୗ࡛ࡶ, ⮬⏤ᦤ㣵ୗ࡜ẚ㍑ࡋ࡚Cyp2e1㑇ఏᏊࡢⓎ⌧㔞ࡀቑຍࡋ, ࢢࣝࢱࢳ࢜ ࣥ㔞ࡀపୗࡋࡓࠋ୍᪉, ⮬⏤ᦤ㣵ୗ࡟࠾ࡅࡿ 9:00 ࡜ 16:00 ࡢẚ㍑࡛ࡣ, Cyp2e1㑇ఏᏊࡢⓎ⌧㔞࠾ࡼࡧࢢ ࣝࢱࢳ࢜ࣥ㔞࡟㢧ⴭ࡞ኚ໬ࡣࡳࡽࢀ࡞࠿ࡗࡓࠋྛ᮲௳ୗ࡟࠾࠸࡚, ㏻ᖖࡢ⮬⏤ᦤ㣵ୗࡢᢞ୚㸦9:00 ᢞ୚㸧 ࡛ࡣ⫢⣽⬊ቯṚࡀࡳࡽࢀ࡞࠸800 mg/kg య㔜ࡢ APAP ࢆᢞ୚ࡋࡓ࡜ࡇࢁ, ⤯㣗ୗ࠾ࡼࡧไ㝈⤥㣵᮲௳ୗ ࡛⫢⣽⬊ቯṚࢆ⏕ࡌ, ࡇࢀࡣ CYP2E1 ࠾ࡼࡧࢢࣝࢱࢳ࢜ࣥࡢኚ໬࡟కࡗ࡚ APAP ࡢ௦ㅰࡀኚ໬ࡋࡓࡓࡵ ࡜⪃࠼ࡽࢀࡓࠋ୍᪉, ⮬⏤ᦤ㣵ୗ 16:00 ᢞ୚࡛ࡣ, ⤯㣗ୗࡸไ㝈⤥㣵ୗࡼࡾࡶከࡃࡢື≀࡛⫢⣽⬊ቯṚࡀ ほᐹࡉࢀ, Cyp2e1 㑇ఏᏊࡸࢢࣝࢱࢳ࢜ࣥ㔞ࡢኚ໬ࡼࡾࡶ, APAP ᢞ୚ᚋࡢᦤ㣵ࡢ᭷↓ࡀ⫢⣽⬊ቯṚࡢ⛬ ᗘ࡟ࡼࡾ኱ࡁࡃᙳ㡪ࡍࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋຍ࠼࡚, ࡇࡢࡼ࠺࡞ప⏝㔞ࡢ APAP ᢞ୚࡛ࡣ⫢⣽⬊ቯṚ ࡢ᭷↓ࡸ⛬ᗘ࡟኱ࡁ࡞ಶయᕪࡀࡳࡽࢀࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡾ, ࡑࡢཎᅉ࡜ࡋ࡚ Cyp2e1 㑇ఏᏊࡢⓎ⌧㔞 ࡸ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞ࡢಶయᕪࡢᙳ㡪ࡀ⪃࠼ࡽࢀࡓࠋᮏᐇ㦂࡛ᐇ᪋ࡋࡓྛᢞ୚᮲௳ࡢ࠺ࡕ, APAP ࡢᢞ୚ ᮲௳࡜ࡋ࡚᭱ࡶᙉ࠸㞀ᐖࡀࡳࡽࢀࡓ⮬⏤ᦤ㣵ୗ16:00 ᢞ୚ࢆᐇ㦂 2 ࡟⏝࠸ࡿࡇ࡜࡜ࡋࡓࠋࡲࡓ, APAP ࡜ ࡣ␗࡞ࡿ⫢㞀ᐖࣔࢹࣝ࡜ࡋ࡚, ᅄሷ໬Ⅳ⣲ࢆ⮬⏤ᦤ㣵ୗ࠾ࡼࡧ⤯㣗ୗ࡛ᢞ୚ࡋࡓ࡜ࡇࢁ, ⤯㣗ୗ࡛ࡼࡾ ᙉ࠸㞀ᐖࡀほᐹࡉࢀࡓࡀ, ࡇࢀࡣ⤯㣗࡟ࡼࡾㄏᑟࡉࢀࡓ CYP2E1 ࡟㉳ᅉࡋࡓࡶࡢ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡓࠋ  ᐇ㦂2 ࡛, APAP ㄏⓎᛶ⫢㞀ᐖ࡟ᑐࡍࡿ EEBGP ࡢᢚไຠᯝࢆ᳨ウࡋࡓ࡜ࡇࢁ, 0.3% EEBGP ࢆ APAP

ᢞ୚๓࡟7 ᪥㛫, ΰ㣵ᢞ୚ࡍࡿࡇ࡜࡛⫢⣽⬊ቯṚࡢ⠊ᅖࡀ᭷ព࡟పୗࡋࡓࠋಖㆤຠᯝࡢ࣓࢝ࢽࢬ࣒ࢆ☜ㄆ ࡍࡿࡓࡵ, ᢠ㓟໬㓝⣲㑇ఏᏊ࠾ࡼࡧ⅖⑕㛵㐃㑇ఏᏊࡢⓎ⌧࡬ࡢᙳ㡪ࢆ☜ㄆࡋࡓ࡜ࡇࢁ, EEBGP ࡢᢞ୚ࡣ ᢠ㓟໬㓝⣲㑇ఏᏊࡢⓎ⌧࡟ࡣ࡯࡜ࢇ࡝ᙳ㡪ࢆ୚࠼ࡎ, ࢧ࢖ࢺ࢝࢖ࣥࡸࢣࣔ࢝࢖ࣥࡢࡼ࠺࡞⅖⑕㛵㐃ᅉᏊ ࡢ㑇ఏᏊⓎ⌧ࢆపୗࡉࡏࡓࠋࡲࡓ, Il1b㑇ఏᏊࡸIl10㑇ఏᏊࡢⓎ⌧పୗ࡜⫢⣽⬊ቯṚࡢ㍍ῶ࡟ࡣ┦㛵ࡀ ࡳࡽࢀࡓࡇ࡜࠿ࡽ, EEBGP ࡢ⫢㞀ᐖᢚไ࣓࢝ࢽࢬ࣒ࡢ 1 ࡘ࡜ࡋ࡚, ⅖⑕཯ᛂ࡟ᙳ㡪ࡋ࡚࠸ࡿྍ⬟ᛶࡀ⪃ ࠼ࡽࢀࡓࠋ

28

 ᐇ㦂3 ࡛, ᅄሷ໬Ⅳ⣲ㄏⓎᛶ⫢㞀ᐖ࡟ᑐࡍࡿ EEBGP ࡢᢚไຠᯝࢆ᳨ウࡋࡓ࡜ࡇࢁ, 1.2% EEBGP ࡢ 7 ᪥㛫ΰ㣵ᢞ୚ࡣ, ⫢⣽⬊ቯṚ࠾ࡼࡧ⫢⣽⬊ࡢ⬡⫫ኚᛶ࡟ᙳ㡪ࡋ࡞࠿ࡗࡓࠋᅄሷ໬Ⅳ⣲࡟ࡼࡿ⫢㞀ᐖ࡛ࡣ, CYP2E1 ௦ㅰ≀࡛࠶ࡿࣛࢪ࢝ࣝࡢᾘཤࡀ㔜せ࡛࠶ࡾ, EEBGP ࡀࣛࢪ࢝ࣝᾘཤస⏝ࢆ᭷ࡍࡿࡇ࡜ࡀሗ࿌ࡉ ࢀ࡚࠸ࡿࡶࡢࡢ, ௒ᅇࡢᐇ㦂᮲௳࡛ࡣ㞀ᐖࢆᢚไࡍࡿ࡟ࡣ୙༑ศ࡛࠶ࡗࡓྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ

29 ࠙ᅗ⾲ࠚ

⾲2-1. ࣉࣛ࢖࣐࣮ࣜࢫࢺ

F: Forward primer, R: Reverse primer

F 5'-AACATGATCTTCATTGGAGGGAC-3' R 5'-CCATCGCTTTCTTCTCTGGAATC-3' F 5'-CCACGGCTCCTTAAGACACA-3' R 5'-AGACAACCACATCCTTTGCATTT-3' F 5'-CTGAGACCACCAGCACAACT-3' R 5'-GCCCAATAACCCTGTCAATTTCT-3' F 5'-ATGATACTGGGATACTGGAACGT-3' R 5'-CACTGGCTTCTGTCATAGTCG-3' F 5'-GACATGCCGCCTGGAGAA-3' R 5'-AGCCCAGGATGCCCTTTAGT-3' Sult1a1 NM_031834.1 Gapdh NM_017008.4 Gstm1 NM_017014.1 Cyp2e1 NM_031543.1

Gene name F or R Sequence Accession no.

30 ⾲2-2. ࣉࣛ࢖࣐࣮ࣜࢫࢺ

F: Forward primer, R: Reverse primer

Gene name F or R Primer sequence Acccession

No. F CAGGATTAACTGAAGGCGAGCAT R GCTGGACCGCCATGTTTCTT F GCTGGCTTGGCTTCAATAAGG R GTGCTCCCACACATCAATCC F AGCCAGAAGAGAAACCCACAAA R CCATTCATGTGCCGATGTCC F CGGGACTACACCGAAATGAATG R TCACCTCGCACTTCTCAAACAA F GTGCATCGTCACCAACGTG R CGGCTGCAAACTCCTTGATT F TCAGATCCCAAGCGCACCAT R AGCGGCCAACAGGAAGATCA F GTGACCACGAGGAAGACGAAA R CATCTCATCGCAGCCAATCC Cytokines F TGAGGACCCAAGCACCTTCT R TGGGAACATCACACACTAGCA F GTCAACTCCATCTGCCCTTCA R TGTCAACAACATCAGTCCCAAGA F AGTGGAGCAGGTGAAGAATGA R CAGTAGATGCCGGGTGGTT F CCAGCTTCATACCTCCCTACT R AGGATCTTGGAACGGAGAAGA F GGTTCCGTCCCTCTCATACAC R TCCACATCTCGGATCATGCTTT F CTCTCTTCCTCCACCACTATGC R GTAGTTCTCCAGCCGACTCATT F ACAAGCGCACCCTCTGTT R GAATTTGCCGTCCATAGGAGAA F GCGCCCAGACAGAAGTCATA R CGAGGCACATCAGGTACGAT F TCAGGCTTGGGTCTTGTTAGC R ATGTCTGTGACTTTGTGCTTCTG Gpx4 NM_012854.2 Il10

Antioxidative enzyme genes

Inflammatory-related genes Il6 NM_012589.2 NM_031530.1 Tnf NM_012675.3 Chemokines Ccl2 Nos2 NM_012611.3 Ccl3 Cxcl2 NM_053647.1 NM_013025.2

Production of inflammatory mediators

Il23a NM_130410.2 Gsr NM_053906.2 Il1b NM_031512.2 Gpx1 NM_030826.4 Prdx1 NM_057114.1 NM_017165.3 Sod2 NM_017051.2 Cat NM_012520.2 Sod1 NM_017050.1

31

⾲2-3. ␗࡞ࡿᦤ㣵᮲௳ୗ࡟࠾ࡅࡿ࢔ࢭࢺ࢔࣑ࣀࣇ࢙ࣥᢞ୚㸦800 mg/kg య㔜㸧24 ᫬㛫ᚋࡢ⑓⌮⤌⧊Ꮫ᳨ ᰝࡢ⤖ᯝ, ⾑୰ ALT ್࠾ࡼࡧ AST ್

+: ⫢⣽⬊ቯṚ࠶ࡿ࠸ࡣ⅖⑕ᛶ⣽⬊ᾐ₶࠶ࡾ, -: ⫢⣽⬊ቯṚ࠶ࡿ࠸ࡣ⅖⑕ᛶ⣽⬊ᾐ₶࡞ࡋ ⮬⏤ᦤ㣵ୗࡢᖹᆒ್sᶆ‽೫ᕪ ALT: 51.0s9.2 U/L, AST: 84.7s8.0 U/L

Inflammatory ALT AST

Centrilobular Single cell cell infiltration (U/L) (U/L)

- - - 41 84 - - - 53 77 - - - 59 93 + - + 192 973 - + + 51 130 + - + 83 261 + - + 393 2896 + - + 722 5080 + - + 92 283 + - + 82 380 - + + 49 115 - + + 53 125 - - - 40 79 - + + 51 100 - - - 37 61 - + + 48 83 - + + 59 102 - - - 39 59 Restricted feeding 16:00 Ad libitum feeding 16:00 Hepatocellular necrosis 9:00 16 hours fasting 9:00 Feeding condition Administered time

32 ᅗ2-1. APAP ࡢẘᛶⓎ⌧࣓࢝ࢽࢬ࣒

APAP ࡢ኱㒊ศࡣࢢࣝࢡࣟࣥ㓟ᢪྜ࠾ࡼࡧ◲㓟ᢪྜࢆཷࡅ࡚ᒀ୰࡬᤼ἥࡉࢀࡿࡀ, ୍㒊ࡣ CYP2E1 ࡟ࡼࡾ NAPQI ࡟௦ㅰࡉࢀ, ㏿ࡸ࠿࡟ࢢࣝࢱࢳ࢜ࣥᢪྜࢆཷࡅ࡚ᒀ୰࡬࡜᤼ἥࡉࢀࡿࠋࡋ࠿ࡋ࡞ࡀࡽ, ୍ᗘ࡟኱㔞ࡢ APAP ࢆᦤྲྀࡍࡿ࡜, ᤼ἥࡉࢀ࡞࠿ࡗࡓ NAPQI ࡀ⣽⬊ᶵ⬟ࢆ㞀ᐖࡋ, ⏘⏕ࡋࡓάᛶ㓟⣲✀㸦ROS㸧࡟ࡼࡗ࡚ ⫢⣽⬊ࡀቯṚࡍࡿࠋቯṚࡋࡓ⣽⬊ࡣ, ⅖⑕ᛶ⣽⬊࡟ࡼࡾ㝖ཤࡉࢀࡿࡀ, 㐣๫࡞⅖⑕཯ᛂࡣṇᖖ࡞⫢⣽⬊ࡲ࡛◚ ቯࡋ, ⫢⣽⬊ቯṚࡢ⠊ᅖࢆᣑ኱ࡉࡏࡿࠋ

33 ᅗ2-2. ᅄሷ໬Ⅳ⣲ࡢẘᛶⓎ⌧࣓࢝ࢽࢬ࣒ ᅄሷ໬Ⅳ⣲ࡀCYP2E1 ࡟ࡼࡗ࡚௦ㅰࡉࢀࡿࡇ࡜࡛, ࣭CCl3ࡸ࣭OOCCl3࡞࡝ࡢࣛࢪ࢝ࣝࡀ⏕ࡌࡿࠋࡇࢀࡽ ࡢࣛࢪ࢝ࣝࡀࢱࣥࣃࢡࡸ⬡㉁➼ࡢ⏕య㧗ศᏊࢆ㓟໬ⓗ࡟യᐖࡋ, ⫢⣽⬊ࢆቯṚࡉࡏࡿࠋࡲࡓ, ࢔࣏࣏ࣜࢱࣥࣃ ࢡ㉁ࡢྜᡂࡀ㜼ᐖࡉࢀࡿࡇ࡜࡛, ࢺࣜࢢࣜࢭࣜࢻࡢ㍺㏦ࡀࡾ, ⫢⮚୰࡟⬡⫫⁲ࡀ⵳✚ࡍࡿࠋࣛࢪ࢝ࣝࡢ୍㒊 ࡣ࣍ࢫࢤࣥ㸦OCCl2㸧ࢆ⤒࡚, ஧㓟໬Ⅳ⣲㸦CO2㸧࡜ሷ㓟㸦HCl㸧࡟ศゎࡉࢀࡿࠋ ᅗ2-3. ไ㝈⤥㣵ୗ࡟࠾ࡅࡿయ㔜᥎⛣ ไ㝈⤥㣵ୗ㸦⮬⏤ᦤ㣵ୗࡢ70%; 14 g/᪥㸧࡛࠶ࡗ࡚ࡶయ㔜ࡢపୗࡣࡳࡽࢀ࡞࠿ࡗࡓࠋ

34

ᅗ2-4. ␗࡞ࡿᦤ㣵᮲௳ୗ࡟࠾ࡅࡿ UDP-glucuronosyltransferase 1A6 (Ugt1a6), sulfotransferase 1A1 (Sult1a1), cytochrome P450 2E1 (Cyp2e1)࠾ࡼࡧ glutathione-S-transferase M1 (Gstm1) 㑇ఏᏊࡢ mRNA 㔞

ྛ㑇ఏᏊࡢⓎ⌧㔞ࡣ Gapdh ࡟ᑐࡍࡿ┦ᑐ್࡜ࡋࡓࠋྛⅬࡣྛಶయࡢ㑇ఏᏊⓎ⌧㔞ࢆ♧ࡋ, ᶓᲬࡣྛ⩌ࡢᖹ ᆒ㸦୰ኸ㸧࠾ࡼࡧᶆ‽೫ᕪ㸦ୖୗ㸧ࢆ, ᣓᘼࡣ᥇ᮦ᫬㛫ࢆ♧ࡍࠋྛ㑇ఏᏊࡢⓎ⌧㔞ࡣ Gapdh ࡢⓎ⌧㔞࡛⿵ṇ ࡋࡓࠋALF: ⮬⏤ᦤ㣵ୗ, Fasting: 16 ᫬㛫⤯㣗ୗ, RF: ไ㝈⤥㣵ୗ㸦⮬⏤ᦤ㣵ࡢ 70%㸧ࠋ **: p < 0.01 vs ALF ⩌ (9:00)㸦Steel ࡢከ㔜ẚ㍑᳨ᐃ㸧ࠋ

35 ᅗ2-5. ␗࡞ࡿᦤ㣵᮲௳ୗ࡟࠾ࡅࡿ⫢⮚ࢢࣝࢱࢳ࢜ࣥ㔞

ྛⅬࡣྛಶయࡢࢢࣝࢱࢳ࢜ࣥ㔞ࢆ♧ࡋ, ᶓᲬࡣྛ⩌ࡢᖹᆒ㸦୰ኸ㸧࠾ࡼࡧᶆ‽೫ᕪ㸦ୖୗ㸧ࢆ, ᣓᘼࡣ᥇ᮦ ᫬㛫ࢆ♧ࡍࠋALF: ⮬⏤ᦤ㣵ୗ, Fasting: 16 ᫬㛫⤯㣗ୗ, RF: ไ㝈⤥㣵ୗ㸦⮬⏤ᦤ㣵ࡢ 70%㸧ࠋ **: p < 0.01 vs ALF ⩌ (9:00)㸦Dunnett ࡢከ㔜ẚ㍑᳨ᐃ㸧ࠋ##_{: p < 0.01 vs RF ⩌ (9:00)㸦Student ࡢ t ᳨ᐃ㸧ࠋ}

36

ᅗ2-6. ␗࡞ࡿᦤ㣵᮲௳ୗ࡟࠾ࡅࡿ APAP ᢞ୚㸦800 mg/kg య㔜㸧24 ᫬㛫ᚋࡢ⫢⤌⧊ീ

㸦A㸧⮬⏤ᦤ㣵ୗ 9:00 ᢞ୚: ⫢⤌⧊࡟᫂ࡽ࠿࡞ኚ໬ࡣࡳࡽࢀ࡞࠸ࠋ㸦B㸧⮬⏤ᦤ㣵ୗ 16:00 ᢞ୚: ⅖⑕ᛶ⣽⬊ ᾐ₶ࢆకࡗࡓᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀࡓࠋ㸦C㸧16 ᫬㛫⤯㣗ୗ 9:00 ᢞ୚: ⅖⑕ᛶ⣽⬊ᾐ₶ࢆకࡗࡓ ᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀࡓࠋ㸦D㸧ไ㝈⤥㣵᮲௳ୗ 16:00 ᢞ୚: ୰ᚰ㟼⬦࿘ᅖ࡟⅖⑕ᛶ⣽⬊ᾐ₶ࢆక ࡗࡓ༢⣽⬊ᛶࡢ⫢⣽⬊ቯṚࡀほᐹࡉࢀࡓࠋ▮༳ࡣ༢⣽⬊ᛶࡢ⫢⣽⬊ቯṚࢆ♧ࡍࠋCV: central vein. PT: Portal triad. H.E.ᰁⰍ (A, B, C: ×100, D: ×400)ࠋ

37

ᅗ2-7. ␗࡞ࡿᦤ㣵᮲௳ୗ࡟࠾ࡅࡿᅄሷ໬Ⅳ⣲ᢞ୚㸦800 mg/kg య㔜㸧24 ᫬㛫ᚋࡢ⫢⤌⧊ീ

㸦A㸧a: ⮬⏤ᦤ㣵ୗ 10:00 ᢞ୚: ⅖⑕ᛶ⣽⬊ᾐ₶ࢆకࡗࡓᑠⴥ୰ᚰᛶࡢ⫢⣽⬊ቯṚ࡜, ୰㛫ᇦ࡛⬡⫫ኚᛶࡀ ほᐹࡉࢀࡓࠋb: 16 ᫬㛫⤯㣗ୗ 10:00 ᢞ୚: ᑠⴥ୰ᚰᇦ࠿ࡽ୰㛫ᇦ࡟࠿ࡅ࡚⅖⑕ᛶ⣽⬊ᾐ₶ࢆకࡗࡓ⫢⣽⬊ቯ Ṛ࡜, ୰㛫ᇦ࡛⬡⫫ኚᛶࡀほᐹࡉࢀࡓࠋCV: central vein. PT: Portal triad. H.E.ᰁⰍ (×100)ࠋ㸦B㸧⮬⏤ᦤ㣵 ୗ࠾ࡼࡧ16 ᫬㛫⤯㣗ୗ࡟࠾ࡅࡿ ALT ್࠾ࡼࡧ AST ್ࠋ⦪Წࡣྛ⩌ࡢᖹᆒ࠾ࡼࡧᶆ‽೫ᕪࢆ♧ࡍࠋALF: ⮬ ⏤ᦤ㣵ୗ, Fasting: 16 ᫬㛫⤯㣗ୗࠋ*p < 0.05 vs ALF ⩌㸦Welch ࡢ t ᳨ᐃ㸧ࠋ

38 ᅗ2-8. ⫢⣽⬊ቯṚ࡟ᑐࡍࡿ EEBGP ᢞ୚ࡢᙳ㡪

㸦A㸧APAP㸦800 mg/kg య㔜㸧ᢞ୚ 24 ᫬㛫ᚋࡢ⫢⤌⧊ീࠋa: Control ⩌, b: EEBGP ⩌ࠋCV: central vein. PT: Portal triad. H.E.ᰁⰍ (×100)ࠋ㸦B㸧Control ⩌࠾ࡼࡧ EEBGP ⩌࡟࠾ࡅࡿ⫢⣽⬊ቯṚࡢ⠊ᅖࠋ⦪Წࡣྛ ⩌ࡢᖹᆒ࠾ࡼࡧᶆ‽೫ᕪࢆ♧ࡍࠋ*p < 0.05 vs control ⩌㸦Student ࡢ t ᳨ᐃ㸧ࠋ㸦C㸧Control ⩌࠾ࡼࡧ EEBGP ⩌࡟࠾ࡅࡿALT ್࠾ࡼࡧ AST ್ࠋ⦪Წࡣྛ⩌ࡢᖹᆒ࠾ࡼࡧᶆ‽೫ᕪࢆ♧ࡍࠋ

39 ᅗ2-9. ⫢⮚࡟࠾ࡅࡿᢠ㓟໬㓝⣲㑇ఏᏊࡢⓎ⌧㔞

⦪Წࡣྛ⩌ࡢᖹᆒ࠾ࡼࡧᶆ‽೫ᕪࢆ♧ࡍࠋྛ㑇ఏᏊࡢⓎ⌧㔞ࡣGapdhࡢⓎ⌧㔞࡛⿵ṇࡋࡓࠋ

ᅗ2-10. ⫢⮚࡟࠾ࡅࡿ⅖⑕㛵㐃㑇ఏᏊࡢⓎ⌧㔞

A: ࢧ࢖ࢺ࢝࢖ࣥ, B: ࢣࣔ࢝࢖ࣥ࠾ࡼࡧ୍㓟໬❅⣲ྜᡂ㓝⣲ࠋ⦪Წࡣྛ⩌ࡢᖹᆒ࠾ࡼࡧᶆ‽೫ᕪࢆ♧ࡍࠋྛ 㑇ఏᏊࡢⓎ⌧㔞ࡣGapdhࡢⓎ⌧㔞࡛⿵ṇࡋࡓࠋ*: p < 0.05 vs Control ⩌㸦Student ࡢ t ᳨ᐃ㸧ࠋ

40

ᅗ2-11. ⫢⣽⬊ቯṚࡢ⠊ᅖ࡜⫢⮚࡟࠾ࡅࡿ⅖⑕㛵㐃㑇ఏᏊࡢⓎ⌧㔞ࡢ㛵ಀ

⦪㍈࡟ྛ㑇ఏᏊࡢⓎ⌧㔞ࢆ, ᶓ㍈࡟⫢⣽⬊ቯṚࡢ⠊ᅖࢆ♧ࡍࠋ㸦A㸧Il1b㑇ఏᏊ, 㸦B㸧Il6㑇ఏᏊ, 㸦C㸧Il10

㑇ఏᏊ, 㸦D㸧Ccl3㑇ఏᏊ, 㸦E㸧Cxcl2㑇ఏᏊࠋྛ㑇ఏᏊࡢⓎ⌧㔞ࡣGapdhࡢⓎ⌧㔞࡛⿵ṇࡋࡓࠋR ್㸦┦ 㛵ಀᩘ㸧࠾ࡼࡧp್㸦┦㛵ࡢ᭷ពᛶ㸧ࡣSpearman ࡢ㡰఩┦㛵ಀᩘࢆ⏝࠸࡚⟬ฟࡋࡓࠋ

41

ᅗ2-12. ᅄሷ໬Ⅳ⣲࡟ࡼࡿ⫢⣽⬊ቯṚ࡟ᑐࡍࡿ EEBGP ᢞ୚ࡢᙳ㡪

㸦A㸧ᅄሷ໬Ⅳ⣲㸦800 mg/kg య㔜㸧ᢞ୚ 24 ᫬㛫ᚋࡢ⫢⤌⧊ീࠋa: Control ⩌, b: EEBGP ⩌ࠋCV: central vein. PT: Portal triad. H.E.ᰁⰍ (×100)ࠋ㸦B㸧Control ⩌࠾ࡼࡧ EEBGP ⩌࡟࠾ࡅࡿ ALT ್࠾ࡼࡧ AST ್ࠋ ⦪Წࡣྛ⩌ࡢᖹᆒ࠾ࡼࡧᶆ‽೫ᕪࢆ♧ࡍࠋ

42

➨

➨

2 ❶

43 ࠙ㅖゝࠚ ࢔࣑ࣟ࢖ࢻ A㸦AA㸧࢔࣑ࣟ࢖ࢻ⑕ࡣ, 㛵⠇࣐ࣜ࢘ࢳࡸࢡ࣮ࣟࣥ⑓࡞࡝ࡢ៏ᛶ⅖⑕⑌ᝈ࡟కࡗ࡚⫢⮚࡟ ࡚⏘⏕ࡉࢀࡿ⅖⑕ᛶࢱࣥࣃࢡ㉁ࡢ1 ࡘ, ⾑Ύ࢔࣑ࣟ࢖ࢻ A㸦SAA㸧ࢱࣥࣃࢡࡀኚᛶࡋ࡚㔜ྜࡋᙧᡂࡉࢀࡿ ࢔࣑ࣟ࢖ࢻA ⧄⥔ࡀ, ⭁⮚, ⫢⮚, ⭈⮚, ⭠⟶➼ࡢ඲㌟⮚ჾࡸ㛵⠇࡟ỿ╔ࡋ࡚⭈୙඲ࡸᾘ໬⟶ᶵ⬟㞀ᐖࢆ Ⓨ⑕ࡋ, 㔜ᗘࡢሙྜ࡟ࡣṚ࡟⮳ࡿ⑌ᝈ࡛࠶ࡿ[97, 98]ࠋAA ࢔࣑ࣟ࢖ࢻ⑕ࡢⓎ⑕࡟ࡣ, ⅖⑕่⃭࡟ࡼࡿᣢ⥆ ⓗ࡞๓㥑ࢱࣥࣃࢡࡢ⏕ᡂ࡜, ࡑࢀ࡟⥆ࡃ࢔࣑ࣟ࢖ࢻ A ࡢኚᛶࡀ㘽࡜࡞ࡿ[98]㸦ᅗ 3-1㸧ࠋᢠ IL-6 ཷᐜయᢠ యࡢᢞ୚࡟ࡼࡾ៏ᛶ⅖⑕ᝈ⪅࡛ቑຍࡋࡓ⾑୰ SAA 㔞ࢆṇᖖ್ࡲ࡛ᡠࡍࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࡀ[99], SAA ࡢኚᛶࡸ㔜ྜࡢཎᅉࡣ᫂ࡽ࠿࡟࡞ࡗ࡚ࡣ࠾ࡽࡎ, AA ࢔࣑ࣟ࢖ࢻ⑕࡟ᑐࡍࡿ἞⒪ἲࡣ☜❧ࡉࢀ࡚࠸࡞࠸ [100, 101]ࠋ࣐࢘ࢫࢆ⏝࠸ࡓᐇ㦂࡛ࡣ, ࢔࣑ࣟ࢖ࢻ β㸦Aβ㸧ࡢỿ╔ࢆࣅࢱ࣑ࣥ E [102]ࡸ α-࣏ࣜ㓟[103], ࢡ ࣝࢡ࣑ࣥ[104, 105]➼ࡢᢠ㓟໬≀㉁ࡀᢚไࡋ, in vitro࡟࠾࠸࡚ࢡࣝࢡ࣑ࣥࡀAβ ⥺⥔ࡢᙧᡂࢆᢚไࡍࡿࡇ ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[106]ࠋࡲࡓ, ᐙ᪘ᛶ࢔࣑ࣟ࢖ࢻ࣏ࣜࢽ࣮ࣗࣟࣃࢳ࣮࡛ࡶࢺࣛࣥࢫࢧ࢖ࣞࢳࣥỿ╔ࡀ ᢠ㓟໬≀㉁࡟ࡼࡾᢚไࡉࢀ[107], ⮅ᓥ࢔࣑ࣟ࢖ࢻỿ╔⑕ࡢ⑓ែ㐍⾜࡟㓟໬ࢫࢺࣞࢫࡀ㛵ࢃࡿࡇ࡜ࡀሗ࿌ ࡉࢀ࡚࠸ࡿ[108]ࠋAA ࢔࣑ࣟ࢖ࢻ⑕ᝈ⪅࡛ࡶ⮚ჾ୰ࡢ㐣㓟໬⬡㉁ࡢቑຍࡀሗ࿌ࡉࢀ࡚࠸ࡿࡀ[109], 㓟໬ ࢫࢺࣞࢫ࡜AA࢔࣑ࣟ࢖ࢻ⑕ࡢ⑓ែ࡜ࡢ㛵ಀࡸ AA ࢔࣑ࣟ࢖ࢻ⑕࡟࠾ࡅࡿᢠ㓟໬≀㉁ࡢᦤྲྀࡀ࢔࣑ࣟ࢖ࢻ ⧄⥔ࡢᙧᡂ࡟୚࠼ࡿᙳ㡪࡟ࡘ࠸࡚ࡣ᫂ࡽ࠿࡟࡞ࡗ࡚࠸࡞࠸ࠋ ඛ⾜◊✲࡛ࡣ, ࣐࢘ࢫ࡟࠾࠸࡚, ࣏ࣉࣛ⣔ࣉ࣏ࣟࣜࢫ࡟ྵࡲࢀࡿࣆࣀࢭࣥࣈࣜࣥࡀ Aβ ⧄⥔ࡢᙧᡂ࡜⚄ ⤒ࡢኚᛶࢆᢚไࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[110]ࠋࡲࡓ, EEBGP[111]ࡸ BGP ࡟ྵࡲࢀࡿ 3, 4, 5-ࢺࣜ- O-࢝ࣇ࢙࢜࢖ࣝ࢟ࢼ㓟[112]ࡀ, in vitro࡟࡚⚄⤒⣽⬊ࢆAβ ࡟ࡼࡿẘᛶ࠿ࡽಖㆤࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋ ᮏ❶࡛ࡣ, ⅖⑕≧ែୗ࡛࢔࣑ࣟ࢖ࢻ A ⥺⥔ࢆᢞ୚ࡍࡿࡇ࡜࡟ࡼࡾ⫢⮚ࡸ⭁⮚࡟࢔࣑ࣟ࢖ࢻ⥺⥔ࡀỿ╔ ࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ[113, 114]࣐࢘ࢫ AA ࢔࣑ࣟ࢖ࢻ⑕ࣔࢹࣝࢆ⏝࠸࡚, EEBGP ࡢΰ㣵ᢞ୚࡟ࡼࡿ ಖㆤຠᯝࢆ᳨ウࡍࡿ࡜࡜ࡶ࡟, ᢠ㓟໬㓝⣲࠾ࡼࡧ⅖⑕㛵㐃㑇ఏᏊ࡜ࡢ㛵ಀ࡟ࡘ࠸࡚☜ㄆࡋࡓࠋ ࠙ᮦᩱ࡜᪉ἲࠚ ࣭⿕㦂≀㉁  ⿕㦂≀㉁࡟ࡣ, EEBGP ࢆ⏝࠸ࡓࠋEEBGP ࢆࢹ࢟ࢫࢺࣜࣥ⢊ᮎ࡜ 2:3 ࡢ๭ྜ࡛ΰྜࡋ࡚஝⇱ࡉࡏ, CRF-1㸦Charles river laboratories, ᮾி, ᪥ᮏ㸧࡟ 0.02%, 0.1%࠾ࡼࡧ 0.5%ΰྜࡉࡏࡓࠋ

44 ࣭ື≀

ᐇ㦂࡟ࡣ, 7 㐌㱋, 㞝ࡢ C57BL/6J ࣐࢘ࢫࢆ⏝࠸, ࢥࣥ࣋ࣥࢩࣙࢼࣝ࡞⎔ቃୗ㸦ᐊ  23s1Υ, ‵ᗘ 55s 10㸣, ᫂ᬯྛ 12 ᫬㛫㸦᫂ᮇ㸸8:00㹼20:00㸧࡟ࢥࣥࢺ࣮ࣟࣝࡋࡓື≀ᐊ㸧࡛⣙ 1 㐌㛫㥆໬ࡉࡏࡓᚋ, ヨ㦂 ࡟౑⏝ࡋࡓࠋ࣎ࢺࣝ࡟ධࢀࡓỈ㐨Ỉ࡜⢊ᮎ㣫ᩱ㸦CRF-1; Charles river laboratories㸧ࢆ⮬⏤ᦤྲྀࡉࡏࡓࠋ ඲࡚ࡢື≀ヨ㦂ࡣ, ᮾி㎰ᕤ኱Ꮫෆࡢື≀೔⌮つᐃ࡟ᚑࡗ࡚ᐇ᪋ࡋࡓ㸦ᢎㄆ␒ྕ㸸#27-96㸧ࠋ ࣭ᐇ㦂ࢹࢨ࢖ࣥ ࣐࢘ࢫࢆ4 ⩌࡟ࢢ࣮ࣝࣉศࡅࡋ, ࡑࢀࡒࢀࣉ࣏ࣟࣜࢫࢆ 0%㸦Control ⩌㸧, 0.02%, 0.1%࠾ࡼࡧ 0.5%ࡢ ⏝㔞࡛17 ᪥㛫, ΰ㣵ᢞ୚ࡋࡓࠋᢞ୚㛤ጞᚋ 7 ᪥┠࡟, ⅖⑕่⃭࡜ࡋ࡚ 2.0%◪㓟㖟Ỉ⁐ᾮ 0.5mL ࢆ⓶ୗ ᢞ୚ࡋ, ྠ᫬࡟ AEF ࢆ 0.3mL ⭡⭍ෆᢞ୚ࡋࡓࠋAEF ࡟ࡣ AA ࢔࣑ࣟ࢖ࢻ⑕⨯ᝈ࣐࢘ࢫࡢ⭁⮚ 20 ಸΰ⃮ ᾮࢆ⏝࠸ࡓࠋELISA ࡟ࡼࡿ SAA ್ ᐃࡢࡓࡵ, ⅖⑕่⃭ 3 ᪥ᚋ࡟║ᗏ᥇⾑ࡋࡓࠋ⅖⑕่⃭ᚋ 10 ᪥࡟ࢪ ࢚ࢳ࢚࣮ࣝࢸࣝ㯞㓉ୗ࡛ᚰ⮚࠿ࡽ඲᥇⾑ࡋࡓᚋ, ๗᳨ࡋࡓࠋᮏࣔࢹࣝ࡟࠾࠸࡚, ࢔࣑ࣟ࢖ࢻỿ╔ࡀሗ࿌ࡉ ࢀ࡚࠸ࡿ⭁⮚࠾ࡼࡧ⫢⮚ࢆ᥇ᮦࡋ, ࣓ࢱ࣮࢝ࣥᾮ࡛ᅛᐃࡋ, H.E.ᰁⰍ࠾ࡼࡧච␿⤌⧊໬Ꮫࢆ⾜ࡗࡓࠋ⫢⮚ ࡢ୍㒊ࡣᾮయ❅⣲࡛෾⤖ࡋ, ᢠ㓟໬㓝⣲࠾ࡼࡧ⅖⑕㛵㐃ᅉᏊࡢ㑇ఏᏊⓎ⌧㔞ࢆ ᐃࡋࡓࠋ ࣭ච␿⤌⧊໬Ꮫ

⭁⮚࠾ࡼࡧ⫢⮚࡟࠾࠸࡚ᢠ࣐࢘ࢫSAA ࣖࢠᢠయ㸦1㸸160; R&D Systems, Minneapolis, MN, USA㸧 ࢆ⏝࠸࡚ච␿⤌⧊໬Ꮫࢆ⾜ࡗࡓࠋ࢔࣑ࣟ࢖ࢻỿ╔㔞ࡣ, Image J ࢆ⏝࠸࡚, 㢧ᚤ㙾㸦™50 ಸ㸧ୗ࡛ࣛࣥࢲ

࣒࡟3 ᯛྲྀᚓࡋࡓ⏬ീ࠿ࡽ⤌⧊㠃✚࡟࠾ࡅࡿᰁⰍ㝧ᛶ㒊఩ࡢ๭ྜࢆ ᐃࡋࡓࠋ

࣭⾑୰SAA ࡢ ᐃ

 ⅖⑕཯ᛂ 3 ᪥ᚋ࠾ࡼࡧ 10 ᪥ᚋࡢ⾑ᾮ࡟ࡼࡾᚓࡽࢀࡓ⾑Ύ࠿ࡽ, ࣐࢘ࢫ SAA  ᐃ ELISA ࢟ࢵࢺ 㸦Tridelta Development Limited, Maynooth, County Kildare, Ireland㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋ

࣭ᐃ㔞PCR

 Control ⩌࠾ࡼࡧ EEBGP ⩌ࡼࡾᚓࡽࢀࡓ⫢⮚⣙ 50 mg ࡟ RNeasy Mini Kit (Qiagen, Venlo Netherlands)ࢆຍ࠼࡚࣍ࣔࢪࢼ࢖ࢬ㸦12, 000 rpm㸧ࡋࡓࠋᢳฟࡋࡓ Total RNA ࠿ࡽ, PrimeScript RT