植物乳酸菌による生薬発酵で生ずる抗炎症性物質の構造と機能に関する研究

概要

༤ኈㄽᩥ

᳜≀ங㓟⳦࡟ࡼࡿ⏕⸆Ⓨ㓝࡛⏕ࡎࡿᢠ⅖⑕ᛶ≀㉁ࡢ
ᵓ㐀࡜ᶵ⬟࡟㛵ࡍࡿ◊✲

2020 ᖺ
ᗈᓥ኱Ꮫ኱Ꮫ㝔་ṑ⸆ಖ೺Ꮫ◊✲⛉
ᖹᡂ 28 ᖺᗘධᏛ
ᒸ

ᮏ

▱

Ꮚ

࠙┠ḟࠚ
ᗎ❶࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭1
➨୍❶ ᳜≀ங㓟⳦ࢆ⏝࠸ࡓⓎ㓝Ⱅⴥᢳฟ≀ࡢᢠ⅖⑕స⏝
➨୍⠇ ⫢⒴⏤᮶⣽⬊ᰴࢆ⏝࠸ࡓ NASH ࣔࢹࣝ⣽⬊ࡢసᡂ ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭ 4
➨஧⠇ NASH ࣔࢹࣝ⣽⬊ࢆ⏝࠸࡚ࡢⓎ㓝⏕⸆ᢳฟ≀ࡢᢠ⅖⑕స⏝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭6
➨୕⠇ ␗࡞ࡿ᳜≀ங㓟⳦࡛Ⓨ㓝ࡉࡏࡓⰕⴥᢳฟ≀ࡢᢠ⅖⑕స⏝࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭8
➨ᅄ⠇ ᳜≀ங㓟⳦ࡢಖ᭷ࡍࡿE-ࢢࣝࢥࢩࢲ࣮ࢮ㑇ఏᏊࡢゎᯒ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭9
➨஬⠇ ࡲ࡜ࡵ࡜⪃ᐹ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭12
➨஧❶ ᢠ⅖⑕άᛶ≀㉁ࡢ⢭〇࡜໬Ꮫᵓ㐀ࡢỴᐃ
➨୍⠇ Ⓨ㓝࠾ࡼࡧᮍⓎ㓝Ⱅⴥᢳฟ≀࠿ࡽάᛶ≀㉁ࡢᢳฟ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭13
➨஧⠇ HPLC ࢆ⏝࠸ࡓⓎ㓝࠾ࡼࡧᮍⓎ㓝Ⱅⴥᢳฟ≀ࡢᡂศศᯒ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭16
➨୕⠇ Ⓨ㓝Ⱅⴥᢳฟ≀࠿ࡽάᛶ≀㉁ࡢศ㞳⢭〇࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭18
➨ᅄ⠇ άᛶ≀㉁ࡢᵓ㐀Ỵᐃ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭22
➨஬⠇ ྛ⏬ศࡢ HPLC ศᯒ࡜άᛶホ౯࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭26
➨භ⠇ ࡲ࡜ࡵ࡜⪃ᐹ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭29
⥲ᣓ࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭32
ᐇ㦂ࡢ㒊࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭34
1.

౑⏝⳦ᰴ

2.

౑⏝⣽⬊

3.

౑⏝⏕⸆

4.

౑⏝ヨ⸆

5.

౑⏝ᇵᆅ

6.

➨୍❶ࡢᐇ㦂

7.

➨஧❶ࡢᐇ㦂

8.

⤫ィゎᯒ

ཧ⪃ᩥ⊩࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭39
ㅰ㎡࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭࣭44

[␎
␎ㄒ୍ぴ]
1

H NMR

13

C NMR

proton nuclear magnetic resonance
carbon-13 nuclear magnetic resonance

B.

Bifidobacterium

CC

column chromatography

D-MEM

dulbecco's modified eagle's medium

DMSO

dimethyl sulfoxide

EDTA

ethylenediaminetetraacetic acid

ELISA

enzyme-linked immunosorbent assay

FA

fatty acid

FBS

fetal bovine serum

GC-EI-MS

gas chromatography electron ionization mass spectrometry

HPLC

high performance liquid chromatography

HR-EI-MS

high-resolution electrospray ionization mass spectrometry

IL

interleukin

LAB

lactic acid bacteria

Lb.

Lactobacillus

MeOH

methanol

MRS

de Man, Rogosa, and Sharpe

MS

mass spectrum

NASH

non-alcoholic steatohepatitis

NF-NB

nuclear factor-kappa B

ODS

octadecylsilanized silica gel

ORF

open reading frame

P.

Pediococcus

PBS

phosphate buffered saline

SQL

sesquiterpene lactone

TG

triglyceride

TLR

toll like receptor

[ᗎ
ᗎ❶]
᪥ᮏ⸆ᒁ᪉࡛ࡣ, ⏕⸆ࢆࠕື᳜≀ࡢ⸆⏝࡜ࡍࡿ㒊ศ, ⣽⬊ෆᐜ≀, ศἪ≀, ᢳฟ≀ࡲࡓࡣ㖔
≀࡞࡝ࠖ࡜ᐃ⩏ࡉࢀ࡚࠸ࡿࠋࡍ࡞ࢃࡕ, ⏕⸆ࡣ, ⸆ຠᡂศࢆ༢୍≀㉁࡜ࡏࡎ࡟⏝࠸ࡿࠕᮍ⢭〇
ࡢࢡࢫ࡛ࣜࠖ࠶ࡿࠋ1–3 ୡ⣖࡟୰ᅜ࡛⦅⧩ࡉࢀࡓ᭱ྂࡢ⸆≀Ꮫࡢ᭩ࠕ⚄㎰ᮏⲡ⤒ࠖ࡟ࡣ, 365 ✀
ࡢ⏕⸆ࡀ཰㍕ࡉࢀ, ࡑࡢෆヂࡣ, ᳜≀⸆ࡀ 252 ✀, ື≀⸆ࡀ 67 ✀, 㖔≀⸆ࡀ 46 ✀࡛࠶ࡿࠋᮏ
᭩࡟ࡼࢀࡤ, ⚄㎰ࡣ⮬ࡽẘ࿡࡟ࡼࡾ⏕⸆ࡢຠ⬟ࡸᏳ඲ᛶࢆ☜ㄆࡋ࡚ࡁࡓ࡜ࡉࢀࡿࠋ⏕⸆ࢆ⤌
ࡳྜࢃࡏ࡚సࡽࢀࡿࡢࡀ, ₎᪉⸆࡛࠶ࡿࠋ₎᪉⸆ࢆᢅ࠺₎᪉་Ꮫࡣ, 5–6 ୡ⣖ࡈࢁ࡟୰ᅜ་Ꮫ
ࢆᇶ࡟᪥ᮏ࡛Ⓨᒎࡋࡓఏ⤫་Ꮫࢆᣦࡍࠋ≉࡟ྂ඾་Ꮫ᭩࡟ᇶ࡙ࡃ⸆≀⒪ἲࢆ₎᪉་Ꮫ࡜࿧ࡧ,
₎᪉⸆ࡣ, ࠕയᐮ㞧⑓ㄽࠖ(⌧ᅾ࡛ࡣ, ࠕയᐮㄽࠖཬࡧࠕ㔠༄せ␎ࠖࡢࡇ࡜ࢆᣦࡍ) ࡟グ㍕ࡉࢀ
࡚࠸ࡿࠋࡇࡢࡼ࠺࡟, ⏕⸆ (₎᪉⸆) ࡣ, ࣄࢺ࡬ࡢᢞ୚࡟ࡼࡾࡑࡢ᭷ຠᛶࡸᏳ඲ᛶࡀホ౯ࡉࢀ
࡚ࡁࡓ, Ṕྐⓗ࡟ࡶ㧗࠸ရ㉁ࡢ⸆๣࡛࠶ࡿࠋ
⏕≀άᛶࢆ♧ࡍࡶࡢࢆྵࡵ, ✀ࠎࡢ໬ྜ≀ࢆྵࡴኳ↛≀࡟࠾࠸࡚, άᛶ≀㉁ࡢᮏయࢆ᫂ࡽ
࠿࡟ࡍ࡭ࡃ, ศ㞳᥈⣴◊✲ࡀ㐍ࡵࡽࢀ࡚ࡁࡓࠋ౛࠼ࡤ, ࢣࢩ࠿ࡽࡣࣔࣝࣄࢿࡀ༢㞳ࡉࢀ, ࢱࣂ
ࢥ࠿ࡽࡣࢽࢥࢳࣥ, ࢥ࣮ࣄ࣮࠿ࡽ࢝ࣇ࢙࢖ࣥ, ࢣࢪࢠࢱࣜࢫ࠿ࡽࡣࢪࢦ࢟ࢩࣥ࡜, ⏕⸆ࢆࡣ
ࡌࡵ࡜ࡍࡿኳ↛≀࠿ࡽࡣᩘከࡃࡢ⏕≀άᛶ≀㉁ࡀ༢㞳ࡉࢀ࡚ࡁࡓࠋࡋࡓࡀࡗ࡚, ⏕⸆ࡣ᪂ࡋ
࠸་⸆ရ࣮ࣜࢻ໬ྜ≀ࡢ㔜せ࡞౪⤥※࡜ࡳ࡞ࡉࢀ࡚࠸ࡿ [1, 2]ࠋࡋ࠿ࡋ࡞ࡀࡽ, ࡑࢀࡽࡢ໬ྜ
≀ࡢ୰࡟ࡣ, ඖࡢ᳜≀࠿ࡽᚓࡽࢀࡿ⏕≀άᛶ≀㉁ࡢ㔞ࡀᑡ࡞࠸ࡇ࡜ࡀࡋࡤࡋࡤㄆࡵࡽࢀࡿ
[3]ࠋ⏕⸆ࡢከࡃࡣ᳜≀࡟⏤᮶ࡋ, ࡑࡢ⏕≀άᛶ≀㉁ࡢከࡃࡣ᳜≀஧ḟ௦ㅰ≀࡛࠶ࡿࠋ᳜≀஧
ḟ௦ㅰ≀ࡣ, ࡑࡢ࢔ࢢࣜࢥࣥࡢỈ⁐ᛶ࡜໬ᏛⓗᏳᐃᛶࢆᨵၿࡍࡿࡓࡵ࡟, ࡑࡢ᳜≀୰࡟㓄⢾
య࡜ࡋ࡚ಖᏑࡉࢀ࡚࠸ࡿ [4]ࠋ㓄⢾యࡣ, ␯Ỉᛶࡀప࠸ࡓࡵ, ࢔ࢢࣜࢥࣥࡼࡾࡶ⏕≀Ꮫⓗ฼⏝
⋡ (bioavailability) ࡀపࡃ, ⤖ᯝ࡜ࡋ࡚ᙅ࠸⏕≀άᛶࡋ࠿♧ࡉ࡞࠸ࡇ࡜ࡀከ࠸ࠋࡍ࡞ࢃࡕ, ᳜
≀஧ḟ௦ㅰ≀ࡀ, ࡑࡢ⏕≀άᛶࢆ᭱኱㝈࡟♧ࡍࡓࡵ࡟ࡣ, άᛶᆺ (࢔ࢢࣜࢥࣥ) ࡬ࡢኚ᥮ࡀ㔜
せ࡛࠶ࡿ࡜ゝ࠼ࡿ [5]ࠋ
⏕⸆ (₎᪉⸆) ࡢ◊✲ࢆ㏻ࡌ࡚, in vivo ࡛⸆ຠࢆ♧ࡍ⏕⸆ࡀ, ᚲࡎࡋࡶ in vitro ࡛ࡣⰾࡋ࠸⤖
ᯝࡀᚓࡽࢀ࡞࠸ࡇ࡜ࡀ⤒㦂ⓗ࡟▱ࡽࢀ࡚࠸ࡓࠋ౛࠼ࡤ, ኱㯤ࡢᡂศ࡛࠶ࡿ sennoside ࡣ,
anthrone ஧㔞య࠿ࡽ࡞ࡿ㓄⢾య࡛࠶ࡿࠋࡑࡢࡓࡵ, ẚ㍑ⓗศᏊࡀ኱ࡁࡃᾘ໬⟶ୖ㒊࡛ࡣ྾཰ࡉ
ࢀࡎ, ᾘ໬⟶ୗ㒊࡟㍺㏦ࡉࢀࡿࠋࡑࡢ㒊఩࡟⏕ᜥࡍࡿ᎘Ẽᛶ⣽⳦࡟ࡼࡗ࡚㓄⢾యࡀຍỈศゎ
ࡉࢀ, ࢔ࢢࣜࢥ࡛ࣥ࠶ࡿ sennidin ࡜࡞ࡗࡓᚋ, ஧㔞యࢆ⧚࠸࡛࠸ࡿⅣ⣲–Ⅳ⣲⤖ྜࡀ㑏ඖⓗ࡟
㛤⿣ࡋ, ༢㔞యࡢ rhein anthrone ࡟ኚ᥮ࡉࢀࡿ [6]ࠋࡇࢀࡽ୍㐃ࡢ sennoside ࡢ rhein anthrone ࡬
ࡢኚ᥮࡟ࡣ Bifidobacterium (B.) sp. SEN, B. dentium, B. adolescentis ࡞࡝࡟ࡼࡿ㓄⢾యࡢẁ㝵ⓗ
ຍỈศゎ, Peptostreptococcus intermedius ࡞࡝ࡢ⣽⳦࡟ࡼࡿ஧㔞యࡢ㛤⿣࡞࡝, ᾘ໬⟶࡟⏕ᜥࡍ
ࡿ⭠ෆ⣽⳦ࡢඹാస⏝ࡀᚲせ࡛࠶ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ [7]ࠋࡲࡓ, Bifidobacterium sp.࠿ࡽ
ࡣ, sennoside ࢆຍỈศゎࡍࡿE-ࢢࣝࢥࢩࢲ࣮ࢮࡀ⢭〇ࡉࢀ࡚࠸ࡿ [8, 9]ࠋศᏊࡢ኱ࡁ࠸
sennoside ࡣ, ௚ࡢ⭠ෆ⣽⳦ࡢᣢࡘE-ࢢࣝࢥࢩࢲ࣮ࢮ࡛ࡣຍỈศゎࡉࢀࡎ, ࡇࡢ Bifidobacterium
sp.ࡀ≉␗ⓗ࡟⏘⏕ࡍࡿE-ࢢࣝࢥࢩࢲ࣮ࢮ࡟ࡼࡾ࢔ࢢࣜࢥࣥ࡜࡞ࡿࡇ࡜ࡀ࡛ࡁࡿࠋࡇࡢሗ࿌ࡣ,



㓄⢾యࡢຍỈศゎ཯ᛂ࡟࠾࠸࡚, E-ࢢࣝࢥࢩࢲ࣮ࢮࡢᇶ㉁ㄆ㆑ࡀ㔜せ࡛࠶ࡿࡇ࡜ࢆᙉࡃ♧ࡍ
ࡶࡢ࡛࠶ࡿࠋ
௚᪉, ኱㇋ࡢᡂศ࡛ࣇࣛ࣎ࣀ࢖ࢻࡢ୍✀࡛࠶ࡿ daidzin ࡣ, ⭠ෆ⣽⳦࡟ࡼࡾ daidzein ࡟ኚ᥮
ࡉࢀࡿࠋࡉࡽ࡟ daidzein ࡣ, equol ࡜࠸࠺⏕≀άᛶࡢ㧗࠸໬ྜ≀࡟ኚ᥮ࡉࢀࡿࡇ࡜ࡀሗ࿌ࡉࢀ
࡚࠸ࡿ [10, 11]ࠋࡇࡢࡼ࠺࡟⏕⸆࡞࡝ࡢ᳜≀୰࡟ྵࡲࢀࡿᡂศࡣ, ┿ࡢάᛶ≀㉁࡛ࡣ࡞ࡃ, ࣉ
ࣟࢻࣛࢵࢢࡢᙧ࡛Ꮡᅾࡍࡿࡇ࡜ࡶከ࠸ࠋࡘࡲࡾ, ⭠ෆ⣽⳦࡟ࡼࡾኚ᥮ࡉࢀࡓ᭱⤊ᙧែࡀ⏕⸆
ࡢ⸆⌮άᛶࡢᮏయ࡛࠶ࡿྍ⬟ᛶࡶ㧗ࡃ, ኳ↛⏤᮶ࡢ⸆≀࡜⭠ෆ⣽⳦࡜ࡢᐦ᥋࡞㛵ಀࡀ᫂ࡽ࠿
࡟࡞ࡗ࡚ࡁࡓ [12]ࠋ
⭠ෆ⣽⳦ࡣ, ᳜≀஧ḟ௦ㅰ≀ࢆ⏕≀Ꮫⓗ࡟άᛶ࡞࢔ࢢࣜࢥࣥ࡟ኚ᥮ࡍࡿព࿡࡟࠾࠸࡚, ኱
ࡁ࡞㈉⊩ࢆᯝࡓࡋ࡚࠸ࡿࠋ⭠ෆ࡟࠾ࡅࡿᚤ⏕≀ࡢኚ᥮཯ᛂ࡟ࡣ, ⬺ࢢࣜࢥࢩࣝ໬, ⬺ࣄࢻࣟ࢟
ࢩࣝ໬, 㑏ඖ, C ⎔ษ᩿, ࠾ࡼࡧ⬺࣓ࢳࣝ໬ࡀྵࡲࢀࡿ [10]ࠋ≉࡟, 㓄⢾యࡢຍỈศゎࡣ, ⸆⏝
᳜≀୰ࡢ≀㉁ࡀ⭠ෆኚ᥮ࢆཷࡅࡿ㝿࡟࠾ࡅࡿ㔜せ࡞ࢫࢸࢵࣉ࡛࠶ࡿࠋࢢࣜࢥࢩࢻຍỈศゎ㓝
⣲ࣇ࢓࣑࣮ࣜ࡟ศ㢮ࡉࢀࡿE-ࢢࣝࢥࢩࢲ࣮ࢮ (EC 3.2.1.21) ࡣ, E-ࢢࣜࢥࢩࣝ๓㥑య࠿ࡽ࢔ࢢ
ࣜࢥࣥࢆษࡾ㞳ࡍࡢ࡟㔜せ࡛࠶ࡿࠋࡇࡢ㓝⣲ࡣ, ື≀, ᳜≀, ┿⳦, 㓝ẕ, ⣽⳦࡟ᗈࡃศᕸࡋ
࡚࠸ࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿ [4]ࠋࡋࡓࡀࡗ࡚, E-ࢢࣝࢥࢩࢲ࣮ࢮࢆಖ᭷ࡍࡿᚤ⏕≀࡟ࡼࡿ⏕⸆
ࡢⓎ㓝ࡣ, ⏕≀άᛶ≀㉁ࢆࡑࡢ๓㥑య࠿ࡽኚ᥮ࡍࡿࡓࡵࡢ᭷⏝࡞᪉ἲ࡛࠶ࡿ [13–18]ࠋࡘࡲࡾ,
Ⓨ㓝ᢏ⾡ࡣ, ᳜≀஧ḟ௦ㅰ≀ࡢάᛶࢆ, 㔞ⓗࡶࡋࡃࡣ㉁ⓗ࡟ᨵၿࡍࡿࡓࡵࡢ㨩ຊⓗ࡞࢔ࣉࣟ
࣮ࢳ࡛࠶ࡿ࡜ゝ࠼ࡿࠋ
ங㓟⳦ࡣ, ⭠ෆ⣽⳦ྀࢆᵓᡂࡍࡿ୺せ࡞ᚤ⏕≀ࡢ୍ࡘ࡛࠶ࡿࠋᮡᒣ◊✲ᐊ࡛ࡣ, ᯝ≀, 㔝⳯,
ⰼ, ⸆⏝᳜≀࠿ࡽங㓟⳦ࢆ᥈⣴ศ㞳ࡋ, ࡇࢀࡲ࡛࡟, 1,000 ᰴࢆ㉸࠼ࡿங㓟⳦ (᳜≀ங㓟⳦) ࠿
ࡽᡂࡿࣛ࢖ࣈ࣮ࣛࣜࢆಖ᭷ࡋ࡚࠸ࡿࠋࡑࡋ࡚, ࡇࢀࡽ᳜≀ங㓟⳦ࡢಖ೺ᶵ⬟ᛶࢆホ౯ࡋ, ᗄࡘ
࠿ࡢங㓟⳦ᰴ࡟ࡘ࠸࡚ࡣ඲ࢤࣀ࣒㓄ิࢆゎㄞࡋ࡚࠸ࡿ [19–21]ࠋ
᳜≀ங㓟⳦ࡣ, ࣄࢺࡸື≀࡞࡝ࡢᾘ໬⟶࠿ࡽศ㞳ࡉࢀࡓື≀⏤᮶ࡢங㓟⳦ (ື≀ங㓟⳦)
࡜ࡣ␗࡞ࡿ≉ᛶࢆ᭷ࡋ࡚࠸ࡿࠋ୍ࡘࡣ, ᳜≀ங㓟⳦ࡣ, ⫶㓟ࡸ⫹Ồ࡬ࡢ⪏ᛶࡀື≀ங㓟⳦ࡼࡾ
㧗ࡃ, ⤒ཱྀᦤྲྀࡋ࡚ࡶ⏕⳦≧ែࡢࡲࡲ⭠⟶࡟ᒆࡁࡸࡍ࠸ࠋࡉࡽ࡟ࡣ, ື≀ங㓟⳦ࡣ, ࢔࣑ࣀ㓟
࡞࡝࡟ᑐࡍࡿᰤ㣴せồᛶࡀ㧗ࡃ, ࡑࢀࡽᰤ㣴⣲ࡀ୙㊊ࡋࡓ⎔ቃୗ࡛ࡢ⏕⫱ࡀᅔ㞴࡛࠶ࡿࡢ࡟
ᑐࡋ, ᳜≀ங㓟⳦ࡣ, ࡑࡢࡼ࠺࡞⎔ቃ࡟࠾࠸࡚ࡶⰋዲ࡟⏕⫱࡛ࡁࡿࠋ
ᮏ◊✲࡟ඛ❧ࡗ࡚⾜ࡗࡓணഛᐇ㦂ࡢ⤖ᯝ࠿ࡽ, Ỉ࡛ᢳฟࡋࡓᗄࡘ࠿ࡢ⏕⸆ᢳฟᾮ୰࡛, ᳜
≀ங㓟⳦ Lactobacillus (Lb.) plantarum SN13T ࡣάⓎ࡟ቑṪ࡛ࡁࡿ୍᪉࡛, ື≀ங㓟⳦ࡣࡑࡢ᮲
௳ୗ࡛ࡣ࡯࡜ࢇ࡝ቑṪࡋ࡞࠸ࡇ࡜ࡀ☜ㄆࡉࢀࡓࠋࡇࡢ㐪࠸࡟㛵ࡋ࡚ࡢ⌮⏤ࡢ୍ࡘࡣ, ᳜≀ங
㓟⳦ࡣ, ⮬ᕫࡢ⏘⏕ࡍࡿE-ࢢࣝࢥࢩࢲ࣮ࢮ࡟ࡼࡾ, ⏕⫱࡟ᚲせ࡞⢾※࡜ࡋ࡚, ᳜≀ᡂศ࡛࠶ࡿ
ࢭࣟࣅ࣮࢜ࢫ (ࢢࣝࢥ࣮ࢫ 2 ศᏊࡀE-1, 4 ⤖ྜࡋࡓࡶࡢ) ࡸ᳜≀஧ḟ௦ㅰ≀ (E-ࢢࣝࢥࢩࢻ)
࡞࡝ࢆຍỈศゎࡋ࡚ᚓࡓ⢾ࢆ฼⏝࡛ࡁࡿࡓࡵ࡜⪃࠼࡚࠸ࡿࠋࡲࡓࡶ࠺୍ࡘࡢ⌮⏤࡜ࡋ࡚, ᳜
≀ங㓟⳦ࡣ, ⏕⸆ᢳฟᾮ୰࡟ࡋࡤࡋࡤ᳨ฟࡉࢀࡿᢠ⳦≀㉁࡟ᑐࡍࡿ⪏ᛶࢆ᭷ࡍࡿࡇ࡜ࡀ⪃࠼
ࡽࢀࡿࠋࡇࡢࡼ࠺࡟, ᳜≀ங㓟⳦ࢆ⏕⸆Ỉᢳฟᾮ୰࡛ᇵ㣴ࡋࡓሙྜ࡟ࡣ, ங㓟⳦ࡀቑṪࡍࡿ㐣
⛬࡛⏕ᡂࡋࡓ࢔ࢢࣜࢥࣥࢆᚓࡿࡇ࡜ࡀ࡛ࡁࡿྍ⬟ᛶࡀ࠶ࡿࠋ
ᮏ◊✲࡛ࡣ, ࠕ᳜≀ங㓟⳦ࢆ⏝࠸ࡓⓎ㓝࡟ࡼࡗ࡚, ⏕⸆୰ࡢ≀㉁࠿ࡽ᪂ࡓ࡞⏕≀άᛶ≀㉁ࢆ



⏕ᡂࡉࡏࡿࠖ࡜࠸࠺௬ㄝࢆ❧࡚, ࡑࢀࢆ᳨ドࡍࡿࡇ࡜ࢆ┠ⓗ࡜ࡋࡓࠋࡲࡓ, ⏕⸆ࡢⓎ㓝ࣉࣟࢭ
ࢫ࡟࠾࠸࡚㔜せ࡞཯ᛂ࡛࠶ࡿ㓄⢾యຍỈศゎ㓝⣲E-ࢢࣝࢥࢩࢲ࣮ࢮ࡟╔┠ࡋ, ඲ࢤࣀ࣒ゎᯒ
ࢆ⾜ࡗࡓ᝟ሗ࡟ᇶ࡙ࡁ, SN13T ᰴࡢಖ᭷ࡍࡿE-ࢢࣝࢥࢩࢲ࣮ࢮ㑇ఏᏊࢆ, ௚ࡢ᳜≀ங㓟⳦ࡢࡑ
ࢀ࡜ẚ㍑ࡋࡓࠋ



[➨
➨୍❶]
᳜≀ங㓟⳦ࢆ⏝࠸ࡓⓎ㓝Ⱅⴥᢳฟ≀ࡢᢠ⅖⑕స⏝
Ⓨ㓝⏕⸆ᢳฟ≀ࡢ⏕≀άᛶࡣ, Chavez-Tapia ➼ࡀሗ࿌ࡋࡓ㠀࢔ࣝࢥ࣮ࣝᛶ⬡⫫ᛶ⫢⅖ (nonalcoholic steatohepatitis: NASH) ࣔࢹࣝ⣽⬊ࢆ⏝࠸࡚, ᢠ⅖⑕άᛶࢆホ౯ࡍࡿࡇ࡜࡛⾜ࡗࡓ
[22]ࠋࡇࡢࣔࢹ࡛ࣝࡣ, ⬡⫫㓟 (fatty acids: FAs) ࢆᢞ୚ࡍࡿ࡜, HuH-7 ⣽⬊࠿ࡽ㐟㞳ࡉࢀࡿ࢖
ࣥࢱ࣮ࣟ࢖࢟ࣥ (interleukin: IL)-8 㔞ࡀ⣙ 2 ಸቑຍࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋࡼࡗ࡚Ⓨ㓝⏕
⸆ᢳฟ≀ࡢᢠ⅖⑕άᛶࢆ FA ฎ⌮ࡋࡓ HuH-7 ⣽⬊ᇵ㣴ᾮ୰࡟ᨺฟࡉࢀࡓ IL-8 ࡢ㔞࡟ᑐࡍࡿ㜼
ᐖᗘ࡛ホ౯ࡍࡿࡇ࡜࡜ࡋࡓࠋ
ෆ⮚⫧‶࡟㛵㐃ࡍࡿ⑌ᝈࡣ࣓ࢱ࣎ࣜࢵࢡࢩࣥࢻ࣮࣒ࣟ࡟ศ㢮ࡉࢀ, NASH ࡶࡑࡢ୍ࡘ࡛࠶
ࡿࠋNASH ࡛ࡣ, ᰤ㣴㐣๫࡟ࡼࡾ⵳✚ࡋࡓ㐟㞳 FA ࡀ⫢⮚ࡢ TLR4/2 ࢆάᛶ໬ࡋ, ⥆ࡃ NF-NB
࢝ࢫࢣ࣮ࢻࡢάᛶ໬ࢆ௓ࡋ, ⫢⣽⬊࠿ࡽࡢ IL-8 ࡢศἪࢆಁ㐍ࡍࡿ [23]ࠋIL-8 ࡣ, ᙉຊ࡞ዲ୰
⌫㉮໬ᅉᏊ࡛࠶ࡾ, ࡲࡓ NASH ᝈ⪅࡛ቑຍࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ [24, 25]ࠋዲ୰⌫ᾐ₶ࡣ
NASH ࡢ≉ᚩࡢ୍ࡘ࡛࠶ࡾ [26], ᾐ₶ࡋࡓዲ୰⌫࡟ࡼࡾ⅖⑕ࡀࡉࡽ࡟៮ᝏࡍࡿࡓࡵ, NASH ࡢ
Ⓨ⑕࡜㐍⾜࡟࠿࠿ࢃࡿ㔜せ࡞ᅉᏊࡢ୍ࡘ࡛࠶ࡿࠋຍ࠼࡚ IL-8 ࢩࢢࢼࣝఏ㐩⤒㊰ࡢ㜼ᐖࡣ, ື
≀ࣔࢹ࡛ࣝ⫢⮚ࡢᦆയࢆῶࡽࡍࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿ [27, 28]ࠋ
ᮏ❶࡛ࡣ, ࡲࡎ NASH ࣔࢹࣝ⣽⬊ࢆᩥ⊩ [22] ࡟ᚑ࠸స〇ࡋࡓࠋࡑࡋ࡚ᮡᒣ◊✲ᐊࡀಖᏑࡍ
ࡿ᳜≀ங㓟⳦ࡢ୍ࡘ࡛࠶ࡾ, ᗄࡘ࠿ࡢ⏕⸆Ỉᢳฟᾮ୰࡛ቑṪᛶࡢ㧗࠸ Lb. plantarum SN13T ࡛
⏕⸆ᢳฟ≀ࢆⓎ㓝ࡉࡏࡓࠋࡑࢀࡽࡢⓎ㓝⏕⸆ᢳฟ≀࡟Ꮡᅾࡍࡿᢠ⅖⑕άᛶࢆୖグ NASH ࣔࢹ
ࣝ⣽⬊ࡢ⣔ࢆ⏝࠸࡚ ᐃࡋࡓࠋࡑࡢ⤖ᯝ, ⏕⸆ࡢ୍ࡘࠕⰕⴥ (࢞࢖ࣚ࢘) ࠖࡢⓎ㓝ᢳฟ≀ࡀ, ᢠ
⅖⑕άᛶࢆ♧ࡍࡇ࡜ࡀ᫂ࡽ࠿࡟࡞ࡗࡓࠋࡲࡓ, SN13T ᰴ௨እࡢ඲ࢤࣀ࣒ࢆゎㄞࡋࡓ 2 ᰴࢆ⏝
࠸, ࡑࢀࡒࢀస〇ࡋࡓⓎ㓝Ⱅⴥᢳฟ≀ࡢຠᯝࢆ SN13T ᰴࡢࡑࢀ࡜ẚ㍑ࡋࡓࠋ

➨୍⠇ ⫢⒴⏤᮶⣽⬊ᰴࢆ⏝࠸ࡓ NASH ࣔࢹࣝ⣽⬊ࡢసᡂ
NASH ࣔࢹࣝ⣽⬊ࡣ, Chavez-Tapia ➼ࡢሗ࿌࡟ᇶ࡙࠸࡚స〇ࡋࡓ [22]ࠋ
⫢⒴⏤᮶⣽⬊ᰴ HuH7 ࢆ 24 ࢙࢘ࣝࣉ࣮ࣞࢺ࡟᧛✀ࡋ, 24 ᫬㛫ᚋ࡟ࣃ࣑ࣝࢳࣥ㓟࠾ࡼࡧ࢜ࣞ࢖ࣥ㓟 (ࣔࣝẚ 1 : 2)
ࢆ 600 PM ࡢ⃰ᗘ࡛ῧຍࡋࡓࠋῧຍᚋࡉࡽ࡟ 24 ᫬㛫ᇵ㣴ࡋ, ࢺࣜࢢࣜࢭࣜࢻ (TG) ࢔ࢵࢭ࢖
࢟ࢵࢺࢆ⏝࠸࡚⣽⬊ෆ⬡⫫㔞ࢆ ᐃࡋࡓࠋࡲࡓ, ⣽⬊ᇵ㣴ᾮ୰࡟ᨺฟࡉࢀࡓ IL-8 㔞ࢆ ELISA
ἲ࡟ࡼࡾ᳨ฟࡋࡓࠋࡑࡢ⤖ᯝ, FA ฎ⨨⩌࡛ࡣࢥࣥࢺ࣮ࣟࣝ⩌࡜ẚ㍑ࡋ, ᭷ព࡟⣽⬊ෆ⬡⫫㔞
(Fig. 1) ࠾ࡼࡧ⣽⬊ᇵ㣴ᾮ୰ࡢ IL-8 㔞 (Fig. 2) ࡀ᭷ព࡟ቑຍࡋࡓࠋࡲࡓ FA ฎ⌮࡟ࡼࡾẘᛶࡀ
ฟ࡞࠸ࡇ࡜ࢆ, Cell Counting Kit-8 (ᰴᘧ఍♫ ྠோ໬Ꮫ◊✲ᡤ) ࢆ⏝࠸ࡓ⏕⣽⬊ᩘࡢィ ࡟ࡼࡾ
☜ㄆࡋࡓ (Fig. 3)ࠋ௨ୖࡢ⤖ᯝ࠿ࡽ, NASH ࣔࢹࣝ⣽⬊ࡢస〇ࡀ࡞ࡉࢀࡓࡇ࡜ࡀ☜ㄆࡉࢀࡓࡓ
ࡵ, ࡇࡢ⣽⬊⣔ࢆ௨ᚋࡢᐇ㦂࡟౑⏝ࡍࡿࡇ࡜࡜ࡋࡓࠋ



Fig. 1. Intracellular triglyceride.
HuH-7 cells were untreated or treated 600 PM FAs for 24
hours, at a molar ratio of 1: 2 (palmitic and oleic acid,
respectively). Increase of intracellular fat accumulation was
determined by TG assay kit. Data expressed mean ±
standard deviation, n=3. **P < 0.01 vs FA (-). FA (-),
untreated with FA; FA (+), treated with FA.

Ө
Fig. 2. IL-8 release from the HuH-7 cells.
Culture supernatants were harvested at 24 h after untreated
or treated with FAs, and IL-8 levels were measured by
ELISA. Data expressed mean ± standard deviation, n=3.
**P < 0.01 vs FA (-). FA (-), untreated with FA; FA (+),
treated with FA.

Ө

Fig. 3. Cell viability assay.
The effect of the FA on the viability of the HuH-7 cells.
Cells were treated with FAs for 24 h and assessed with Cell
Counting Kit-8. Cells treated with vehicle alone served as a
control. Data expressed mean ± standard deviation, n=3. FA
(-), untreated with FA; FA (+), treated with FA.



➨஧⠇ NASH ࣔࢹࣝ⣽⬊ࢆ⏝࠸࡚ࡢⓎ㓝⏕⸆ᢳฟ≀ࡢᢠ⅖⑕స⏝
SN13T ᰴࡀᗄࡘ࠿ࡢ⏕⸆Ỉᢳฟᾮ୰࡛Ⰻዲ࡟ቑṪ࡛ࡁࡓࡓࡵ, ࡇࡢᰴࢆ⏝࠸࡚ Table 1 ࡟♧
ࡋࡓ⏕⸆ࢆⓎ㓝ࡉࡏ, Ⓨ㓝⏕⸆ᢳฟ≀ࢆస〇ࡋࡓࠋḟ࡟, NASH ࣔࢹࣝ⣽⬊ࢆ⏝࠸࡚, Ⓨ㓝⏕
⸆ᢳฟ≀࡟࠾ࡅࡿᢠ⅖⑕άᛶࢆࢫࢡ࣮ࣜࢽࣥࢢࡋࡓࠋࡑࡢ⤖ᯝ, Ⱅⴥ (࢞࢖ࣚ࢘) Ⓨ㓝≀࡟࠾
࠸࡚᭷ព࡞ᢠ⅖⑕άᛶࡀ☜ㄆࡉࢀࡓ (Fig. 4)ࠋ

Table 1. Medicinal herbs used for screening.



Fig 4. The inhibitory effect of the herb extract fermented with SN13T strain on the release of IL-8 from HuH-7
cells (a1, a2) and cell viability assay after treatment (b1, b2).
The herb extracts were incubated for 24 h at 28℃ with SN13T strain. The resulting samples were added to the HuH-7
cell culture medium at 50 Pg/mL, along with FA treatment, except for P. frutescens, and C. cassia (20 Pg/mL). The
culture supernatant of the cells was collected 24 h after the treatment. The amounts of IL-8 were measured by the ELISA
method. Data are expressed as the mean ± standard deviation, (n=2–3). **P < 0.01 vs. control.



➨୕⠇ ␗࡞ࡿ᳜≀ங㓟⳦࡛Ⓨ㓝ࡉࡏࡓⰕⴥᢳฟ≀ࡢᢠ⅖⑕స⏝
NASH ࣔࢹࣝ⣽⬊ࢆ⏝࠸ࡓᢠ⅖⑕άᛶࢫࢡ࣮ࣜࢽࣥࢢ࡟࠾࠸࡚, Ⓨ㓝Ⱅⴥᢳฟ≀࡟άᛶࡀ
ㄆࡵࡽࢀࡓࡓࡵ, ᮡᒣ◊✲ᐊ࡟࠾࠸࡚඲ࢤࣀ࣒ゎㄞࡀ࡞ࡉࢀ࡚࠸ࡿ᳜≀ங㓟⳦ᰴ (Lb.
plantarum SN13T, Lb. plantarum SN35N, Pediococcus (P.) pentosaceus LP28) ࢆ㑅ᢥࡋ, ྛᰴ࡛Ⓨ
㓝ࡉࡏࡓⰕⴥᢳฟ≀ࡢຠᯝࢆᮍⓎ㓝ࡢⰕⴥᢳฟ≀࡜ẚ㍑ࡋࡓࠋྛங㓟⳦ᰴࢆⰕⴥỈᢳฟᾮ࡟
᳜⳦ࡋ, 28℃࡛ 24 ᫬㛫㟼⨨ᇵ㣴ࡋࡓࠋ㐲ᚰ᧯స࡟ࡼࡾ, ࡑࡢᇵ㣴ᾮ࠿ࡽ⳦యࢆ㝖ࡁ, ᚓࡽࢀࡓ
ୖΎࢆⓎ㓝ࢧࣥࣉࣝ࡜ࡋࡓࠋங㓟⳦ᇵ㣴↓ࡋࡢࢧࣥࣉࣝࡶྠ᫬ㄪ〇ࡋ, ࡑࢀࢆᮍⓎ㓝ࢧࣥࣉ
ࣝ࡜ࡋࡓࠋᇵ㣴ᚋࡣ, ࡝ࡢᰴࡢᇵ㣴ୖΎࡶྠ⛬ᗘ pH ࡀపୗࡋ, Ⓨ㓝ࡀ࡞ࡉࢀ࡚࠸ࡿࡇ࡜ࡀ☜
ㄆࡉࢀࡓ (Table 2)ࠋ
ࡇࡢࡼ࠺࡟ࡋ࡚ᚓࡽࢀࡓⓎ㓝Ⱅⴥᢳฟ≀࠾ࡼࡧᮍⓎ㓝Ⱅⴥᢳฟ≀ࡢᢠ⅖⑕άᛶࢆ, HuH-7
⣽⬊ࢆ⏝࠸ࡓ NASH ࣔࢹࣝ⣽⬊࠿ࡽ㐟㞳ࡉࢀࡿ IL-8 㔞ࢆᣦᶆ࡜ࡋ᳨࡚ドࡋࡓࠋᮍⓎ㓝ࡢⰕⴥ
ᢳฟ≀ (u-AP) ࡣ, FA ฎ⌮ࡋࡓ HuH-7 ⣽⬊࠿ࡽ㐟㞳ࡉࢀࡓ IL-8 㔞ࢆ 50%࡟ῶᑡࡉࡏࡓ (Fig.
5a)ࠋࡉࡽ࡟, SN13T ᰴ (f-AP-13T) ࡲࡓࡣ SN35N ᰴ (f-AP-35N) ࡛Ⓨ㓝ࡋࡓⰕⴥᢳฟ≀ࡣ, ᮍ
Ⓨ㓝ᢳฟ≀࡜ẚ㍑ࡋ࡚㐟㞳 IL-8 㔞ࢆࡉࡽ࡟ῶᑡࡉࡏࡓࠋຍ࠼࡚, SN13T ᰴ࡛Ⓨ㓝ࡉࡏࡓⰕⴥ
ᢳฟ≀ࡢ IL-8 ᢚไຠᯝࡣ, SN35N ᰴ࡛Ⓨ㓝ࡉࡏࡓሙྜࡼࡾࡶ㧗࠿ࡗࡓࠋ௚᪉, LP28 ᰴ࡛Ⓨ㓝
ࡉࡏࡓⰕⴥᢳฟ≀ (f-AP-LP28) ࡣ, ᮍⓎ㓝ࡢⰕⴥᢳฟ≀࡜࡯ࡰྠࡌ⛬ᗘ࡟ᢚไࡉࢀࡓࠋ
௨ୖࡢ⤖ᯝ࠿ࡽ, ≉ᐃࡢங㓟⳦ᰴ࡟ࡼࡗ࡚Ⓨ㓝ࡉࡏࡓⰕⴥᢳฟ≀࡟࠾࠸࡚ᢠ⅖⑕άᛶࡀቑ
ᙉࡍࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡾ, Lb. plantarum SN13T ࡣ, Ⱅⴥࡢᢳฟ≀ࢆⓎ㓝ࡉࡏࡿୖ࡛, 3 ࡘࡢ
⳦ᰴࡢ୰࡛᭱ࡶ㐺ࡍࡿࡇ࡜ࡀ♧ࡉࢀࡓࠋ࡞࠾, ࡇࡢᐇ㦂࡛⏝࠸ࡓྛ⃰ᗘࡢⰕⴥᢳฟ≀࠾ࡼࡧ
FA ࡣ, HuH-7 ⣽⬊࡟ࡣ⣽⬊ẘᛶࢆ♧ࡉ࡞࠸ࡇ࡜ࡶ, Cell Counting Kit-8 ࢆ⏝࠸ࡓ⏕⣽⬊ィ ࢔
ࢵࢭ࢖࡟ࡼࡾ☜ㄆࡋࡓ (Fig. 5b)ࠋ
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Table 2. pH of AP extract after fermentation with each LAB strain.

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Fig. 5. The inhibitory effect of the AP extract fermented with each LAB strain on the release of IL-8 from HuH-7
cells (a) and cell viability assay after treatment (b).
In this experiment, Lb. plantarum SN13T, Lb. plantarum SN35N, and P. pentosaceus LP28 were used. ...

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