Toll-like receptor(TLR)࡟ࡼࡾㄆ㆑ࡉࢀࠊ⅖⑕࡟㛵ࢃࡿ㑇ఏᏊⓎ⌧ࢆㄏᑟࡍ

ㄽᩥࡢෆᐜࡢせ᪨

Ặྡ㸸Ụ ⸨ ᛅ ὒ

ᑓᨷศ㔝ࡢྡ⛠㸸༤ኈ㸦⸆Ꮫ㸧

ㄽᩥ㢟ྡ㸸ኳ↛≀࠿ࡽࡢᢠ⅖⑕≀㉁࠾ࡼࡧࡀࢇ⸆๣⪏ᛶඞ᭹≀㉁ࡢ᥈⣴

ᵝࠎ࡞ᢠ⏕≀㉁࠿ࡽጞࡲࡾࠊ࣐࢖ࢺ࣐࢖ࢩࣥࠊࢱ࢟ࢯ࣮ࣝࠊࣅࣥࢡࣜࢫࢳࣥ࡞࡝ኳ↛⏤᮶ࡢ⏕≀άᛶ≀

㉁ࡣࡇࢀࡲ࡛࡟ᩘከࡃぢ࠸ࡔࡉࢀ࡚࠾ࡾࠊࡑࢀࡽࡣ་⸆ရ࡜ࡋ࡚࠶ࡿ࠸ࡣ་⸆ရࡢ࣮ࣜࢻ໬ྜ≀࡜ࡋ࡚౪ ࡉࢀࡓࠋ㏆ᖺࠊከᵝ࡞໬ྜ≀ࡢྜᡂࡀྍ⬟࡜࡞ࡾேᕤⓗ࡞࣮ࣜࢻ໬ྜ≀ࡢ᥈⣴ࡀ୺ὶ࡜࡞ࡾࡘࡘ࠶ࡿࠋࡋ

࠿ࡋ࡞ࡀࡽࠊኳ↛≀࠿ࡽࡢ໬ྜ≀᥈⣴ࡣ࣓ࣂࣟࢳࣥࠊࢱࢡ࣒ࣟࣜࢫ㸦FK506㸧ࡢᵝ࡞᪂つ㦵᱁ࢆᣢࡘ᪂ࡓ

࡞໬ྜ≀ࡢⓎぢࡸ᪂ࡓ࡞స⏝ࢆᣢࡕᮍࡔ἞⒪ἲࡢ☜❧ࡋ࡚࠸࡞࠸⑌ᝈ➼࡟౪ࡍࡿࡇ࡜ࡢ࡛ࡁࡿ໬ྜ≀Ⓨぢ ࡢྍ⬟ᛶࢆ⛎ࡵ࡚࠸ࡿࠋᮏ◊✲ࡣ

NO

ࡼࡧ⸆๣⪏ᛶ⒴࡟ᑐࡍࡿ⪏ᛶඞ᭹స⏝ࢆᣦᶆ࡜ࡋ࡚ࠊ᪂ࡓ࡞⪏ᛶඞ᭹స⏝ࢆ᭷ࡍࡿ໬ྜ≀ࡢ᥈⣴ࢆ┠ⓗ࡜

ࡋ࡚⾜ࡗࡓࠋ

ࠑࠑNO⏘⏕ᢚไࢆᣦᶆ࡜ࡋࡓᢠ⅖⑕≀㉁ࡢ᥈⣴ࠒ

⑓ཎయ࡟≉␗ⓗ࡞ᵓᡂᡂศࡣ

Toll-like receptor(TLR)࡟ࡼࡾㄆ㆑ࡉࢀࠊ⅖⑕࡟㛵ࢃࡿ㑇ఏᏊⓎ⌧ࢆㄏᑟࡍ

ࡿࠋTLRࡣࣄࢺ࡛ࡣ 10 ✀㢮ࡢ ࣐࢘ࢫ࡛ࡣ

12

ศᏊ࡟ࡼࡾ␗࡞ࡿࢩࢢࢼࣝఏ㐩⤒㊰㸦Fig.1㸧ࢆάᛶ

໬ࡋࠊ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥࠊࢣࣔ࢝࢖ࣥࡸ࢖ࣥࢱ࣮

ࣇ࢙ࣟࣥࡢ⏘⏕ࢆᘬࡁ㉳ࡇࡍ⤖ᯝࠊ⾑⟶㏱㐣ᛶࡢஹ 㐍ࠊⓑ⾑⌫ࠊ࣐ࢡࣟࣇ࢓࣮ࢪࡢ㐟㉮ࡸάᛶ໬ࡀ㉳ࡇ

ࡾࠊឤᰁᚤ⏕≀ࡢ᤼㝖ࢆಁࡍࠋ㏆ᖺ࡛ࡣࡇࢀࡽ

TLR

࡞᪂つࢩࢢࢼࣝఏ㐩≀㉁ࡢⓎぢࡸࡑࡢᶵ⬟ゎᯒࡀ⾜

ࢃࢀ࡚࠸ࡿࠋ

TLR

ࡎࠊ⋓ᚓච␿⣔ࡢάᛶ໬࡟ࡶ㔜せ࡞ാࡁࢆᢸࡗ࡚࠸

ࡿࡇ࡜ࡀ᫂ࡽ࠿࡟࡞ࡗ࡚࠸ࡿࠋࡋ࠿ࡋ࡞ࡀࡽ

TRL

ࢆ௓ࡍࡿච␿ᛂ⟅ࡣ✀ࠎࡢච␿⑌ᝈ㸦࢚ࣥࢻࢺ࢟ࢩ

ࣥࢩࣙࢵࢡࠊ៏ᛶ⅖⑕ᛶ⭠⑌ᝈࠊ඲㌟ᛶ࢚ࣜࢸ࣐ࢺ

࣮ࢹࢫ ➼㸧ࡢⓎ⑕࡟ࡶ῝ࡃ㛵୚ࡋ࡚࠸ࡿ࡜⪃࠼ࡽࢀ

࡚࠾ࡾࠊࡇࡢࡼ࠺࡞ච␿⣔⑌ᝈࢆඞ᭹ࡍࡿࡓࡵ࡟ࡣ

TLR

ࡑࡇ࡛ᮏ◊✲࡛ࡣࡑࡢ⏘⏕࡟

TLR

㸦NO㸧࡟╔┠ࡋࠊ࣐࢘ࢫ࣐ࢡࣟࣇ࢓࣮ࢪᵝ⣽⬊ࢆ⏝࠸ᚤ⏕≀஧ḟ௦ㅰ⏘≀࠾ࡼࡧ᳜≀ᢳฟ≀࠿ࡽ

lipopolysaccharide㸦LPS㸧่⃭࡟ࡼࡿ NO

1.

NO

ࢫࢡ࣮ࣜࢽࣥࢢ⣔࡟࠾࠸࡚ࡣࠊ࣐࢘ࢫ࣐ࢡࣟࣇ࢓࣮ࢪᵝ⣽⬊ᰴ࡛࠶ࡿ

RAW264.7

ᇵ㣴⏬ศᏑᅾୗ࡛ ᫬㛫๓ฎ⌮ࡋࠊ/36㸦ǍJP/㸧ࢆῧຍࡋ ᫬㛫ࡢᇵ㣴ᚋࠊᇵ㣴ᾮ୰ࡢ 12 ⃰ᗘࢆ *ULHVV ἲࢆ⏝࠸࡚ ᐃࡋࡓࠋ⣒≧⳦ࡢ ᇵ㣴⏬ศ࡛ࢫࢡ࣮ࣜࢽࣥࢢࢆ⾜ࡗࡓ⤖ᯝࠊ

3HQLFLOOLXP

,08 ࡟᭱ࡶᙉ࠸12 ⏘⏕ᢚไస⏝ࢆぢฟࡋࡓࠋᮏ⳦ᰴࢆ⡿ࢆ୺ᡂศ࡜ࡍࡿ⏕⏘ᇵᆅ࡛᪥㛫㟼⨨ᇵ 㣴ࡋࠊ㓑㓟࢚ࢳࣝࢆ⏝࠸࡚ᢳฟᚋࠊᚓࡽࢀࡓᢳฟ⏬ศࢆࢩࣜ࢝ࢤ࣒ࣝ࢝ࣛࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮࡛ศ㞳ࡋࠊ 2'6࣒࢝ࣛศྲྀ+3/&࡟ࡼࡾࡘࡢάᛶᡂศ EHUNHOH\DFHWDO$ࠊ%ࠊ& ࢆ༢㞳ࡋࡓࠋࡇࢀࡽ໬ྜ≀ࡢෆࠊ

᭱ࡶᙉ࠸άᛶࢆ♧ࡋࡓ EHUNHOH\DFHWDO&㸦%$&㸧ࢆ⏝࠸ᢠ⅖⑕స⏝࣓࢝ࢽࢬ࣒ࡢ᳨ウࢆ⾜ࡗࡓࠋ

2. Berkeleyacetal C㸦BAC㸧ࡢᢠ⅖⑕స⏝స⏝࣓࢝ࢽࢬ࣒ゎᯒ

RAW264.7

LPS

peptidoglycan(PGN)่⃭࡟ᑐࡍࡿ BAC

NO

PGE2ࠊ TNF-ǂࠊ iNOS

COX-2

NO

Griess

PGE2ࠊ TNF-ǂ

࠾࠸࡚ࡣ

ELISA

Western blot

Fig.1 Toll-like receptor signaling pathway

BAC

TLR

ࡍࡿࡇ࡜ࢆ♧၀ࡋ࡚࠸ࡿࠋTLRࢩࢢࢼࣝఏ㐩࡟࠾ࡅࡿ

MyD88

ࡢ

BAC

Western blot

NF-NjB

Ikk-ǂǃࠊ

TAK1

Aktࠊ JNKࠊ P38MAPKࠊ 30$3.

3.

IRAK-1

BAC

IRAK-1

⤒㊰࡟࠾࠸࡚ࡑࡢୖὶ࡟Ꮡᅾࡍࡿ≀㉁࡛࠶ࡿࡇ࡜

ࡀ♧၀ࡉࢀࡓࠋ

TLR

IRAK-1

IRAK-4

kinase activity

BAC

IRAK-4

kinase activity

ศᏊࡣ

IRAK-4

୍᪉ࠊ

LPS

TLR

IRAK-4

MyD88

IRF-3

BAC

ࡇࢀࡽࡢ⤖ᯝ࠿ࡽࠊBACࡢᢠ⅖⑕స⏝࡟࠾ࡅࡿᶆⓗศᏊࡣ

IRAK-4

IRAK-4

TLR

࠸ࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗ࡚࠸ࡿࠋࡲࡓ

BAC

ྠᵝࡢస⏝ࢆ♧ࡋࡓࠋTLRࡢ⅖⑕཯ᛂ࡟࠾ࡅࡿᙺ๭ࢆ♧ࡍࡇ࡜࠿ࡽ

BAC

⸆ࡢ࣮ࣜࢻ໬ྜ≀࡜࡞ࡿྍ⬟ᛶࡀ࠶ࡿࡶࡢ࡜⪃࠼ࡽࢀࡿࠋ

3. erythraline

ᚤ⏕≀஧ḟ௦ㅰ⏘≀ࢆ⏝࠸࡚⾜ࡗࡓࢫࢡ࣮ࣜࢽࣥࢢ࡜ྠᵝ࡟

LPS

NO

࡛࠶ࡿ

Erythrina crista-galli

erythralineࠊerythrinineࠊhypaphorine

3

NO

ǍJP/ ࡛࠶ࡗࡓࠋࡑࢀࡽ໬ྜ≀ࡢෆࠊస⏝࣓࢝ࢽࢬ࣒ࡢ᳨ウࢆ⾜࠺ୖ࡛༑ศ࡞཰㔞ࢆᚓࡿࡇ࡜ࡀฟ᮶ࡓ erythraline

LPS

ࡿ

NO

iNOS

2. TLR

LPS

NF-NjB

3. INjB-ǂ

degradation

Ikk-ǂǃࠊࡢࣜࣥ㓟໬ࡢᢚไࡀㄆࡵࡽࢀࡓࠋ୍᪉ࠊ

TAK1ࡢࣜࣥ㓟໬࠾ࡼࡧ IRAK-1ࡢศゎ࡟ࡘ࠸࡚ࡣᢚ

TAK1

INjB-ǂ

Ikk-ǂǃ

JNK

P38MAPK

♧ࡉࢀࡓࠋࡇࢀࡽࡢ⤖ᯝࡣ

PGN㸦TLR2㸧่⃭࡟ࡼࡿࢩࢢࢼࣝఏ㐩࡟࠾࠸࡚ࡶྠᵝࡢ⤖ᯝ࡛࠶ࡗࡓࠋ

erythraline

TAK1

TLR

IL-1

erythraline

E. crista-galli

TAK1 kinase activity

၀ࡉࢀࡓࠋ

ࠑኳ↛≀࠿ࡽࡢࡀࢇ⸆๣⪏ᛶඞ᭹≀㉁ࡢ᥈⣴ࠒ

ࡀࢇࡣ⌧ᅾ᪥ᮏ࡟࠾࠸࡚Ṛஸཎᅉࡢ➨୍఩࡜࡞ࡗ࡚࠸ࡿ஦ࡀ▱ࡽࢀ࡚࠸ࡿࠋ௒᪥࡟࠾ࡅࡿࡀࢇࡢ἞⒪ἲ ࡣ୺࡟እ⛉ᡭ⾡࡟ࡼࡿᝈ㒊ࡢ㝖ཤࠊᢠ⭘⒆⸆࡟ࡼࡿ໬Ꮫ⒪ἲࠊᨺᑕ⥺ࡢ↷ᑕ࡟ࡼࡿᨺᑕ⥺⒪ἲ࡛࠶ࡿࠋࡇ

ࢀࡽ἞⒪ἲࡢ୰࡛ࠊ໬Ꮫ⒪ἲࡣᵝࠎ࡞ไࡀࢇ๣ࡢ㛤Ⓨ࡟ࡼࡾ᭷ຠ࡞ࡀࢇࡢ἞⒪ἲ࡜ࡋ࡚ὀ┠ࡉࢀࡿࡼ࠺࡟

࡞ࡗࡓࠋ≉࡟ⓑ⾑⑓࡟௦⾲ࡉࢀࡿ⾑ᾮࡢࡀࢇ࠾࠸࡚໬Ꮫ⒪ἲࡣࠊᨺᑕ⥺⒪ἲ࡜୪ࢇ࡛ࠊ᭱ࡶ᭷ᮃ࡞἞⒪ἲ

࡜࡞ࡗ࡚࠸ࡿࠋࡋ࠿ࡋ࡞ࡀࡽࡀࢇࡢ἞⒪࡟ᢠ⭘⒆⸆ࡀᗈࡃ㐺⏝ࡉࢀࡿ஦࡛ᢠ⭘⒆⸆࡟ᑐࡍࡿࡀࢇ⣽⬊ࡢ⪏

Fig.2 Effect of BAC on the kinase activity of IRAK-4

ไࡀࢇ๣࡟ᑐࡋ࡚ࡶࠊ⪏ᛶࢆᣢࡕከ๣⪏ᛶ໬ࡍࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋᵝࠎ࡞⸆๣࡟ᑐࡍࡿ⪏ᛶࡣࠊP㸫⢾

ࢱࣥࣃࢡ㉁ࡢ⸆๣᤼ฟࡀཎᅉࡢ୍ࡘ࡜ࡉࢀ࡚࠸ࡿࠋࡇࡢ

P㸫⢾ࢱࣥࣃࢡ㉁࡟ไࡀࢇ๣ࡢ௦ࢃࡾ࡟⤖ྜࡋࠊ

ࢀࡿࠋ⌧ᅾࠊ⸆≀⪏ᛶ⒴ඞ᭹≀㉁ࡢⓎぢࡸඞ᭹๣ࡢ㛤Ⓨࡣࡀࢇ἞⒪ࡢୖ࡛㔜せ࡞ㄢ㢟࡛࠶ࡿࡀࠊᮍࡔ⮫ᗋ

࡟౪⤥ࡉࢀ࡚࠸ࡿ⸆≀ࡣᏑᅾࡋ࡞࠸ࠋࡑࡇ࡛ᮏ◊✲࡛ࡣኳ↛≀࠿ࡽࡢࡀࢇ⸆≀⪏ᛶඞ᭹స⏝ࢆᣢࡘ໬ྜ≀

ࡢ᥈⣴ࢆ┠ⓗ࡜ࡋࡓࠋ

ࢫࢡ࣮ࣜࢽࣥࢢ⣔࡟ࡣࣅࣥࢡࣜࢫࢳࣥ㸦VCR㸧ឤཷᛶᡥᖹୖ⓶⒴⣽⬊ᰴ࡛࠶ࡿ

.%6

ࣜࢫࢳࣥ⪏ᛶ⒴⣽⬊ᰴ࡛࠶ࡿ

.%9-

.%9-

ࣉࣝࢆῧຍࡋ

72

VCR

VCR

ࡇࡢ᪉ἲࢆ⏝࠸࡚ኳ↛≀ࡼࡾࡀࢇ⸆≀⪏ᛶඞ᭹≀㉁ࡢ᥈⣴ࢆ⾜ࡗࡓ࡜ࡇࢁࠊ࣐࣮ࣞࢩ࢔⏘᳜≀ᡂศ࠿ࡽ

ᩘ✀ࡢ᪂つ

indole alkaloids

.%6

࠸࡚ࣅࣥࢡࣜࢫࢳࣥࡢ ,&್ࡢ

QJP/

25ǍJP/ ῧຍࡢ᮲௳࡟࠾࠸࡚ࡶ⣽⬊ẘᛶࢆ♧ࡉ࡞

࠸ࡇ࡜ࡀ☜ㄆࡉࢀࠊࡲࡓ༢⊂࡛ࡢస⏝࡟࠾

࠸࡚ࡣ⪏ᛶᰴ࡛࠶ࡿ

.%9-

࡚ࡶ⣽⬊ẘᛶࢆ♧ࡉ࡞࠸஦ࡀ☜ㄆࡉࢀࡓࠋ ࡑࡢ᮲௳ୗ࡛ࡇࢀࡽ໬ྜ≀ࡣ

VCR

⏝࡟࠾࠸࡚⣽⬊ẘᛶࢆ♧ࡋࡓࠋ௦⾲ⓗ࡞໬

ྜ≀ࡢ⸆≀⪏ᛶඞ᭹࡟࠾ࡅࡿ ,&ࢆ⾲࡟♧

ࡍࠋ௨ୖࡢ⤖ᯝࡼࡾࠊࡇࢀࡽ໬ྜ≀ࡣ⸆≀

⪏ᛶࡀࢇ࡟ᑐࡍࡿ⪏ᛶඞ᭹స⏝ࢆᣢࡘ⸆๣

ࡢ࣮ࣜࢻ໬ྜ≀࡜࡞ࡾ࠺ࡿࡇ࡜ࡀ♧၀ࡉࢀ

ࡓࠋ

௒ᚋࠊᮏ◊✲࡟ࡼࡾぢฟࡉࢀࡓ໬ྜ≀ࢆ⏝࠸ࡓ᭦࡞ࡿస⏝ᶵᗎࡢゎ᫂ࡸࠊ

LQYLYR

᳨ウࡉࢀࡿࡇ࡜࡟ࡼࡾࠊᢠ⅖⑕࠾ࡼࡧࡀࢇ⸆๣⪏ᛶඞ᭹స⏝ࢆᣢࡘ⸆≀ࡢ㛤Ⓨ࡟⤖ࡧ௜ࡃྍ⬟ᛶࡀ㧗࠸࡜

⪃࠼ࡽࢀࡿࠋ