࠾࠸࡚ࡣ㸪LPSㄏⓎᛶࡢPGE2⏘⏕ࡣinhibitorฎ⌮ࡋ࡞ࡗࡓ⣽⬊ (Fig. 5) ẚ㍑ࡋ࡚1/5⛬ᗘᢚไࡉࢀࡓࠋ࡞࠾㸪ࡇࢀࡽLPSㄏⓎᛶࡢPGE2⏘⏕ࡣ㸪HT
ࡼࡗ࡚⃰ᗘ౫ᏑⓗᢚไࡉࢀࡓࠋCelastrol࡛ฎ⌮ࡋࡓ⣽⬊࠾࠸࡚ࡣ㸪LPSㄏⓎ
ᛶࡢPGE2⏘⏕ࡢኚࡣㄆࡵࡽࢀ࡞ࡗࡓ (Fig. 9)ࠋ
2. RAW264.7⣽⬊࠾ࡅࡿNF-κB㛵㐃ࢱࣥࣃࢡ㉁ࡢⓎ⌧
29
Western blottingἲࢆ⏝࠸࡚㸪ྛinhibitor࡛ฎ⌮ࡋࡓRAW264.7⣽⬊ࡘ࠸࡚㸪 HTᏑᅾࡲࡓࡣ㠀Ꮡᅾୗ࡛ࡢpNF-κBཬࡧpERK1/2ࡢࢱࣥࣃࢡ㉁Ⓨ⌧ࢆホ౯ࡋࡓࠋ Celastrolฎ⌮⣽⬊࡛ࡣ㸪LPS (0.25 μg/mL) ࡢῧຍࡼࡗ࡚ࡶ㸪pNF-κBࡢⓎ⌧ࡣ
ࡃㄆࡵࡽࢀ࡞ࡗࡓ (Fig. 10A)ࠋU0126ฎ⌮⣽⬊࡛ࡣ㸪inhibitorฎ⌮ࡋ࡞ࡗࡓ⣽
⬊ (Fig. 6B) ẚ㍑ࡋ࡚㸪pERK1/2ࡢⓎ⌧ࡀ᫂░࡛࠶ࡗࡓࠋࡲࡓ㸪pERK1/2ࡢ
Ⓨ⌧ࡣ㸪HTࡢᏑᅾࡼࡗ࡚ᢚไࡉࢀ࡞ࡗࡓ (Fig. 10B)ࠋp38ཬࡧJNK㛵ࡋ࡚
ࡣLPS่⃭ࡼࡿࣜࣥ㓟ࡀ☜ㄆ࡛ࡁ࡞ࡗࡓࡓࡵ㸪inhibitorࢆ⏝࠸ࡓᐇ㦂ࡣᐇ
ࡋ࡞ࡗࡓ (ࢹ࣮ࢱ㠀⾲♧)ࠋ
30
⪃ᐹ
HTࡀస⏝ࡍࡿࢩࢢࢼࣝఏ㐩⤒㊰ࢆ᫂ࡽࡍࡿࡓࡵ㸪NF-κBཬࡧERK1/2㸪p38㸪 JNKࡢ3ࡘMAPKᑐࡋ࡚㸪inhibitorࢆ⏝࠸࡚ྛ⤒㊰ࢆ㜼ᐖࡋࡓ㝿ࡢHTࡢస⏝
ࡢኚࢆホ౯ࡋࡓࠋ
NF-κB㜼ᐖ⸆࡛࠶ࡿcelastrolࡣRAW264.7⣽⬊࠾ࡅࡿNOࡢ⏘⏕ࢆᢚ ไࡋࡓࡀ㸪ERK1/2㜼ᐖ⸆࡛࠶ࡿU0126㸪p38㜼ᐖ⸆࡛࠶ࡿSB203580㸪JNK㜼ᐖ
⸆࡛࠶ࡿSP600125ࡘ࠸࡚ࡣ㸪NO⏘⏕ࡢᢚไస⏝ࡣㄆࡵࡽࢀ࡞ࡗࡓࠋLPSㄏ
ⓎᛶࡢNO⏘⏕ྛMAPKࡢ㛵ಀࡣࡇࢀࡲ࡛」ᩘሗ࿌ࡉࢀ࡚࠸ࡿࠋChanࡽ (32) ࡣ㸪ERK1/2㜼ᐖ⸆࡛࠶ࡿPD-98059ཬࡧp38㜼ᐖ⸆࡛࠶ࡿSB-203580ࢆRAW264.7
⣽⬊ฎ⌮ᚋ LPS่⃭ࡋࡓ⤖ᯝ㸪PD-98059 ฎ⌮⣽⬊࡛ࡣ NO ࡢ⏘⏕ࡀᢚไࡉ
ࢀࡓࡀ㸪SB203580 ฎ⌮⣽⬊࡛ࡣᢚไࡣㄆࡵࡽࢀ࡞ࡗࡓሗ࿌ࡋ࡚࠸ࡿࠋ୍᪉
࡛㸪Bhatࡽ (50) ࡣࢢࣜ⣽⬊ࢆ⏝࠸ࡓᐇ㦂ࢆ⾜࠸㸪ERK1/2㸪p38࠸ࡎࢀࡢ⤒㊰
ࢆ㜼ᐖࡋࡓሙྜ࡛ࡶ㸪LPSㄏⓎᛶࡢNOࡣᢚไࡉࢀࡓሗ࿌ࡋ࡚࠸ࡿࠋࡲࡓ㸪Chen
ࡽ (51) ࡣLPSㄏⓎᛶࡢiNOSࡢୖ᪼ࡣp38⤒㊰ࡼࡗ࡚ไᚚࡉࢀࡿࡀ㸪ERK1/2
⤒㊰ࡣ㛵ࡋ࡞࠸ሗ࿌ࡋ࡚࠸ࡿࠋJNK⤒㊰㛵ࡋ࡚ࡶ㸪LPSㄏⓎᛶࡢNO⏘⏕
ಁ㐍ⓗാࡃ࠸࠺ሗ࿌ࡶ࠶ࡿ୍᪉࡛ (52)㸪JNK⤒㊰ࢆ㜼ᐖࡋ࡚ࡶNO⏘⏕
ኚࡣ࡞࠸࠸࠺ሗ࿌ࡶ࠶ࡿ (32)ࠋᅇࡢᐇ㦂࡛ࡣ㸪NF-κB㜼ᐖ⸆࡛࠶ࡿcelastrol ௨እࡣNO⏘⏕┤᥋ᙳ㡪ࢆ࠼࡞ࡗࡓࠋᮏ⤖ᯝࡼࡾ㸪ྛMAPKࡣLPS (0.25 μg/mL) ่⃭᮲௳ୗࡢ RAW264.7 ⣽⬊࠾࠸࡚ࡣ༢⊂࡛ࡣ NO ࡢ⏘⏕ࢆಁ㐍ࡍࡿ
ࡇࡣ࡞ࡃ㸪NF-κBࡢάᛶࡀNOࡢ⏘⏕యⓗ㛵ࢃࡗ࡚࠸ࡿྍ⬟ᛶࡀ♧၀ ࡉࢀࡓࠋᮏ⤖ᯝࡣ㸪ྛMAPK ࡀNF-κB ࡢάᛶ⿵㊊ⓗാࡃ࠸࠺◊✲ሗ࿌
31
(53) ୍⮴ࡍࡿࡶࡢ࡛࠶ࡿࠋࡲࡓ㸪p38ཬࡧJNK⤒㊰࡛ࡣactivator protein 1 (AP-1) ࡢάᛶࢆࡋ࡚iNOSࡢ⏘⏕ࢆቑຍࡉࡏࡿࡀ㸪AP-1ࡢάᛶࡣLPS่⃭ࡼࡗ࡚
㉳ࡇࡽ࡞࠸࠸࠺ሗ࿌ࡶ࠶ࡿ (54, 55)ࠋࡇࡢሗ࿌ࡣ㸪ᅇWestern blotting࠾࠸
࡚LPS ่⃭ࡼࡿp38 ཬࡧJNK ࡢࣜࣥ㓟ࢆᤊ࠼ࡽࢀ࡞ࡗࡓᐇࢆࡅࡿ
ࡶࡢ⪃࠼ࡽࢀࡿࠋ
HTࡣྛMAPKࡢ㜼ᐖࡢ᭷↓㛵ࢃࡽࡎ㸪⃰ᗘ౫ᏑⓗNO⏘⏕ࢆᢚไࡋࡓࡇ
ࡽ㸪HTࡢస⏝ⅬࡣERK1/2㸪p38㸪JNK⤒㊰ୖࡣ↓ࡃ㸪ࡘNF-κB⤒㊰ࡼ
ࡾࡶୗὶ࠶ࡿྍ⬟ᛶࡀ㧗࠸⪃࠼ࡽࢀࡓࠋ➨ 1 ❶ࡢ⤖ᯝࢆ㋃ࡲ࠼ࡿ㸪HT ࡣ 12.5 μg/mLࡢ⃰ᗘ࡛ࡣNF-κBࢩࢢࢼࣝఏ㐩⤒㊰ࢆᢚไࡍࡿࡇ࡛㸪iNOS㸪COX-2
ཬࡧIL-1β㑇ఏᏊࡢⓎ⌧ࢆᢚไࡋ㸪12.5 μg/mLࡼࡾప࠸⃰ᗘ࡛ࡣNF-κB⤒㊰ࡼࡾ
ࡶୗὶ࠶ࡿࢩࢢࢼࣝ⤒㊰స⏝ࡋ࡚㸪ୖグࡢ㑇ఏᏊ⩌ࢆᢚไࡋ࡚࠸ࡿ⪃࠼ࡽ
ࢀࡿࠋ
TNF-α㛵ࡋ࡚㸪U0126 (ERK1/2⤒㊰㜼ᐖ)㸪SB203580 (p38㜼ᐖ) ཬࡧSP600125 (JNK 㜼ᐖ) ࡛ฎ⌮ࡋࡓ⣽⬊࡛ࡣ㸪LPSㄏⓎᛶࡢ TNF-α ⏘⏕ࡣ inhibitor ฎ⌮ࡋ࡞
ࡗࡓ⣽⬊ẚ㍑ࡋ࡚ 1/5ࠥ1/10 ⛬ᗘᢚไࡉࢀࡓࠋᮏ⤖ᯝࡣ㸪ྠᵝ inhibitor
ࢆ⏝࠸ࡓ㐣ཤࡢሗ࿌ࡶ୍⮴ࡋ࡚࠾ࡾ㸪ERK1/2ཬࡧJNK ⤒㊰ࡀTNF 㑇ఏᏊ㌿
㸪ࡲࡓ p38 ⤒㊰ࡀ㌿ᚋࡢㄪ⠇㐣⛬㛵ࡋ࡚࠸ࡿ⪃࠼ࡽࢀࡓ (56)ࠋ
Celastrol࡛ฎ⌮ࡋࡓ⣽⬊ࡘ࠸࡚ࡣ㸪TNF-α ࡢ⏘⏕ࡣࡃㄆࡵࡽࢀ࡞ࡗࡓࡇ
ࡽ㸪NOྠᵝNF-κBࡢάᛶࡀTNF-αࡢ⏘⏕యⓗ㛵ࢃࡗ࡚࠸ࡿࡇ
ࡀ♧၀ࡉࢀࡓࠋ࡞࠾㸪ࡇࢀࡽࡢ TNF-α ⏘⏕ࡘ࠸࡚㸪SB203580 ཬࡧ SP600125
࡛ฎ⌮ࡋࡓ⣽⬊ࡘ࠸࡚ࡣHTฎ⌮ࡼࡗ࡚పୗഴྥࢆ♧ࡋ㸪U0126࡛ฎ⌮ࡋࡓ
⣽⬊ࡘ࠸࡚ࡣ㸪HTࡼࡗ࡚⃰ᗘ౫Ꮡⓗ᭷ពᢚไࡉࢀࡓࠋ➨1❶࠾࠸࡚㸪
32
HTࡣTNF-α ⏘⏕ᑐࡋᢚไⓗ࡞స⏝ࢆ♧ࡋࡓࡇࡽ㸪ᅇࡢ⤖ᯝࡣ๓ᅇࡢ⤖
ᯝ୍⮴ࡍࡿࡶࡢ࡛࠶ࡗࡓࠋTNF-α⏘⏕ᢚไ㛵ࡋ࡚㸪HT ࡣU0126㸪SB203580 ཬࡧSP600125┦ຠᯝࢆ♧ࡋࡓᤊ࠼ࡿ㸪HTࡢస⏝ⅬࡣERK1/2㸪p38㸪JNK
⤒㊰ୖ௨እ࠶ࡿ⪃࠼ࡽࢀࡓࠋ࡞࠾㸪Western blotting࠾࠸࡚㸪U0126ฎ⌮᮲
௳ୗ࡛ HT ฎ⌮ࡼࡿ pERK1/2 ࡢᢚไࡣㄆࡵࡽࢀ࡚࠾ࡽࡎ㸪HT ࡢస⏝Ⅼࡀ
ERK1/2⤒㊰ୖ↓࠸࠸࠺௬ㄝࢆࡅࡿ⤖ᯝ࡛࠶ࡗࡓࠋ
PGE2㛵ࡋ࡚㸪U0126 (ERK1/2⤒㊰㜼ᐖ)㸪SB203580 (p38㜼ᐖ) ཬࡧSP600125 (JNK㜼ᐖ) ࡛ฎ⌮ࡋࡓ⣽⬊࡛ࡣ㸪LPSㄏⓎᛶࡢPGE2⏘⏕ࡣinhibitorฎ⌮ࡋ࡞
ࡗࡓ⣽⬊ẚ㍑ࡋ࡚1/5⛬ᗘᢚไࡉࢀࡓࠋTNF-αྠᵝPGE2ࡀMAPKࡼࡾ
ไᚚࡉࢀ࡚࠸ࡿࡇࡣከᩘሗ࿌ࡉࢀ࡚࠸ࡿ (57–59)ࠋࡲࡓ㸪NO TNF-α ྠᵝ
celastrol࡛ฎ⌮ࡋࡓ⣽⬊ࡘ࠸࡚ࡣ㸪LPSㄏⓎᛶࡢPGE2⏘⏕ቑຍࡣㄆࡵࡽࢀ࡞
ࡗࡓࠋࡇࢀࡽࡢ⤖ᯝࡽ㸪LPSㄏⓎᛶࡢ⅖ࡣNF-κB⤒㊰ࡀయ࡛㛵ࢃࡗ࡚࠸
ࡿࡇࡀ☜ㄆࡉࢀࡓࠋ࡞࠾㸪TNF-α ␗࡞ࡾ㸪ࡇࢀࡽࡢ PGE2⏘⏕ࡣ㸪U0126㸪
SB203580ཬࡧ SP600125ฎ⌮⣽⬊ࡢ࠸ࡎࢀ࠾࠸࡚ࡶ㸪HT ࡼࡗ࡚⃰ᗘ౫Ꮡⓗ
ᢚไࡉࢀࡓࠋ
➨2❶࠾ࡅࡿ⤖ᯝࡼࡾ㸪HTࡢస⏝Ⅼࡣ ERK1/2㸪p38㸪JNK⤒㊰ୖࡣ↓
ࡃ㸪ࡘ NF-κB ⤒㊰ࡼࡾࡶୗὶ࠶ࡿྍ⬟ᛶࡀ㧗࠸ࡇࡀ᫂ࡽ࡞ࡗࡓ (Fig.
11)ࠋࡲࡓ㸪HTࡣ㸪NO㸪TNF-αཬࡧPGE2⏘⏕ᑐࡋ࡚ᢚไⓗാࡃࡇࡀ☜
ࡵࡽࢀࡓࠋ➨1❶ཬࡧ➨2❶ࡢ⤖ᯝࡽ㸪HTࡣNSAIDsࡸCOX-2㜼ᐖ➼ࡢ᪤
Ꮡࡢᢠ⅖⸆ࡣ␗࡞ࡿ࣓࢝ࢽࢬ࣒࡛ᢠ⅖స⏝ࢆ♧ࡍࡇࡀ᫂ࡽ࡞ࡗࡓࠋ ḟ❶࡛ࡣ㸪ື≀ࣔࢹࣝࢆ⏝࠸࡚HTࡢᢠ⅖స⏝ࢆホ౯ࡋࡓࠋ
33 ᑠᣓ
➨1❶࠾࠸࡚㸪HT ࡀ㸪࣐࢘ࢫ࣐ࢡࣟࣇ࣮ࢪᵝ⣽⬊ᰴRAW264.7 ࠾࠸࡚
LPS่⃭࡛ㄏᑟࡉࢀࡿNOཬࡧPGE2⏘⏕ࢆᢚไࡍࡿࡇࢆ᫂ࡽࡋࡓࠋ HTࡀస⏝ࡍࡿࢩࢢࢼࣝఏ㐩⤒㊰ࢆ᫂ࡽࡍࡿࡓࡵ㸪NF-κBཬࡧERK1/2㸪p38㸪 JNKࡢ3ࡘMAPKᑐࡋ࡚㸪inhibitorࢆ⏝࠸࡚ྛ⤒㊰ࢆ㜼ᐖࡋࡓ㝿ࡢHTࡢస⏝
ࡢኚࢆホ౯ࡋࡓࠋ
⤖ᯝ㸪MAPKࡢ㜼ᐖࡼࡾ TNF-α ཬࡧPGE2ࡢ⏘⏕ࡣపୗࡋ㸪HTࡢฎ⌮ࡼ
ࡾ⏘⏕㔞ࡣࡉࡽపୗࡋࡓࠋࡲࡓ㸪NOࡢ⏘⏕㔞ࡣMAPKࡢ㜼ᐖࡼࡾኚࡋ࡞
ࡗࡓࡀ㸪HTࡢฎ⌮ࡼࡾపୗࡋࡓࠋᮏ⤖ᯝࡼࡾ㸪HTࡢస⏝ⅬࡣERK1/2㸪p38㸪 JNK⤒㊰ୖࡣ↓ࡃ㸪ࡘNF-κB⤒㊰ࡼࡾࡶୗὶ࠶ࡿྍ⬟ᛶࡀ㧗࠸ࡇࡀ᫂ࡽ
࡞ࡗࡓࠋ➨1❶ࡢ⤖ᯝࢆ㋃ࡲ࠼ࡿ㸪HTࡣ12.5 μg/mLࡢ⃰ᗘ࡛ࡣNF-κBࢩ ࢢࢼࣝఏ㐩⤒㊰ࢆᢚไࡍࡿࡇ࡛㸪iNOS㸪COX-2ཬࡧIL-1β㑇ఏᏊࡢⓎ⌧ࢆᢚไ
ࡋ㸪12.5 μg/mLࡼࡾప࠸⃰ᗘ࡛ࡣNF-κB⤒㊰ࡼࡾࡶୗὶ࠶ࡿࢩࢢࢼࣝ⤒㊰స
⏝ࡋ࡚࠸ࡿ⪃࠼ࡽࢀࡓࠋ
ࡲࡓ㸪HTࡣ㸪NOཬࡧPGE2⏘⏕ᑐࡋ࡚ࡣᢚไⓗാࡃࡀ㸪TNF-αᑐࡋ࡚
ࡣస⏝ࢆ♧ࡉ࡞࠸Ⅼࡀ☜ࡵࡽࢀࡓࠋࡇࢀࡣ᪤Ꮡࡢᢠ⅖⸆࡛࠶ࡿ NSAIDs ࡸ
COX-2㜼ᐖࡣぢࡽࢀ࡞࠸స⏝࡛࠶ࡗࡓࠋ
➨1 ❶ཬࡧ➨2 ❶ࡢ⤖ᯝࡽ㸪HT ࡣ᪤Ꮡࡢᢠ⅖⸆ࡣ␗࡞ࡿ࣓࢝ࢽࢬ࣒࡛
ᢠ⅖స⏝ࢆ♧ࡍࡇࡀ᫂ࡽ࡞ࡗࡓࠋḟ㸪ື≀ࣔࢹࣝࢆ⏝࠸࡚HTࡢᢠ⅖
స⏝ࢆホ౯ࡋࡓࠋ
34 ᅗ⾲
Fig. 7. NO production by RAW264.7 cells cultured with LPS (0.25 μg/mL) and HT (0–12.5 μg/mL) for 24 h following the treatment with cellular signaling pathway inhibitors.
Cells were treated with (A) 10 μM SB203580, (B) 10 μM SP600125, (C) 10 μM U0126, or (D) 10 μM celastrol for 2 h before the LPS stimulation. NO concentration in the culture supernatant of RAW264.7 cells was measured by Griess method. Mean ± S.E. *P
< 0.05, **P < 0.01, ***P < 0.001 (Williams’ multiple comparison test) (n = 3).
SB203580 U0126
SP600125 Celastrol
35
Fig. 8. TNF-α production by RAW264.7 cells cultured with LPS (0.25 μg/mL) and HT (0–12.5 μg/mL) for 24 h following the treatment with cellular signaling pathway inhibitors.
Cells were treated with (A) 10 μM SB203580, (B) 10 μM SP600125, (C) 10 μM U0126, or (D) 10 μM celastrol for 2 h before the LPS stimulation. TNF-α concentration in the culture supernatant of RAW264.7 cells was measured by ELISA. Mean ± S.E. *P < 0.05,
***P < 0.001 (Williams’ multiple comparison test) (n = 3).
SB203580
SP600125
U0126
Celastrol
36
Fig. 9. PGE2 production by RAW264.7 cells cultured with LPS (0.25 μg/mL) and HT (0–12.5 μg/mL) for 24 h following the treatment with cellular signaling pathway inhibitors.
Cells were treated with (A) 10 μM SB203580, (B) 10 μM SP600125, (C) 10 μM U0126, or (D) 10 μM celastrol for 2 h before the LPS stimulation. PGE2 concentration in the culture supernatant of RAW264.7 cells was measured by ELISA. Mean ± S.E. *P < 0.05,
**P < 0.01, ***P < 0.001 (Williams’ multiple comparison test) (n = 3).
SB203580
SP600125
U0126
Celastrol
37
Fig. 10. Expression of pNF-κB (A), pERK1/2 (B) in RAW264.7 cells stimulated with LPS (0.25 μg/mL) and HT (0–12.5 μg/mL) following the treatment with cellular signaling pathway inhibitors.
Western blotting analysis was used to determine the levels of these proteins in the cells treated for 15 min. Representative results of duplicate experiments are shown. Cells were treated with (A) 10 μM celastrol or (B) 10 μM U0126 for 2 h before the LPS stimulation.
The graph shows the relative protein expression levels measured by Quantity One Ver 4.5.
Celastrol
U0126
38
Fig. 11. Possible mechanism of anti-inflammatory activity of HT
39
➨3❶
ࣛࢵࢺ࢝ࣛࢤࢽࣥ⫥ᾋ⭘ࣔࢹࣝࢆ⏝࠸ࡓhydroxytyrosolࡢᢠ⅖స⏝ࡢホ౯
40
⥴ゝ
⌧ᅾ㸪㛵⠇࣐ࣜ࢘ࢳཬࡧኚᙧᛶ㛵⠇࡞៏ᛶ⅖ᛶ⑊③ᑐࡋ࡚᪤Ꮡࡢ
NSAIDsཬࡧCOX-2㑅ᢥⓗ㜼ᐖྠ➼௨ୖࡢᢠ⅖࣭㙠③ຠᯝࢆ♧ࡋ㸪ᾘ⟶
㞀ᐖ➼ࡢస⏝ࣜࢫࢡࡀ㍍ῶࡉࢀࡓ᪂つᾘ⅖㙠③⸆ࡢ㛤Ⓨࡀᮃࡲࢀ࡚࠸ࡿ (60)ࠋ ᡃࠎࡣin vitro◊✲࠾࠸࡚㸪HTࡀ㸪LPSㄏⓎᛶࡢiNOS㸪COX-2ཬࡧIL-1β㑇 ఏᏊࡢⓎ⌧ࢆᢚไࡋ㸪⤖ᯝࡋ࡚ NO ཬࡧ PGE2ࡢ⏘⏕㔞ࢆῶᑡࡉࡏࡿࡇࢆぢ ฟࡋࡓࠋᮏ◊✲࡛ࡣ㸪HTࡢ᪂つᾘ⅖㙠③⸆ࡋ࡚ࡢྍ⬟ᛶࢆぢᤣ࠼㸪in vivo
࠾ࡅࡿᢠ⅖άᛶࢆホ౯ࡍࡿࡇࢆ┠ⓗࡋࡓࠋ
࢝ࣛࢤࢽࣥㄏⓎࣛࢵࢺ⫥ᾋ⭘ࣔࢹࣝࡣ㸪ᢠ⅖⸆ࡢຠᯝࢆホ౯ࡍࡿࡢ㐺ࡋ࡚
࠸ࡿ⪃࠼ࡽࢀ࡚࠸ࡿ (61, 62)ࠋ࢝ࣛࢤࢽࣥㄏⓎࣛࢵࢺ⫥ᾋ⭘ࣔࢹࣝ࠾ࡅࡿ⭘
⬽ࡣᛴᛶⁱฟࡼࡿ⭘⬽࡛࠶ࡿ (63)ࠋ࢝ࣛࢤࢽࣥࡣ PG㸪ࣟࢥࢺ࢚ࣜࣥ㸪ࣄࢫ ࢱ࣑ࣥ㸪ࢭࣟࢺࢽࣥ㸪ࣈࣛࢪ࢟ࢽࣥཬࡧTNF-αࢆྵࡴ⅖ㄏⓎᛶ࣓ࢹ࢚࣮ࢱ࣮
ࡢᨺฟࢆ่⃭ࡍࡿࠋࡇࢀࡽࡢ⅖ㄏⓎᛶ࣓ࢹ࢚࣮ࢱ࣮ࡼࡾ㸪ᒁᡤ࡛ᚤᑠᚠ⎔
⣔ࡢᣑᙇ⾑㸪⾑⟶㏱㐣ᛶࡢஹ㐍ࡀ㉳ࡇࡾ㸪⥆࠸࡚ⓑ⾑⌫ࡢ⾑⟶እ࡛ࡢ㐟㉮ࡀ
㉳ࡇࡿࠋ⾑⟶㏱㐣ᛶஹ㐍ࡣ⾑₢⺮ⓑࢆకࡗ࡚Ỉࡀ⾑⟶ࡢእ₃ࢀฟࡍࡇࡼࡗ
࡚㉳ࡇࡿࠋࡇࢀࡼࡾᢞᚋ5㛫௨ෆ㐍⾜ᛶࡢᾋ⭘ࢆㄏⓎࡍࡿ (61)ࠋ ᮏ◊✲࡛ࡣ㸪࢝ࣛࢤࢽࣥㄏⓎࣛࢵࢺ⫥ᾋ⭘ࣔࢹࣝࢆ⏝࠸࡚HTࡢᢠ⅖άᛶࢆ
ホ౯ࡋࡓࠋຍ࠼࡚㸪௦⾲ⓗ࡞NSAIDsࡢࣥࢻ࣓ࢱࢩࣥ (64) ཬࡧ㑅ᢥⓗCOX-2 㜼ᐖ࡛࠶ࡿࢭࣞࢥ࢟ࢩࣈ (65) ࡞ࡢ᪤Ꮡ⒪⸆᭷ຠᛶࢆẚ㍑ࡋࡓࠋ
41
ᮦᩱཬࡧ᪉ἲ
ヨ㦂≀㉁ࡢ⏤᮶
HT ࡢ⏤᮶ࡣ➨ 1 ❶ྠᵝ࡛࠶ࡿࠋࣥࢻ࣓ࢱࢩࣥ㸪࢝ࣛࢤࢽࣥཬࡧ፹యࡋ
࡚⏝࠸ࡓ0.5 w/v% ࣓ࢳࣝࢭ࣮ࣝࣟࢫ⁐ᾮ (0.5 % MC) ࡣ㸪ග⣧⸆ᕤᴗᰴᘧ♫
ࡽ㸪ࢭࣞࢥ࢟ࢩࣈࡣᮾிᡂᕤᴗᰴᘧ♫ࡽࡑࢀࡒࢀ㉎ධࡋࡓࠋ
ヨ㦂≀㉁ࡢㄪ〇 (1) HTࡢㄪ〇
100 mg/mLࡢ⁐ᾮࢆ␃Ỉ࡛2ಸᕼ㔘ࡋ࡚㸪50 mg/mLㄪ〇ࡋࡓࠋ
(2) ࣥࢻ࣓ࢱࢩࣥᠱ⃮ᾮࡢㄪ〇
ࣥࢻ࣓ࢱࢩࣥࢆ㐺㔞⛗㔞ࡋ㸪࣓ࣀ࢘ங㖊ࢆ⏝࠸࡚ศ⢊○ࡋ㸪0.5% MCࢆ ᚎࠎຍ࠼࡚3 mg/mLࡢᠱ⃮ᾮ࡞ࡿࡼ࠺⏝ㄪ〇ࡋࡓࠋ
(3) ࢭࣞࢥ࢟ࢩࣈᠱ⃮ᾮࡢㄪ〇
ࢭࣞࢥ࢟ࢩࣈࢆ㐺㔞⛗㔞ࡋ㸪࣓ࣀ࢘ங㖊ࢆ⏝࠸࡚ศ⢊○ࡋ㸪0.5% MC ࢆ ᚎࠎຍ࠼࡚10 mg/mL⁐ᾮࡢᠱ⃮ᾮ࡞ࡿࡼ࠺⏝ㄪ〇ࡋࡓࠋ
(4) ࢝ࣛࢤࢽࣥ⁐ᾮࡢㄪ〇
࢝ࣛࢤࢽࣥࢆ㐺ᙜ㔞⛗㔞ࡋ㸪1.0%࡞ࡿࡼ࠺␃Ỉࢆຍ࠼㸪ࢫࢱ࣮࣮ࣛࢆ⏝
࠸୍࡚ᬌ᧠ᢾࡋ࡚ㄪ〇ࡋࡓࠋ
42 ౪ヨື≀
ᙜヨ㦂ࡣ㸪⛉◊〇⸆ᰴᘧ♫ (⸨ᯞ㸪㟼ᒸ┴㸪᪥ᮏ㸪HS㈈ᅋㄆᐃ␒ྕ㸸15-047) ࡢື≀ᐇ㦂ࡢᐇ㛵ࡍࡿつᐃཬࡧࡑࡢࡢ㛵ಀἲ௧ࢆ㑂Ᏺࡋ㸪ᢎㄆࡉࢀࡓື≀
ᐇ㦂ィ⏬᭩ (チྍ␒ྕ㸸S16-012) ᚑࡗ࡚ᐇࡋࡓࠋ5 㐌㱋ࡢ Sprague–Dawley
(Crl:CD (SD) ) ⣔⤫ࡢ㞝ᛶࣛࢵࢺࢆ᪥ᮏࢳ࣮ࣕࣝࢫ࣭ࣜࣂ࣮ᰴᘧ♫ (ཌᮌ⦾Ṫࢭ
ࣥࢱ࣮㸪⚄ዉᕝ┴㸪᪥ᮏ) ࡽ㉎ධࡋ㸪7 ᪥㛫ᐇ㦂⎔ቃ㥆ࡉࡏࡓࠋ㣫⫱ᐊࡣ
ᗘ23 ± 3 ºC㸪‵ᗘ50 ± 20%タᐃࡋ㸪㣫⫱ᐊෆࡢẼ㢖ᗘࡣ1㛫࠶ࡓࡾ17
ᅇ௨ୖࡋࡓࠋ᫂ᬯࢧࢡࣝࢆ12㛫ࡋ㸪7:00–19:00ࢆ᫂ᮇࡋࡓࠋ1ࢣ࣮ࢪ
(W 450×D 500×H 350mm㸪㔠⥙ࢣ࣮ࢪ) ᙜࡓࡾ1༉ࡢࣛࢵࢺࢆᐜࡋࡓࠋ㥆ᮇ
㛫୰㸪ᇶ♏㣫ᩱ (࢚࢜ࣜࣥࢱࣝ㓝ẕᕤᴗᰴᘧ♫㸪ᮾி㸪᪥ᮏ) ཬࡧୖỈ (⸨ᯞ ᕷ㸪㟼ᒸ┴㸪᪥ᮏ) ࢆ⮬⏤ᦤྲྀࡉࡏࡓࠋ㥆ᚋ㸪ࣛࢵࢺࢆᒙู↓సⅭἲ࡚㸪
᭱᪂ࡢయ㔜ᇶ࡙࠸࡚ྛ⩌ࡾศࡅヨ㦂⏝ࡋࡓࠋ
ᐇ㦂᪉ἲ
⩌ᵓᡂࢆ௨ୗ♧ࡍࠋ
⩌␒ྕ ⩌ ㊊㋲⓶ୗᢞ ヨ㦂≀㉁ ᢞᐜ㔞 ᩘ
1 ↓ฎ⨨ᑐ↷⩌ ⏕⌮㣗ሷᾮ 0.5% MC
10 mL/kg 5 2 㐺ྜᑐ↷⩌
1.0%
࢝ࣛࢤࢽࣥ⁐
ᾮ
0.5% MC
3 HT
500 mg/kg⩌
HT 50 mg/mL 4 ࢭࣞࢥ࢟ࢩࣈ
100 mg/kg⩌
ࢭࣞࢥ࢟ࢩࣈ
10 mg/mL 5 ࣥࢻ࣓ࢱࢩࣥ
30 mg/kg⩌
ࣥࢻ࣓ࢱࢩࣥ
3 mg/mL
43
↓ฎ⨨ᑐ↷⩌ࡣ⾑ᾮᏛ᳨ᰝ㸪⾑ᾮ⏕Ꮫ᳨ᰝ࠾ࡅࡿẚ㍑ᑐ↷ࡋ࡚タᐃࡋࡓࠋ
⏝㔞タᐃ㛵ࡋ࡚㸪HTࡣ2 g/kgࡢ⏝㔞࡛ࣛࢵࢺẘᛶࢆ♧ࡉ࡞࠸ሗ࿌ࡉࢀ࡚
࠸ࡿࡇࡽ (36), ẘᛶᙳ㡪ࡀ↓ࡃ⸆ຠࡀᮇᚅ࡛ࡁࡿ⏝㔞ࡋ࡚500 mg/kgࢆタ ᐃࡋࡓࠋ➨ 4 ❶࠾࠸࡚ࢫࣆࣜࣥࡢే⏝ࡼࡿస⏝ᙳ㡪ࢆ᳨ウࡍࡿࡓࡵ㸪 ࢭࣞࢥ࢟ࢩࣈ㛵ࡋ࡚ࡣ㸪ẚ㍑ᑐ↷ࡋ࡚㸪⸆ຠࢆ♧ࡋࡘ༢⊂ࡢᢞ࡛ࡣᾘ
⟶㞀ᐖࢆ♧ࡉ࡞࠸᪤▱⏝㔞 (66) ࢆタᐃࡋࡓࠋࣥࢻ࣓ࢱࢩࣥ㛵ࡋ࡚ࡣ㸪㝧ᛶ ᑐ↷ࡋ࡚⸆ຠࢆ♧ࡋࡘ༢⊂ࡢᢞ࡛ᾘ⟶㞀ᐖࢆ♧ࡍ᪤▱⏝㔞 (67) ࢆタᐃ ࡋࡓࠋ
1. ⫥ᐜ✚ ᐃཬࡧᾋ⭘⋡ᾋ⭘ᢚไ⋡ࡢ⟬ฟ
ヨ㦂≀㉁ࢆ10 mL/kgࡢᐜ㔞࡛ࣛࢵࢺᙉไ⤒ཱྀᢞࡋࡓࠋ᪤ሗ (68) ᇶ࡙ࡁ ᢞ1㛫ᚋ㸪1.0%࢝ࣛࢤࢽࣥ⁐ᾮ0.1mLࢆࣛࢵࢺྑᚋ㊊ࡢ㊊㋲⓶ୗᢞࡋࡓࠋ
⫥ᐜ✚ࡣ㸪ࣉࣞࢳࢫ࣓࣮ࣔࢱ࣮ (Model 7141, Ugo Basile, Milan, Italy) ࢆ⏝࠸࡚㸪
࢝ࣛࢤࢽࣥᢞ๓ཬࡧᢞᚋ1㸪2㸪3㸪4ཬࡧ5㛫 ᐃࡋࡓࠋ᪤ሗ (61) ᇶ
࡙ࡁ௨ୗࡢᘧࢆ⏝ࡋ࡚㸪ᾋ⭘⋡ (% swelling) ཬࡧᾋ⭘ᢚไ⋡ (% inhibition) ࢆ ィ⟬ࡋࡓࠋ
% swelling = (VX − V0) /V0 × 100 (x = 1–5)
% Inhibition = (Control – Test) /Control × 100
V0ཬࡧVXࡣࡑࢀࡒࢀ࢝ࣛࢤࢽࣥᢞ๓ (0 㛫) ཬࡧ1㸪2㸪3㸪4ཬࡧ5㛫 ᚋࡢᚋ⫥ᐜ✚࡛࠶ࡾ㸪Control ཬࡧ Test ࡣࡑࢀࡒࢀ㐺ྜᑐ↷⩌ཬࡧヨ㦂≀㉁ᢞ
44
⩌࠾ࡅࡿᖹᆒᐜ✚࡛࠶ࡿࠋ
2. ⫥⤌⧊ࡢ⌮⤌⧊Ꮫⓗ᳨ᰝ
ࡍ࡚ࡢື≀ࢆ㸪⫥ᐜ✚ࡢ᭱⤊ ᐃᚋ㸪ࢯࣇࣝࣛࣥ྾ධ㯞㓉ୗ࡛⭡㒊㟼⬦
ཬࡧື⬦ࢆษ᩿ࡋ࡚ᨺ⾑ࡼࡾᏳᴦẅࡋࡓࠋᏳᴦẅᚋ㸪⫥⤌⧊ࢆ᥇ྲྀࡋ10 vol%
୰ᛶ⦆⾪࣐࣍ࣝࣜࣥᾮ࡛ᅛᐃᚋ㸪⬺⬡ཬࡧࢠ㓟㺃ࢡ࢚ࣥ㓟ࢼࢺ࣒ࣜ࢘ἲࡼࡿ⬺
⅊ࢆ⾜ࡗࡓࠋᅛᐃᚋ㸪⬺Ỉ㸪⬺⬡㸪ࣃࣛࣇࣥᾐ㏱ࡋ㸪ࣃࣛࣇࣥໟᇙࡋࡓࠋࣃ
ࣛࣇࣥࣈࣟࢵࢡࡣⷧษࡋ㸪࣐࣊ࢺ࢟ࢩ࣭࢚ࣜࣥ࢜ࢪࣥ (H.E.) ᰁⰍࢆࡋࡓࠋ ᰁⰍᶆᮏࢆ᳨㙾ࡋ㸪௨ୗࡢᇶ‽ᚑ࠸㸪ΰྜ⣽⬊ᾐ₶ཬࡧᾋ⭘ࡢ⛬ᗘࡘ࠸࡚
⌮Ꮫⓗ࡞ࢢ࣮ࣞࢻࡅࢆ⾜ࡗࡓ㸸0, negative; 1, minimal; 2, slight; 3, moderate.
3. ⾑ᾮᏛⓗ᳨ᰝ
Ᏻᴦẅ EDTA-2K ࢆ⏝࠸࡚᥇ྲྀࡋࡓ⾑ᾮࢆ⏝࠸࡚㸪⥲ྜ⾑ᾮ᳨ᰝ⨨
(ADVIA120, Bayer AG, Leverkusen, Germany) ࡼࡾ௨ୗࡢ᳨ᰝࢆ⾜ࡗࡓࠋ
࣭⥲ྜ⾑ᾮ᳨ᰝ⨨ (ADVIA120) ࡼࡿ᳨ᰝ㡯┠㸪༢ཬࡧ ᐃ᪉ἲ
㉥⾑⌫ᩘ RBC ×106/μL 2 ゅᗘ䝺䞊䝄䞊䝣䝻䞊䝃䜲䝖䝯䝖䝸 䞊ἲ
࣊ࣔࢢࣟࣅࣥ⃰ᗘ Hemoglobin g/dL 䝅䜰䞁䝯䝖䝦䝰䜾䝻䝡䞁ኚἲ
࣐࣊ࢺࢡࣜࢵࢺ್ Hematocrit % ィ⟬ᘧ䠖 (ᖹᆒ㉥⾑⌫ᐜ✚×㉥⾑
⌫) 䠋10
ᖹᆒ㉥⾑⌫ᐜ✚ MCV fL 2 ゅᗘ䝺䞊䝄䞊䝣䝻䞊䝃䜲䝖䝯䝖䝸 䞊ἲ
ᖹᆒ㉥⾑⌫࣊ࣔࢢࣟࣅࣥ㔞 MCH pg ィ⟬ᘧ䠖 (䝦䝰䜾䝻䝡䞁㔞×㉥⾑
⌫ᩘ) 䠋10 ᖹᆒ㉥⾑⌫࣊ࣔࢢࣟࣅࣥ⃰
ᗘ MCHC g/dL ィ⟬ᘧ䠖䝦䝰䜾䝻䝡䞁㔞䠋 (㉥⾑
⌫ᩘ×ᖹᆒ㉥⾑⌫ᐜ✚) ×1000
⾑ᑠᯈᩘ Platelet ×103/μL 2 ゅᗘ䝺䞊䝄䞊䝣䝻䞊䝃䜲䝖䝯䝖䝸
45
䞊ἲ
ⓑ⾑⌫ᩘ WBC ×103/μL 2 ゅᗘ䝺䞊䝄䞊䝣䝻䞊䝃䜲䝖䝯䝖䝸 䞊ἲ
⥙≧㉥⾑⌫ᩘཬࡧⓒศ⋡ Reticulocyte ×103/μL RNA ᰁⰍἲ䛻䜘䜛䝺䞊䝄䞊䝣䝻 䞊䝃䜲䝖䝯䝖䝸䞊ἲ
ࣜࣥࣃ⌫ᩘཬࡧⓒศ⋡ Lymphocyte ×103/μL
䝨䝹䜸䜻䝅䝎䞊䝊ᰁⰍ䛻䜘䜛䝣䝻 䞊䝃䜲䝖䝯䝖䝸䞊ἲ
ዲ୰⌫ᩘཬࡧⓒศ⋡ Neutrophil㻌 ×103/μL ዲ㓟⌫ᩘཬࡧⓒศ⋡ Eosinophil ×103/μL ዲሷᇶ⌫ᩘཬࡧⓒศ⋡ Basophil ×103/μL ༢⌫ᩘཬࡧⓒศ⋡ Monocyte ×103/μL
4. ⾑ᾮ⏕Ꮫⓗ᳨ᰝ
Ᏻᴦẅ࣊ࣃࣜࣥࣜࢳ࣒࢘ࢆ⏝࠸࡚᥇ྲྀࡋࡓ⾑ᾮࢆ㸪┤ࡕ෭༷㐲ᚰศ㞳ᶵ
(05PR-22㸪᪥❧ᕤᶵᰴᘧ♫㸪ᮾி㸪᪥ᮏ) ࢆ⏝࠸ࡓ㐲ᚰศ㞳 (4 Υ㸪ẖศ3,000
ᅇ㌿㸪15ศ㛫) ࢆ⾜࠸㸪⾑₢ࢆศ㞳ࡋࡓࠋᚓࡽࢀࡓ⾑₢ࢆ⏝࠸࡚㸪⏕Ꮫ⮬ືศ ᯒ⨨ (LABOSPECT 006) ࡼࡾ௨ୗࡢ᳨ᰝࢆ⾜ࡗࡓࠋ
࣭⮬ືศᯒ⨨ (LABOSPECT 006) ࡼࡿ᳨ᰝ㡯┠㸪༢ཬࡧ ᐃ᪉ἲ
⥲⺮ⓑ್ T.Protein g/dL ࣅ࢘ࣞࢵࢺἲ
ࣝࣈ್࣑ࣥ Albumin g/dL BCG ἲ
ࣝࣈ࣑࣭ࣥࢢࣟࣈࣜࣥẚ A/G ratio − ⥲⺮ⓑཬࡧࣝࣈ್࣑ࣥࡽ⟬ฟ
ࢫ ࣃ ࣛ ࢠ ࣥ 㓟 ࣑ࣀ ࢺ
ࣛࣥࢫࣇ࢙࣮ࣛࢮάᛶ AST U/L JSCCᶆ‽ᑐᛂἲ
ࣛ ࢽ ࣥ ࣑ ࣀ ࢺ ࣛࣥ ࢫ
ࣇ࢙࣮ࣛࢮάᛶ ALT U/L JSCCᶆ‽ᑐᛂἲ
ࣝ ࢝ ࣜ ࣍ ࢫ ࣇ ࢱ࣮ ࢮ
άᛶ ALP U/L JSCCᶆ‽ᑐᛂἲ
⾑⢾್ Glucose mg/dL ࣊࢟ࢯ࢟ࢼ࣮ࢮ࣭G-6-PDH ἲ
46
⤫ィゎᯒ
⤖ᯝࡣ㸪ᖹᆒ ± ᶆ‽ㄗᕪ (S.E.) ࡋ࡚♧ࡋࡓࠋᐃ㔞ⓗࢹ࣮ࢱ (ᾋ⭘⋡㸪⾑ᾮ Ꮫཬࡧ⾑ᾮ⏕Ꮫ᳨ᰝ㡯┠) ࡘ࠸࡚ࡣ㸪Bartlettࡢ᳨ᐃࡼࡾ➼ศᩓᛶࢆ᳨ウࡋ㸪
➼ศᩓࡢሙྜࡣStudent’s t-test ἲࢆ⏝࠸᳨࡚ᐃࢆ⾜ࡗࡓࠋ➼ศᩓㄆࡵࡽࢀ࡞
ࡗࡓሙྜࡘ࠸࡚ࡣWilcoxon rank sum testࡢ᳨ᐃࢆ⾜ࡗࡓࠋᐃᛶⓗࢹ࣮ࢱ (⌮
ホ౯ࢢ࣮ࣞࢻ) ࡘ࠸࡚ࡣ⣼✚χ2᳨ᐃࢆ⾜ࡗࡓࠋ᭷ពᕪ᳨ᐃࡢ᭱ప᭷ពỈ‽ࡣ୧
ഃ5%ࡋࡓࠋᾋ⭘⋡ཬࡧ⌮ホ౯ࢢ࣮ࣞࢻ㛵ࡋ࡚ࡣ㸪㐺ྜᑐ↷⩌ (2⩌) ヨ 㦂≀㉁ᢞ⩌ (3–5 ⩌) ࡢ㛫࡛⤫ィࢆᐇࡋࡓࠋ⾑ᾮᏛཬࡧ⾑ᾮ⏕Ꮫ᳨ᰝ
㛵ࡋ࡚ࡣ㸪↓ฎ⨨ᑐ↷⩌ (1⩌) 㐺ྜᑐ↷⩌ (2⩌) ཬࡧ㐺ྜᑐ↷⩌ (2⩌) ヨ 㦂≀㉁ᢞ⩌ (3–5⩌) ࡢ㛫࡛⤫ィࢆᐇࡋࡓࠋ⤫ィゎᯒࡣSASཬࡧEXSUS
ࢆ⏝ࡋࡓࠋ
ᒀ⣲❅⣲್ Ureanitrogen mg/dL ࣮࢘ࣞࢮ࣭GlDH ἲ㸪ࣥࣔࢽ
ᾘཤἲ
ࢡࣞࢳࢽ್ࣥ Creatinine mg/dL ࢡࣞࢳࢽࢼ࣮ࢮ࣭HMMPSἲ
⥲ࢥࣞࢫࢸ್࣮ࣟࣝ Cholesterol mg/dL ࢥ ࣞ ࢫ ࢸ ࣟ ࣮ ࣝ ࢜ ࢟ ࢩ ࢲ ࣮ ࢮ ࣭
HMMPSἲ
ࢺࣜࢢࣜࢭࣜࢻ್ Triglyceride mg/dL GPO࣭HMMPSἲ㸪ࢢࣜࢭࣜࣥᾘཤ ἲ
ࣜࣥ⬡㉁್ Phospholipid mg/dL ࢥࣜࣥ࢜࢟ࢩࢲ࣮ࢮ࣭DAOS ἲ
↓ᶵ್ࣜࣥ Inor-P mg/dL PNP࣭XDH ἲ ࢼࢺ್࣒ࣜ࢘ Sodium mEq/L ࢜ࣥ㑅ᢥ㟁ᴟἲ
್࣒࢝ࣜ࢘ Potassium mEq/L ࢜ࣥ㑅ᢥ㟁ᴟἲ ࢡ್࣮ࣟࣝ Chloride mEq/L ࢜ࣥ㑅ᢥ㟁ᴟἲ
47
⤖ᯝ
1. ࣛࢵࢺ࢝ࣛࢤࢽࣥ⫥ᾋ⭘ࣔࢹࣝ࠾ࡅࡿHTࡢᢠᾋ⭘స⏝
࢝ࣛࢤࢽࣥㄏⓎࣛࢵࢺ⫥ᾋ⭘ࣔࢹࣝ࠾ࡅࡿHTࡢᢠ⅖స⏝ࢆホ౯ࡋࡓࠋ⫥
ᐜ✚ࡣ㸪ࡍ࡚ࡢ⩌࠾࠸࡚࢝ࣛࢤࢽࣥᢞᚋቑຍࡋࡓ (Fig. 12)ࠋHT ᢞ⩌
࡛ࡣ㸪࢝ࣛࢤࢽࣥᢞ 2㸪3 ཬࡧ 5 㛫ᚋ࠾࠸࡚㸪㐺ྜᑐ↷⩌ࡼࡾࡶ᭷ពᾋ
⭘⋡ࡀపୗࡋࡓ (p <0.05)ࠋࢭࣞࢥ࢟ࢩࣈᢞ⩌࡛ࡣ㸪࢝ࣛࢤࢽࣥᢞ1㸪2ཬࡧ 3㛫ᚋ࡛㐺ྜᑐ↷⩌ࡼࡾࡶ᭷ពᾋ⭘⋡ࡀపୗࡋࡓ (p <0.05)ࠋࣥࢻ࣓ࢱࢩࣥ
ᢞ⩌࡛ࡣ㸪࢝ࣛࢤࢽࣥᢞ2㸪3㸪4ཬࡧ5㛫ᚋ࡛㐺ྜᑐ↷⩌ࡼࡾࡶ᭷ពᾋ
⭘⋡ࡀపୗࡋࡓ (p <0.01) (Fig. 13)ࠋ
࢝ࣛࢤࢽࣥᢞ1㸪2㸪3㸪4ཬࡧ5㛫ᚋࡢᾋ⭘ᢚไ⋡ࡣ㸪HTᢞ⩌࡛ࡣࡑࢀ
ࡒࢀ 0.34%㸪13.3%㸪19.2%㸪20.9%ཬࡧ 22.9%࡛࠶ࡗࡓࠋࢭࣞࢥ࢟ࢩࣈᢞ⩌࡛
ࡣ49.6%㸪33.4%㸪24.0%㸪17.6%ཬࡧ14.2%㸪ࣥࢻ࣓ࢱࢩࣥᢞ⩌࡛ࡣ52.7%㸪
61.5%㸪52.1%㸪51.6%ཬࡧ47.9%࡛࠶ࡗࡓ (Fig. 14)ࠋ
ᡭᢏⓗ࣑ࢫࡼࡾ㸪㐺ྜᑐ↷⩌ࡢ1༉ࡣ࢝ࣛࢤࢽࣥࢆ㐺ษᢞ࡛ࡁ࡞ࡗ ࡓࠋᚑࡗ࡚㸪ࡇࡢື≀ࡣホ౯ࡽ㝖እࡋࡓࠋᮏᥐ⨨ࡼࡾ1Ḟᦆ್࡞ࡗࡓࡀ㸪
⤫ィゎᯒᚲせ࡞ࢹ࣮ࢱᩘࡣᚓࡽࢀ࡚࠾ࡾ㸪ヨ㦂ࡢಙ㢗ᛶᙳ㡪ࢆཬࡰࡉ࡞࠸
ุ᩿ࡋࡓࠋ
2. ⫥⤌⧊࠾ࡅࡿ⌮⤌⧊Ꮫⓗ᳨ᰝ
࢝ࣛࢤࢽࣥᢞ5㛫ᚋ࠾ࡅࡿ⌮⤌⧊Ꮫⓗホ౯࠾࠸࡚㸪㊊⏥㒊㍍ᗘࡢ ΰྜ⣽⬊ᾐ₶ཬࡧ୰⛬ᗘࡢᾋ⭘ࡀࡍ࡚ࡢື≀࡛ㄆࡵࡽࢀࡓࠋࡋࡋ㸪⌮Ꮫⓗ
48
ࢢ࣮ࣞࢻࡅᇶ࡙ࡃΰྜ⣽⬊ᾐ₶ࡢ⛬ᗘࡣ㸪HT⩌ (p <0.05)㸪ࢭࣞࢥ࢟ࢩࣈ⩌
(p <0.01) ཬࡧࣥࢻ࣓ࢱࢩࣥ⩌ (p <0.05) ࠾࠸࡚㐺ྜᑐ↷⩌ࡼࡾ᭷ពప
ࡗࡓࠋࡉࡽ㸪ᾋ⭘ࡢ⛬ᗘࡣࢭࣞࢥ࢟ࢩࣈ⩌ (p <0.01) ཬࡧࣥࢻ࣓ࢱࢩࣥ⩌ (p
<0.01) ࠾࠸࡚㐺ྜᑐ↷⩌ࡼࡾ᭷ពపࡗࡓ (Fig. 15, Fig. 16)ࠋ
3. ⾑ᾮᏛ᳨ᰝཬࡧ⾑ᾮ⏕Ꮫ᳨ᰝ
⾑ᾮᏛ᳨ᰝ࠾࠸࡚㸪↓ฎ⨨ᑐ↷⩌ẚ㍑ࡋ࡚㸪㐺ྜᑐ↷⩌࡛ࡣዲ୰⌫ᩘࡢ᭷
ព࡞ቑຍཬࡧࣜࣥࣃ⌫ࡢ᭷ព࡞ቑຍࡀㄆࡵࡽࢀࡓ (p <0.05)ࠋ㐺ྜᑐ↷⩌ヨ㦂≀
㉁ᢞ⩌ࡢẚ㍑࡛ࡣ㸪᭷ព࡞ᕪࡣㄆࡵࡽࢀ࡞ࡗࡓ (Table 2)ࠋ
⾑ᾮ⏕Ꮫ᳨ᰝ࠾࠸࡚㸪↓ฎ⨨ᑐ↷⩌ẚ㍑ࡋ࡚㸪㐺ྜᑐ↷⩌࡛ࡣ࣒࢝ࣜ࢘
ࡢ᭷ព࡞పୗࡀㄆࡵࡽࢀࡓࠋ㐺ྜᑐ↷⩌ヨ㦂≀㉁ᢞ⩌ࡢẚ㍑࡛ࡣ㸪HT ⩌
࡛↓ᶵࣜࣥࡢ᭷ព࡞ቑຍ㸪ࢭࣞࢥ࢟ࢩࣈ⩌࡛ࣝࣈ࣑ࣥࡢ᭷ព࡞పୗ㸪ࢫࣃࣛ
ࢠࣥ㓟࣑ࣀࢺࣛࣥࢫࣇ࢙࣮ࣛࢮάᛶ㸪ᒀ⣲❅⣲್㸪↓ᶵࣜࣥ㸪࣒࢝ࣜ࢘ࡢ᭷ព
࡞ቑຍࡀㄆࡵࡽࢀࡓࠋࣥࢻ࣓ࢱࢩࣥ⩌࡛ࡣ㸪⥲⺮ⓑཬࡧࣝࣈ࣑ࣥࡢ᭷ព࡞ప
ୗ㸪↓ᶵࣜࣥ㸪࣒࢝ࣜ࢘ࡢ᭷ព࡞ቑຍࡀㄆࡵࡽࢀࡓ (Table 3)ࠋ
49
⪃ᐹ
ᮏヨ㦂࡛ࡣ㸪ࣛࢵࢺ࢝ࣛࢤࢽࣥ⫥ᾋ⭘ࣔࢹࣝࢆ⏝࠸࡚ HT㸪ࢭࣞࢥ࢟ࢩࣈཬࡧ
ࣥࢻ࣓ࢱࢩࣥࡢᢠ⅖ຠᯝࢆ᳨ウࡋࡓࠋ㐺ྜᑐ↷⩌ㄆࡵࡽࢀࡓ⤒ⓗ࡞⫥ᐜ
✚ࡢቑຍᑐࡋ࡚㸪HT㸪ࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡣ⫥ᐜ✚ࡢቑຍࢆ᭷ព
ᢚไࡋࡓࠋHT ࡢ᭷ព࡞ᢚไຠᯝࡣ࢝ࣛࢤࢽࣥᢞᚋ 2 㛫ࡽㄆࡵࡽࢀ㸪ᢞ
ᚋ5㛫࡛᭱ࡢᢚไస⏝ࢆ♧ࡋࡓ (% inhibition㸸22.9%)ࠋ
ᑐ↷ⓗ㸪ࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡣ㸪ࡑࢀࡒࢀ࢝ࣛࢤࢽࣥᢞᚋ 1
㛫 (% inhibition㸸49.6%) ཬࡧ2㛫 (% inhibition㸸61.5%) ࡽ᭷ព⫥ᾋ⭘
ࢆᢚไࡋࡓࠋHTࡣࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡼࡾᢚไస⏝ࡣᙅࡗࡓࡀ㸪 ࡑࡢຠᯝࡣ㛫ࡶቑຍࡍࡿഴྥ࠶ࡗࡓࠋ୍᪉࡛㸪ࢭࣞࢥ࢟ࢩࣈཬࡧࣥ
ࢻ࣓ࢱࢩࣥࡢຠᯝࡣ㛫ࡶῶᑡࡋࡓࠋᚑࡗ࡚㸪HT ࡣ㐜⾜ᛶࡢᢠ⅖స⏝
ࢆᣢࡘྍ⬟ᛶࡀ࠶ࡾ㸪HT ࢭࣞࢥ࢟ࢩࣈࡲࡓࡣࣥࢻ࣓ࢱࢩࣥࡢ⤌ྜࡏࡼ
ࡾ㸪ᢞึᮇࡽᚋᮇࡲ࡛ᣢ⥆ⓗ࡞ᢠ⅖ຠᯝࡀᚓࡽࢀࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ
⌮⤌⧊Ꮫⓗ᳨ᰝ࠾࠸࡚㸪ΰྜ⣽⬊ᾐ₶ࡢ⛬ᗘࡣ㸪HT㸪ࢭࣞࢥ࢟ࢩࣈཬࡧ
ࣥࢻ࣓ࢱࢩࣥᢞ⩌࡛㐺ྜᑐ↷⩌ࡼࡾ᭷ពపࡗࡓࠋᾐ₶⣽⬊ࡣ㸪ࣜࣥࣃ⌫㸪 ዲ୰⌫㸪ዲ㓟⌫ཬࡧ༢᰾⣽⬊ࡽᵓᡂࡉࢀ࡚࠸ࡓࠋ⅖ᛶ⣽⬊ᾐ₶ཬࡧయᾮࡢ₃ ฟࡣ㸪ࣄࢫࢱ࣑ࣥ㸪ࢭࣟࢺࢽࣥཬࡧPG ࡢᨺฟࡼࡗ࡚ᘬࡁ㉳ࡇࡉࢀ㸪⅖㒊
࡛ࡢᾋ⭘ࢆࡶࡓࡽࡍ (69)ࠋᚑࡗ࡚㸪ࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡘ࠸࡚▱
ࡽࢀࡿࡼ࠺㸪HTࡣ㸪PGE2ࡢ⏘⏕ࢆῶᑡࡉࡏࡿࡇࡼࡗ࡚㸪ࣄࢫࢱ࣑ࣥ㸪ࢭ
ࣟࢺࢽࣥཬࡧPG ࡞ࡢ⅖ᛶ࣓ࢹ࢚࣮ࢱ࣮ࡢ⏘⏕ࢆᢚไࡋ㸪ᢠ⅖ຠᯝࢆ♧
ࡍࡶࡢ⪃࠼ࡽࢀࡓ (70, 71)ࠋ࡞࠾㸪HTࡣΰྜ⣽⬊ᾐ₶ࢆᢚไࡋࡓ୍᪉㸪ᾋ⭘ࢆ
50
ᢚไࡋ࡞ࡗࡓ⌮⏤ࡋ࡚㸪ᾋ⭘ࡣ⅖ࡢึᮇẟ⣽⾑⟶ࡢᣑᙇࡼࡾ⏕ࡌࡿ
ᛂ࡛࠶ࡿࡢᑐࡋ㸪ΰྜ⣽⬊ᾐ₶ࡣ⅖ࡢึᮇ௨㝆⅖ᛶ⣽⬊ࡀ㞀ᐖ㒊㐟
㉮ࡋ࡚࠾ࡇࡿᛂ࡛࠶ࡿࡓࡵ (72)㸪㐜ຠᛶࡢHTࡣ⅖ึᮇᛂ࡛࠶ࡿᾋ⭘ࢆᢚ ไ࡛ࡁ࡞ࡗࡓྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ
࢝ࣛࢤࢽࣥࡣ COX-2 ࢆάᛶࡋ㸪COX-2 ࡣࣛ࢟ࢻࣥ㓟ࢆᇶ㉁ࡋ࡚ PGE2
࡞ࢆ⏕ᡂࡍࡿࡇ࡛⅖ᛂࢆಁ㐍ࡍࡿࠋࡲࡓNO ࡣCOX-2㸪࣍ࢫ࣍ࣜࣃ࣮ࢮ
A2 (PLA2) ࢆ S-ࢽࢺࣟࢩࣝࡍࡿࡇ࡛άᛶࡋ㸪PGE2ࡢ⏘⏕ࢆቑᙉࡉࡏ⅖
ᛂࢆஹ㐍ࡉࡏࡿ (63)ࠋ➨1❶࠾࠸࡚㸪ᡃࠎࡣ HTࡀ NOཬࡧPGE2ࡢ⏘⏕ࢆ
ᢚไࡍࡿࡇࢆ᫂ࡽࡋ࡚࠾ࡾ㸪ࡇࢀࡽࡢస⏝ࡀᮏᐇ㦂࠾࠸࡚ᾋ⭘ᙧᡂᢚไ స⏝⧅ࡀࡗࡓ⪃࠼ࡽࢀࡓࠋ➨ 1 ❶ཬࡧ 3 ❶࠾࠸࡚㸪HT ࡢ NO ཬࡧ PGE2
⏘⏕ᢚไస⏝㸪࢝ࣛࢤࢽࣥᾋ⭘ᙧᡂᢚไస⏝ࢆࡑࢀࡒࢀ☜ㄆࡋ㸪HT ࡢᢠ⅖స
⏝ࡢ᭷ຠᛶࢆ☜ㄆࡍࡿࡇࡀ࡛ࡁࡓࠋࡑࡇ࡛㸪ḟ⌧ᅾࡢ⒪⸆࡛ㄆࡵࡽࢀࡿ
స⏝ࡢ୍ࡘ࡛࠶ࡿᾘ⟶㞀ᐖࡢㄏⓎࡢ᭷↓ࡘ࠸࡚HTࢆホ౯ࡋࡓࠋ
51 ᑠᣓ
ᡃࠎࡣin vitro ◊✲࠾࠸࡚㸪HTࡀ㸪LPSㄏⓎᛶࡢ iNOS㸪COX-2ཬࡧ IL-1β 㑇ఏᏊࡢⓎ⌧ࢆᢚไࡋ㸪⤖ᯝࡋ࡚ NO ཬࡧ PGE2ࡢ⏘⏕㔞ࢆῶᑡࡉࡏࡿࡇࢆ
ぢฟࡋࡓࠋᮏ◊✲࡛ࡣ㸪HT ࡢ᪂つᾘ⅖㙠③⸆ࡋ࡚ࡢྍ⬟ᛶࢆぢᤣ࠼㸪in vivo
࠾ࡅࡿᢠ⅖άᛶࢆホ౯ࡍࡿࡇࢆ┠ⓗࡋࡓࠋ
ヨ㦂≀㉁ࡋ࡚㸪0.5% MC (㐺ྜᑐ↷⩌)㸪HT (500 mg/kg)㸪ࢭࣞࢥ࢟ࢩࣈ (100
mg/kg)㸪ࣥࢻ࣓ࢱࢩࣥ (30 mg/kg) ࢆ㸪࢝ࣛࢤࢽࣥᢞ1㛫๓⤒ཱྀᢞࡋࡓࠋ
࢝ࣛࢤࢽࣥᢞᚋ⫥ᐜ✚ࢆ ᐃࡋ㸪⫥ᐜ✚ࡢኚ㔞ࢆᣦᶆᢠ⅖ຠᯝࢆホ౯ ࡋࡓࠋHTࡣࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡼࡾ⫥ᾋ⭘ᢚไస⏝ࡣᙅࡗࡓࡀ㸪 ࡑࡢຠᯝࡣ㛫ࡶቑຍࡋࡓࠋ୍᪉࡛㸪ࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡢ
ຠᯝࡣ㛫ࡶῶᑡࡋࡓࠋᚑࡗ࡚㸪HT ࡣ㐜⾜ᛶࡢᢠ⅖స⏝ࢆᣢࡘྍ⬟ᛶ ࡀ࠶ࡾ㸪HT ࢭࣞࢥ࢟ࢩࣈࡲࡓࡣࣥࢻ࣓ࢱࢩࣥࡢ⤌ྜࡏࡼࡾ㸪ᢞึᮇ
ࡽᚋᮇࡲ࡛ᣢ⥆ⓗ࡞ᢠ⅖ຠᯝࡀᚓࡽࢀࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ
➨1❶ཬࡧ➨3❶࠾࠸࡚㸪NOཬࡧPGE2⏘⏕ᢚไస⏝㸪࢝ࣛࢤࢽࣥᾋ⭘ᙧᡂ ᢚไస⏝ࢆࡑࢀࡒࢀ☜ㄆࡋ㸪HTࡢᢠ⅖స⏝ࢆ☜ㄆࡍࡿࡇࡀ࡛ࡁࡓࠋHTᢠ
⅖స⏝ࡀㄆࡵࡽࢀࡓࡇࡽ㸪ḟ⌧ᅾࡢ⒪⸆ㄆࡵࡽࢀࡿస⏝ࡢ୰࡛᭱
ࡶၥ㢟࡞ࡿᾘ⟶㞀ᐖㄏⓎࡢ᭷↓ࡘ࠸࡚HTࢆホ౯ࡋࡓࠋ
52 ᅗ⾲ Table 2. Hematology in the rat paw edema model Group No.1 2 3 4 5 Group0.5%MC+Saline 0.5%MC+CarrageenanHT+Carrageenan Celecoxib+CarrageenanIndomethacin+Carrageenan N 4 5 5 5 5 RBC 5.968 ± 0.195 6.268 ± 0.408 6.084 ± 0.208 5.858 ± 0.239 5.978 ± 0.220 Hemoglobin 13.60 ± 0.31 14.00 ± 0.78 13.72 ± 0.32 13.18 ± 0.34 13.64 ± 0.42 Hematocrit40.14 ± 0.85 41.58 ± 2.03 41.60 ± 1.31 39.38 ± 0.98 40.10 ± 1.10 MCV 67.26 ± 1.75 66.43 ± 2.64 68.42 ± 2.03 67.26 ± 2.39 67.12 ± 1.62 MCH 22.76 ± 0.42 22.33 ± 1.03 22.58 ± 0.81 22.50 ± 0.85 22.80 ± 0.52 MCHC 33.88 ± 0.59 33.63 ± 0.33 33.00 ± 0.47 33.46 ± 0.21 33.98 ± 0.43 platelet 1097.2 ± 214.1 1364.5 ± 124.1 1341.2 ± 92.0 1328.4 ± 160.8 1273.8 ± 122.3 WBC9.490 ± 1.775 9.015 ± 1.766 6.982 ± 0.802 8.486 ± 1.469 8.524 ± 1.110 Reticulocyte537.48 ± 44.43 524.53 ± 46.99 497.66 ± 45.43 467.70 ± 33.36 460.34 ± 38.79 Neutrophil 1.868 ± 0.664 4.743 ± 0.741 a 3.874 ± 0.711 4.128 ± 0.854 4.880 ± 0.851 Lymphocyte7.060 ± 0.938 3.853 ± 1.267 a 2.752 ± 0.432 4.004 ± 0.784 3.292 ± 0.402 Monocyte0.382 ± 0.168 0.280 ± 0.074 0.248 ± 0.079 0.230 ± 0.060 0.224 ± 0.034 Eosinophil 0.134 ± 0.121 0.100 ± 0.008 0.072 ± 0.026 0.080 ± 0.049 0.110 ± 0.037 Basophil 0.026 ± 0.009 0.013 ± 0.005 a 0.010 ± 0.000 0.016 ± 0.009 0.010 ± 0.000 a : p < 0.05 compared with control (Student’s t-test). Each value represents the mean ± standard error of the mean of data from five animals, except for the control group, in which data from only four animals was used.
53
Table 3. Blood chemistry in the rat paw edema model Group No.1 2 3 4 5 Group0.5%MC+Saline 0.5%MC+CarrageenanHT+Carrageenan Celecoxib+CarrageenanIndomethacin+Carrageenan N 4 5 5 5 5 T.Protein 4.98 ± 0.08 5.15 ± 0.17 5.20 ± 0.10 4.96 ± 0.11 4.38 ± 0.28 㼎 Albumin 3.96 ± 0.11 4.00 ± 0.08 4.08 ± 0.04 3.84 ± 0.05 c 3.42 ± 0.26 c A/G ratio3.908 ± 0.408 3.510 ± 0.373 3.662 ± 0.280 3.454 ± 0.325 3.584 ± 0.421 AST92.2 ± 6.8 99.3 ± 4.3 99.0 ± 3.5 104.4 ± 4.3 b 83.8 ± 7.6 ALT43.2 ± 3.6 40.8 ± 3.5 42.8 ± 5.1 44.6 ± 4.2 38.2 ± 5.3 ALP1330.6 ± 358.2 1448.5 ± 487.9 1333.8 ± 240.2 1129.8 ± 173.2 965.4 ± 190.0 Glucose 183.0 ± 10.9 163.5 ± 20.0 159.8 ± 12.1 161.2 ± 20.8 149.0 ± 12.6 Ureanitrogen9.06 ± 1.45 7.45 ± 1.59 8.74 ± 1.27 12.22 ± 1.46 c 16.26 ± 4.78 c Creatinine 0.182 ± 0.018 0.180 ± 0.018 0.196 ± 0.011 0.178 ± 0.013 0.192 ± 0.013 Cholesterol77.4 ± 11.6 79.3 ± 12.1 81.0 ± 10.0 80.0 ± 7.4 71.4 ± 10.5 Triglyceride 50.2 ± 23.7 30.5 ± 9.1 21.8 ± 8.0 38.0 ± 21.5 44.2 ± 27.0 Phospholipid130.0 ± 16.6 134.5 ± 14.8 124.4 ± 11.1 140.2 ± 11.8 126.2 ± 15.0 Inor-P9.44 ± 0.46 8.88 ± 0.50 9.54 ± 0.30 b 9.84 ± 0.52 b 9.32 ± 0.65 Sodium144.6 ± 0.9 144.8 ± 2.1 144.4 ± 1.3 145.0 ± 1.6 142.8 ± 1.5 Potassium4.32 ± 0.29 3.60 ± 0.12 a 3.84 ± 0.34 4.56 ± 0.61 b 4.50 ± 0.37 b Chloride102.8 ± 1.9 103.0 ± 0.8 102.2 ± 0.8 102.6 ± 1.5 103.0 ± 0.7 a : p < 0.05 compared with group 1 (Student’s t-test). b : p < 0.05 compared with group 2 (Student’s t-test). c : p < 0.05 compared with group 2 (Wilcoxon rank sum test). Each value represents the mean ± standard error of the mean of data from five animals, except for the group 2, in which data from only four animals was used.
54
Fig. 12. Image showing the hind paw of rat 1 h after carrageenan or saline injection and treatment with HT and celecoxib.
(A) 0.5 w/v % methyl cellulose and saline injection, (B) 0.5 w/v % methyl cellulose and carrageenan injection, (C) HT and carrageenan injection, (D) celecoxib and carrageenan injection. Indomethacin data was not shown.
55
Fig. 13. The % swelling volume after carrageenan injection and treatment with HT, celecoxib, and indomethacin on rat paw edema 1, 2, 3, 4, and 5 h after carrageenan injection.
% swelling was calculated by the following formula:
% swelling = (VX − V0) /V0 (x = 1–5)
where V0 and VX are the hind paw volumes before (0 h) and 1, 2, 3, 4, and 5 h after carrageenan injection, respectively.
*p < 0.05, **p < 0.01, ***p < 0.001, compared to control (Student’s t-test). Each bar represents the mean ± standard error of the mean of data from five animals, except for the control group, in which data from only four animals was used.
56
Fig. 14. The % Inhibition by HT, celecoxib, and indomethacin in the rat paw edema model.
% Inhibition was calculated by the following formula:
% Inhibition = (Control – Test) /Control × 100
where Control and Test are the mean volumes in the control and test groups, respectively.
57
Fig. 15. Histological staining of rat hind paw tissue sections 5 h after carrageenan injection.
Representative images of tissue injury in (A) 0.5 w/v % methyl cellulose as control, (B) HT, (C) celecoxib, and (D) indomethacin. A shows evident dermal edema (*), and subcutaneous mononuclear cell infiltration (↑). They were weaker in B, C and D. de:
dermis, sc: subcutis.
58
Fig. 16. Histopathological grading of rat hind paw tissue sections 5 h after carrageenan injection.
*p < 0.05, **p < 0.01 compared with control (cumulative χ2 test). Each bar represents the mean ± standard error of the mean of data from five animals, except for the control group, in which data from only four animals was used.
59
➨4❶
Hydroxytyrosolࡢ⤒ཱྀᢞࡼࡿࢫࣆࣜࣥㄏⓎ⫶₽⒆ࡢᙳ㡪
60
⥴ゝ
ࡇࢀࡲ࡛ࡢ◊✲࠾࠸࡚㸪ᡃࠎࡣHTࡀ᪤ᏑࡢNSAIDsཬࡧCOX-2㑅ᢥⓗ㜼ᐖ
ẚ㍑ࡋ࡚ᢠ⅖ຠᯝࡣᙅ࠸ࡶࡢࡢ㸪ᣢ⥆ⓗ࡞⸆⌮స⏝ࡀ࠶ࡿࡇࢆ᫂ࡽ
ࡋࡓࠋ
NSAIDsཬࡧCOX-2㑅ᢥⓗ㜼ᐖࡣ㛵⠇࣐ࣜ࢘ࢳཬࡧኚᙧᛶ㛵⠇➼ࡢ៏ᛶ⅖
ࡢ⑊③⟶⌮ࡢࡓࡵ㛗ᮇᢞࡉࢀࡿࡀ (73)㸪㧗㱋ᝈ⪅࡛ࡣᚰ➽᱾ሰ࡞⾑ᰦᙧᡂ ᛶࡢᝈࢆㄏⓎࡍࡿྍ⬟ᛶࡀᣦࡉࢀ࡚࠾ࡾ㸪⾑ᛶᚰᝈࣜࢫࢡࡢ㧗࠸
ࡣ⾑ᑠᯈจ㞟ᢚไຠᯝඃࢀࡿప⏝㔞ࢫࣆࣜࣥ (low-dose aspirin; LDA) (74) ࡢే⏝ᢞࡀ᥎ዡࡉࢀ࡚࠸ࡿ (60, 75, 76)ࠋ
ࡋࡋ㸪NSAIDsཬࡧCOX-2 㑅ᢥⓗ㜼ᐖࡣ㸪༢ẚ࡚ࢫࣆࣜࣥే⏝
ࡣ⫶⢓⭷㞀ᐖࢆቑᝏࡉࡏࡿࡓࡵే⏝ὀពࡉࢀ࡚࠸ࡿ (76–78)ࠋࡇࢀࡣ㸪
NSAIDs ཬࡧ㑅ᢥⓗ COX-2 㜼ᐖࡢ⸆⌮స⏝ࡀ῝ࡃ㛵ࢃࡗ࡚࠸ࡿࠋCOX-1 ཬࡧ
COX-2ࡣ㸪PGࢆ⏘⏕ࡍࡿ㓝⣲࡛࠶ࡿ (79)ࠋNSAIDsཬࡧ㑅ᢥⓗCOX-2㜼ᐖ
ࡼࡿCOX㜼ᐖࡣ㸪PGࡢࣞ࣋ࣝࢆపୗࡉࡏ㸪⑊③ཬࡧ⅖ࢆᢚไࡍࡿ (70)ࠋࡋ
ࡋ㸪COX-2ࡣ⫶࠾࠸࡚ࡣ⫶⢓⭷ࢆಖㆤࡍࡿPGࢆ⏕ᡂࡍࡿ (80)ࠋCOX-2ࡣ㸪ࣛ
ࢵࢺཬࡧ࣐࢘ࢫࢆ⏝࠸ࡓᐇ㦂࠾࠸࡚㸪⫶₽⒆ࡢ⒵㔜せ࡞ᙺࢆᯝࡓࡍࡇ
ࡀሗ࿌ࡉࢀ࡚࠸ࡿ (81, 82)ࠋ⫶ࡀ⾑≧ែࡢሙྜࡣ㸪COX-2ࡢⓎ⌧ࡣಁ㐍ࡉࢀ
ࡿࡀ㸪NSAIDsࡲࡓࡣ㑅ᢥⓗCOX-2㜼ᐖࡢᢞࡣ㸪ᮏⓎ⌧ࢆᢚไࡋ㸪⾑≧ែ
ࡀஹ㐍ࡋ⫶㞀ᐖࢆᝏࡉࡏࡿ (83)ࠋࡇࢀࡽࡢࡇࡽ㸪ࢫࣆࣜࣥే⏝ࡋ࡚ࡶ
⫶⢓⭷㞀ᐖࢆ⏕ࡌ࡞࠸ᢠ⅖⸆ࡀᮃࡲࢀ࡚࠸ࡿࠋHTࡣ➨ 1 ❶ཬࡧ➨3 ❶ࡢ⤖ᯝ
ࡽ㸪COX-2㜼ᐖస⏝ࡀᙅ࠸ྍ⬟ᛶࡀ♧၀ࡉࢀ㸪ࢫࣆࣜࣥే⏝ࡢ⫶⢓⭷㞀ᐖ
61
ཬࡰࡍᙳ㡪ࡣ NSAIDs ཬࡧ COX-2 㑅ᢥⓗ㜼ᐖẚ࡚㍍ᗘ࡛࠶ࡿࡇࡀᮇ ᚅࡉࢀࡿࠋ௨ୖࡼࡾ㸪ᮏヨ㦂࡛ࡣ㸪࢝ࣛࢤࢽࣥࣔࢹࣝᐇ㦂ྠࡌ⣔⤫㸪㐌㱋ཬࡧ ᛶࡢࣛࢵࢺࢆ⏝࠸㸪༢ᅇ⤒ཱྀᢞࡋࡓ HT㸪NSAIDs ࡛࠶ࡿࣥࢻ࣓ࢱࢩࣥཬࡧ
COX-2 㑅ᢥⓗ㜼ᐖࢭࣞࢥ࢟ࢩࣈࡢࢫࣆࣜࣥㄏⓎ⫶₽⒆ཬࡰࡍᙳ㡪ࡘ࠸
᳨࡚ウࡋࡓࠋ
62
ᮦᩱཬࡧ᪉ἲ
ヨ㦂≀㉁
HT㸪ࢭࣞࢥ࢟ࢩࣈ㸪ࣥࢻ࣓ࢱࢩࣥཬࡧ፹యࡋ࡚⏝࠸ࡓ0.5 % MCࡢ⏤᮶ࡣ
๓㏙ྠᵝ࡛࠶ࡿࠋࢫࣆࣜࣥࡣ㸪ග⣧⸆ᕤᴗᰴᘧ♫ࡽ㉎ධࡋࡓࠋ
ヨ㦂≀㉁ࡢㄪ〇
HT ⁐ᾮ㸪ࢭࣞࢥ࢟ࢩࣈᠱ⃮ᾮ㸪ࣥࢻ࣓ࢱࢩࣥᠱ⃮ᾮࡢㄪ〇᪉ἲࡣ➨ 1 ཬࡧ
➨3❶ྠᵝࡋࡓࠋࢫࣆࣜࣥࡘ࠸࡚ࡣ㸪ࢫࣆࣜࣥࢆ㐺㔞⛗㔞ࡋ㸪࣓ࣀ࢘
ங㖊ࢆ⏝࠸࡚ศ⢊○ࡋ㸪0.5% MCࢆᚎࠎຍ࠼࡚5 mg/mLࡢᠱ⃮ᾮ࡞ࡿࡼ
࠺⏝ㄪ〇ࡋࡓࠋ
౪ヨື≀
⏝࠸ࡓື≀ࡢ⏤᮶ཬࡧ㣫⫱᪉ἲࡣ➨3❶ྠᵝ࡛࠶ࡿࠋ࡞࠾㸪ᙜヨ㦂ࡣ㸪⛉◊
〇⸆ᰴᘧ♫ࡢື≀ᐇ㦂ࡢᐇ㛵ࡍࡿつᐃཬࡧࡑࡢࡢ㛵ಀἲ௧ࢆ㑂Ᏺࡋ࡚㸪 ᢎㄆࡉࢀࡓື≀ᐇ㦂ィ⏬᭩ (チྍ␒ྕ㸸S17-049) ᚑࡗ࡚ᐇࡋࡓࠋ
63 ᐇ㦂᪉ἲ
⩌ᵓᡂࢆ௨ୗ♧ࡍࠋ
1. ⫶ࡢ₽⒆㠃✚ࡢ ᐃཬࡧ⫶ࡢയᐖ㠃✚ࡢ⟬ฟ
ヨ㦂≀㉁ࢆ10 mL/kgࡢᐜ㔞࡛⤒ཱྀᢞࡋࡓ1 㛫ᚋࢫࣆࣜࣥ5 mg/mL⁐
ᾮࢆ10 mL/kgࡢᐜ㔞࡛⤒ཱྀᢞࡋࡓࠋࢫࣆࣜࣥᠱ⃮ᾮᢞᚋ4 㛫➨3❶
ྠ୍ࡢ᪉ἲ࡛ᨺ⾑⮴Ṛࡉࡏࡓࠋ㏿ࡸ⫶ࢆฟࡋ㸪ᄇ㛛㒊ࢆࢡ࣓࡛ࣞࣥࡵ㸪 ᗃ㛛㒊ࡽ2 w/v% ୰ᛶ࣐࣍ࣝࣜࣥᾮ (10 w/v% ୰ᛶ࣐࣍ࣝࣜࣥᾮࢆࣜࣥ㓟⦆⾪
⏕⌮㣗ሷỈ࡛5 ಸᕼ㔘ࡋ࡚ㄪ〇) ࢆ8 mL ሸࡋ࡚ᗃ㛛㒊ࢆࢡ࣓࡛ࣞࣥ㛢ሰࡋࡓࠋ
⫶ࢆ2 w/v% ୰ᛶ࣐࣍ࣝࣜࣥᾮ୰࡛⣙10 ศ㛫ᅛᐃࡋࡓᚋ㸪ᄇ㛛㒊ࡽᙃ୰ኸ
㒊ἢࡗ࡚ᗃ㛛㒊ࡲ࡛ษ㛤ࡋ㸪⫶⢓⭷⾲㠃ࡢ₽⒆ࡢ᭷↓ࢆ☜ㄆࡋࡓࠋ⫶⢓⭷㞀ᐖ
(₽⒆) ࡀㄆࡵࡽࢀࡓಶయࡘ࠸࡚ࡣ㸪⏬ീゎᯒࢯࣇࢺ (ImageJ, ⡿ᅜᅜ❧⾨⏕◊
✲ᡤ) ࢆ⏝࠸࡚₽⒆㠃✚ (mm2) ࢆ ᐃࡋࡓࠋ ᐃࡋࡓ₽⒆㠃✚ࡽ௨ୗࡢィ⟬ᘧ
ࢆ⏝࠸࡚⫶ࡢയᐖ㠃✚ (% gastric damage area) ࢆ⟬ฟࡋࡓࠋ
% gastric damage area = lengths of hemorrhagic lesions (mm) × 100/total stomach area (mm2).
⩌ ࢫࣆࣜࣥᢞ ヨ㦂≀㉁ ᢞᐜ㔞 ᩘ
ᑐ↷⩌
ࢫࣆࣜࣥ
5 mg/mL
0.5% MC
10 mL/kg 5 HT
500 mg/kg⩌
HT 50 mg/mL ࢭࣞࢥ࢟ࢩࣈ
100 mg/kg⩌
ࢭࣞࢥ࢟ࢩࣈ
10 mg/mL
ࣥࢻ࣓ࢱࢩࣥ
30 mg/kg⩌
ࣥࢻ࣓ࢱࢩࣥ
3 mg/mL
64 2. ⾑ᾮᏛⓗ᳨ᰝཬࡧ⾑ᾮ⏕Ꮫ᳨ᰝ
➨3❶ྠᵝࡢ᪉ἲ࡛᥇⾑ཬࡧ ᐃࢆ⾜ࡗࡓࠋ᳨ᰝ㡯┠ࡶ➨3❶ྠᵝ࡛࠶ࡿࠋ
⤫ィゎᯒ
⤖ᯝࡣ㸪ᖹᆒ ± ᶆ‽ㄗᕪ (S.E.) ࡋ࡚♧ࡋࡓࠋ⫶ࡢയᐖ㠃✚ࡘ࠸࡚㸪Bartlett ࡢ᳨ᐃࡼࡾ➼ศᩓᛶࢆ᳨ウࡋ㸪➼ศᩓࡢሙྜࡣStudent’s t-test ἲࢆ⏝࠸᳨࡚ᐃ
ࢆ⾜ࡗࡓࠋ➼ศᩓㄆࡵࡽࢀ࡞ࡗࡓሙྜࡘ࠸࡚ࡣWilcoxon rank sum testࡢ᳨
ᐃࢆ⾜ࡗࡓࠋ⤫ィゎᯒࡣSASཬࡧEXSUSࢆ⏝ࡋࡓࠋ
65
⤖ᯝ
1. ⫶ࡢ₽⒆㠃✚ࡢ ᐃཬࡧ⫶ࡢയᐖ㠃✚ࡢ⟬ฟ
HT ࢫࣆࣜࣥࢆే⏝ࡋࡓሙྜࡢ⫶⢓⭷㞀ᐖࡢࣜࢫࢡࢆホ౯ࡋࡓࠋᑐ↷⩌
࠾࠸࡚㸪ฟ⾑ᛶࡧࡽࢇ௦⾲ࡉࢀࡿ⫶㞀ᐖࡣࢫࣆࣜࣥ༢⊂࡛ࡣㄆࡵࡽࢀ࡞ࡗ ࡓ (Fig. 17 A)ࠋྠᵝ㸪ࢫࣆࣜࣥHTࡢే⏝ᢞ࠾࠸࡚ࡶ⫶㞀ᐖࡣㄆࡵࡽ
ࢀ࡞ࡗࡓ (Fig. 17B)ࠋᑐ↷ⓗ㸪ࢫࣆࣜࣥཬࡧࢭࣞࢥ࢟ࢩࣈࡲࡓࡣࣥࢻ࣓
ࢱࢩࣥࡢే⏝ᢞࡣ㸪᫂░࡞⫶㞀ᐖࢆᘬࡁ㉳ࡇࡋࡓ (Fig. 17C, Fig. 17D)ࠋ⫶ࡢയ ᐖ㠃✚ࡣ㸪ࢭࣞࢥ࢟ࢩࣈࡲࡓࡣࣥࢻ࣓ࢱࢩࣥᢞ⩌࠾࠸࡚㸪᭷ព㧗࠸್ࢆ
♧ࡋࡓ (Fig. 18)ࠋ
2. ⾑ᾮᏛ᳨ᰝཬࡧ⾑ᾮ⏕Ꮫ᳨ᰝ
⾑ᾮᏛⓗ᳨ᰝ࡛ࡣ㸪ࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥᢞ⩌࠾࠸࡚㸪㉥⾑⌫
ᩘཬࡧⓑ⾑⌫ᩘࡀቑຍഴྥ࡛࠶ࡗࡓࠋࡋࡋ㸪HT ᢞ⩌࡛ࡣኚࡀ࡞ࡗࡓ (Table 4, Fig. 19A, Fig. 19B)ࠋ⥙≧㉥⾑⌫ᩘࡣ㸪ࣥࢻ࣓ࢱࢩࣥ⩌࠾࠸࡚ࡢࡳቑ ຍഴྥ࠶ࡗࡓ (Table 4, Fig. 19C)ࠋዲ୰⌫ᩘࡣ㸪ࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩ
ࣥ⩌࡛᭷ពቑຍࡋࡓࡀ㸪HT⩌࡛ࡣኚࡀ࡞ࡗࡓ (Table 4, Fig. 19D)ࠋ
⾑ᾮ⏕Ꮫ᳨ᰝ࡛ࡣ㸪ᑐ↷⩌ẚ㍑ࡋ࡚᭷ព࡞ኚࡣㄆࡵࡽࢀ࡞ࡗࡓ (Table 5)ࠋ
66
⪃ᐹ
LDAࡣ㸪⾑ᰦࢆண㜵ࡍࡿࡓࡵࡢຠᯝⓗ࡞⒪ἲ࡛࠶ࡾ㸪㏆ᖺ㸪ฎ᪉ᩘࡀቑຍ ࡋ࡚࠸ࡿ (73)ࠋࡋࡋ㸪ࢫࣆࣜࣥࡣప⏝㔞࡛ࡶᾘ⟶⢓⭷㞀ᐖࢆㄏⓎࡍࡿྍ⬟
ᛶࡀ࠶ࡾ (77)㸪NSAIDs LDA ࡢ୧᪉ࡀ⫶㞀ᐖࡢࣜࢫࢡࣇࢡࢱ࣮࡞ࡿࡉ
ࢀࡿ (67)ࠋNSAIDs ㄏⓎᛶ⫶⢓⭷㞀ᐖࡣ㸪㸪㓟ᛶ⎔ቃୗ࡛ࡢ⢓⭷⣽⬊┤᥋㞀 ᐖCOX㜼ᐖస⏝ࡼࡿ⫶⢓⭷PGࡢᢚไࡼࡗ࡚ᘬࡁ㉳ࡇࡉࢀࡿ (67, 81, 84)ࠋ
NSAIDs ࡣ㓟ᛶ㸪ሷᇶᛶཬࡧ୰ᛶࡀᏑᅾࡍࡿࡀ㸪ᢠ⅖స⏝ࡀᙉ࠸ࡢࡣ㓟ᛶ
NSAIDs࡛࠶ࡾ㸪ࣥࢻ࣓ࢱࢩࣥࡶ㓟ᛶNSAIDsヱᙜࡍࡿࠋ㓟ᛶNSAIDs ࡣ㓟
ᛶ⎔ቃ࡛࠶ࡿ⫶ෆ⭍࠾࠸࡚㠀࢜ࣥ≧ែ࡛࠶ࡾ㸪ᐜ᫆⫶ෆቨࡽ྾ࡉࢀࡿࠋ
྾ࡉࢀࡓ㓟ᛶNSAIDsࡣ㸪྾࣭࢚ࢿࣝࢠ࣮௦ㅰࡢ㞀ᐖࡽ⢓⭷ୖ⓶ࡢ◚⥢ࢆ
ᘬࡁ࠾ࡇࡍࠋࡲࡓ㸪⢓ᾮࡢỈᛶࢆῶᑡࡉࡏ⫶㓟ࡸ࣌ࣉࢩࣥࡼࡿᒁᡤⓗ⢓⭷㞀 ᐖࢆቑᝏࡍࡿ (86)ࠋPG ࡣ⢓ᾮ⏘⏕㸪㔜Ⅳ㓟ศἪ㸪ᚤᑠᚠ⎔ࢆಁ㐍ࡋ㸪ᾘ⟶⢓
⭷ࡢᜏᖖᛶࢆ⥔ᣢࡍࡿ㔜せ࡞⏕⌮άᛶ≀㉁࡛࠶ࡿࡀ㸪NSAIDs ࡣෆᅉᛶ PG ࡢᢚ ไࢆࡋ࡚⢓⭷㜵ᚚᶵᵓࡢ◚⥢ࢆࡁࡓࡍ (86, 87)ࠋ
୍᪉㸪㑅ᢥⓗ COX-2 㜼ᐖ࡛࠶ࡿࢭࣞࢥ࢟ࢩࣈ㉳ᅉࡍࡿᾘᛶ₽⒆ࡢྜే
(ฟ⾑㸪✸Ꮝཬࡧ㏻㐣㞀ᐖ) ࡢⓎ⏕⋡ࡣࡈࡃഹ࡛࠶ࡿ (88)ࠋࡉࡽ㸪ูࡢ㑅
ᢥⓗ COX-2 㜼ᐖ࡛࠶ࡿࣟࣇ࢙ࢥ࢟ࢩࣈࡼࡿ⒪ᚋࡢ⫶༑ᣦ⭠₽⒆ࡢⓎ⏕
㢖ᗘࡣ㸪㠀≉␗ⓗ COX 㜼ᐖ࡛࠶ࡿࣈࣉࣟࣇ࢙ࣥࡼࡿ⒪ᚋࡢࡑࢀࡼࡾࡶ
᫂ࡽప࠸ (89)ࠋࡇࢀࡽࡢ⤖ᯝᇶ࡙ࡁ㸪COX-2㜼ᐖࡣ㸪NSAIDsࡣ␗࡞
ࡾ⫶⭠ࡢస⏝ࡢ࡞࠸㙠③/ᾘ⅖స⏝ࢆⓎࡍࡿ⪃࠼ࡽࢀ࡚࠸ࡓࠋࡋࡋ㸪ࢫ
ࣆࣜࣥ㑅ᢥⓗ COX-2 㜼ᐖࡢే⏝ࡣ㸪ฟ⾑ᛶ₽⒆ࡢࣜࢫࢡࢆⴭࡋࡃቑຍࡉ
67
ࡏࡿࡇࡀ᫂ࡽ࡞ࡗ࡚࠾ࡾ (90)㸪COX-2㜼ᐖࡢᢞࡶ㧗㱋ᝈ⪅ࡗ࡚ࡣ
ࡁ࡞ࣜࢫࢡ࡞ࡗ࡚࠸ࡿ (85)ࠋ
LDAࡣ㸪≉㧗㱋⪅࠾ࡅࡿᵝࠎ࡞ᝈࡢண㜵ཬࡧ⒪ࡢࡓࡵฎ᪉ࡉࢀ࡚࠸
ࡿࠋࡲࡓ, ࣥࢻ࣓ࢱࢩࣥཬࡧࢭࣞࢥ࢟ࢩࣈࡣ㸪࣐ࣜ࢘ࢳࡢࡼ࠺࡞៏ᛶ⅖ᝈ ࡢ⒪ࡶ⏝ࡉࢀࡿࠋࡋࡋ㸪ࡇࢀࡽࡢ⸆ࢆ㸪LDAᢞ⸆୰ࡢ㧗㱋ᝈ⪅⏝
ࡍࡿ㸪⫶⭠⟶㞀ᐖࢆᘬࡁ㉳ࡇࡍྍ⬟ᛶࡀ࠶ࡿ (91)ࠋᑐ↷ⓗ㸪ᮏ◊✲⤖ᯝ࠾
࠸࡚㸪HTࡣࢫࣆࣜࣥྠᢞࡋ࡚ࡶ⫶㞀ᐖࢆ⏕ࡌࡉࡏ࡞ࡗࡓࠋHTࡣ㸪㣗 ရࡸᰤ㣴⿵ຓ㣗ရࡋ࡚᪤ᕷ㈍ࡉࢀ࡚࠾ࡾ (37)㸪㧗⏝㔞 (2 g/kg) ࡛ࡶࡆࡗṑ㢮
࡛ẘᛶࡣㄆࡵࡽࢀ࡚࠸࡞࠸ (36)ࠋHT ࡀ⫶㞀ᐖࢆᘬࡁ㉳ࡇࡉ࡞࠸⌮⏤ࡣ౫↛᫂
࡛࠶ࡿࡀ㸪➨3❶ࡢ⤖ᯝࡽ㸪in vivo࠾ࡅࡿCOX-2㜼ᐖస⏝ࡣࢭࣞࢥ࢟ࢩࣈཬ ࡧࣥࢻ࣓ࢱࢩࣥࡼࡾࡶHTࡣᙅ࠸ࡇࡀ᫂ࡽ࡞ࡗ࡚࠾ࡾ㸪ࡇࡢࡇࡀ⫶㞀 ᐖࡢ㍍ῶ⧅ࡀࡗ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ
HT ࡢᢠ⅖άᛶࡢᶵᗎࢆゎ᫂ࡍࡿࡓࡵࡣ᭦࡞ࡿ◊✲ࡀᚲせ࡛࠶ࡿࡀ㸪ᮏ◊
✲⤖ᯝࡼࡾHTࡣᢞᚋ㛗㛫㸪స⏝↓ࡃ⸆ຠࢆⓎࡋ⥆ࡅࡿࡇࡀᮇᚅࡉࢀ
ࡿࡓࡵ㸪᪂ࡋ࠸ᢠ⅖⸆ࡋ࡚㛤Ⓨ࡛ࡁࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ
68 ᑠᣓ
HTࡣ➨1❶ཬࡧ➨3❶ࡢ⤖ᯝࡽ㸪COX-2㜼ᐖస⏝ࡀᙅ࠸ࡇࡀ♧၀ࡉࢀ㸪
ࢫࣆࣜࣥే⏝ࡢ⫶⢓⭷㞀ᐖཬࡰࡍᙳ㡪ࡣ NSAIDs ཬࡧ COX-2 㑅ᢥⓗ㜼ᐖ
ẚ࡚㍍ᗘ࡛࠶ࡿࡇࡀᮇᚅࡉࢀࡓࠋᮏ❶࡛ࡣ㸪ࡇࢀࡽࡢ࣓ࣜࢵࢺࢆ⪃៖ࡋ
࡚㸪➨ 3 ❶ྠࡌ⣔⤫㸪㐌㱋ཬࡧᛶࡢࣛࢵࢺࢆ⏝࠸࡚ HT㸪ࣥࢻ࣓ࢱࢩࣥཬࡧ ࢭࣞࢥ࢟ࢩࣈࡢࢫࣆࣜࣥㄏⓎ⫶₽⒆ཬࡰࡍᙳ㡪ࢆ᳨ウࡋࡓࠋ
ẚ㍑ᑐ↷ࡋ࡚⏝࠸ࡓࣥࢻ࣓ࢱࢩࣥཬࡧࢭࣞࢥ࢟ࢩࣈࡀ᫂░࡞⫶㞀ᐖࢆᘬ ࡁ㉳ࡇࡋࡓࡢᑐࡋ㸪HT ࡣࢫࣆࣜࣥྠᢞࡋ࡚ࡶ⫶㞀ᐖࢆ⏕ࡌࡉࡏ࡞
ࡗࡓࠋண ㏻ࡾࡢ⤖ᯝࡀᚓࡽࢀࡓࡇࡼࡾ㸪HTࡣ㸪in vivo࠾ࡅࡿCOX-2㜼ᐖ స⏝ࡀࣥࢻ࣓ࢱࢩࣥཬࡧࢭࣞࢥ࢟ࢩࣈࡼࡾࡶᙅ࠸ࡇࡀ㸪⫶㞀ᐖࡢ㍍ῶ⧅ࡀ ࡗ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ
69
ᅗ⾲ Table 4. Hematology in the rat model of gastric damage after co-administration of aspirin and anti-inflammatory agents Group No.1 2 3 4 Group0.5%MC+AspirinHT+AspirinCelecoxib+AspirinIndomethacin+Aspirin N 5 5 5 5 RBC 6.890 ± 0.124 6.870 ± 0.220 7.024 ± 0.159 7.026 ± 0.083 Hemoglobin 15.92 ± 0.50 15.68 ± 0.44 16.02 ± 0.53 16.14 ± 0.64 Hematocrit44.06 ± 1.47 43.72 ± 0.95 44.22 ± 1.03 44.08 ± 1.26 MCV 63.94 ± 1.30 63.70 ± 2.81 62.92 ± 0.66 62.70 ± 1.45 MCH 23.08 ± 0.53 22.88 ± 1.16 22.78 ± 0.55 22.96 ± 0.82 MCHC 36.10 ± 0.86 35.90 ± 0.42 36.22 ± 0.68 36.62 ± 0.69 platelet 1530.0 ± 21.7 1420.6 ± 57.5 b 1514.0 ± 222.9 1412.2 ± 219.5 WBC6.510 ± 2.703 5.708 ± 1.457 7.166 ± 1.681 7.522 ± 1.216 Reticulocyte389.86 ± 19.29 383.36 ± 22.93 369.40 ± 27.82 406.00 ± 7.81 Neutrophil 0.638 ± 0.182 1.060 ± 0.434 1.236 ± 0.384 a 1.434 ± 0.534 a Lymphocyte5.584 ± 2.434 4.292 ± 1.043 5.594 ± 1.251 5.702 ± 0.925 Monocyte0.166 ± 0.096 0.252 ± 0.104 0.216 ± 0.068 0.240 ± 0.064 Eosinophil 0.052 ± 0.013 0.036 ± 0.015 0.036 ± 0.019 0.050 ± 0.019 Basophil 0.026 ± 0.025 0.016 ± 0.009 0.024 ± 0.011 0.024 ± 0.005 a : p < 0.05 compared with group 1 (Student’s t-test). b : p < 0.05 compared with group 1 (Wilcoxon rank sum test). Each value represents the mean ± standard error of the mean of data from five animals, except for the group 2, in which data from only four animals was used.
70
Table 5. Blood chemistry in the rat model of gastric damage after aspirin co-administration Group No.1 2 3 4 Group0.5%MC+AspirinHT+AspirinCelecoxib+AspirinIndomethacin+Aspirin N 5 5 5 5 T.Protein 5.30 ± 0.19 5.36 ± 0.19 5.24 ± 0.09 5.10 ± 0.20 Albumin 4.06 ± 0.18 4.04 ± 0.11 3.98 ± 0.08 3.94 ± 0.17 A/G ratio3.312 ± 0.432 3.082 ± 0.276 3.166 ± 0.169 3.412 ± 0.282 AST75.8 ± 6.0 79.2 ± 9.7 77.6 ± 5.6 73.6 ± 4.2 ALT21.4 ± 2.1 23.4 ± 2.5 23.2 ± 2.9 20.6 ± 0.9 ALP878.4 ± 149.8 1024.8 ± 177.2 970.2 ± 268.3 931.0 ± 268.3 Glucose 127.4 ± 23.5 147.2 ± 19.1 160.2 ± 22.4 136.4 ± 23.9 Ureanitrogen11.40 ± 2.98 11.90 ± 1.03 14.44 ± 1.80 12.46 ± 3.36 Creatinine 0.170 ± 0.010 0.180 ± 0.019 0.194 ± 0.013 a 0.184 ± 0.027 Cholesterol58.0 ± 10.7 55.8 ± 11.3 58.6 ± 11.8 58.4 ± 10.5 Triglyceride 30.6 ± 12.1 25.0 ± 3.3 36.6 ± 9.3 37.6 ± 19.0 Phospholipid85.6 ± 13.0 81.2 ± 10.1 87.0 ± 11.1 88.0 ± 12.8 Inor-P8.30 ± 0.49 8.56 ± 0.71 8.76 ± 0.28 8.62 ± 0.24 Sodium142.2 ± 0.8 142.2 ± 1.5 141.6 ± 1.1 141.2 ± 0.8 Potassium4.04 ± 0.23 3.82 ± 0.19 4.08 ± 0.08 3.92 ± 0.22 Chloride108.0 ± 2.3 108.4 ± 2.6 106.4 ± 1.5 105.4 ± 0.5 a a : p < 0.05 compared with group 1 (Student’s t-test). Each value represents the mean ± standard error of the mean of data from five animals, except for the group 2, in which data from only four animals was used.
71
Fig. 17. Photographs of rat stomachs 4 h after co-administration of aspirin (50 mg/kg) with (A) 0.5 w/v % methyl cellulose as control, (B) HT, (C) celecoxib, and (D) indomethacin.
72
Fig. 18. The % gastric damage area of rat stomachs 4 h after co-administration of aspirin (50 mg/kg) with 0.5 w/v% methyl cellulose as control, HT, celecoxib, and indomethacin.
*p < 0.05 compared with control (Wilcoxon rank sum test). Each bar represents the mean
± standard error of the mean of data from five animals.
73
Fig. 19. Blood parameters in the rat model of gastric damage after co-administration of aspirin (50 mg/kg) with 0.5 w/v % methyl cellulose as control, HT, celecoxib, and indomethacin.
The number of (A) RBCs, (B) WBCs, (C) reticulocytes, and (D) neutrophils in blood samples collected from the abdominal aorta. *p < 0.05 compared with control (Student’s t-test). Each bar represents the mean ± standard error of the mean of data from five animals.
74
⥲ᣓ
㏆ᖺ㸪ࡀࢇࡸ⏕ά⩦័࡞ࡢ៏ᛶᝈࡢែᙧᡂ㸪៏ᛶ⅖ࡀ῝ࡃ㛵ࢃࡗ
࡚࠸ࡿࡇࡀὀ┠ࡉࢀ࡚࠸ࡿࠋ៏ᛶ⅖࡛ࡣ㸪㓟ࢫࢺࣞࢫ࡞ࡢ่⃭ࢆཷࡅࡓ
⮚ჾࡢ⣽⬊ࡽᨺฟࡉࢀࡿ⮬ᕫ⏤᮶ࡢࡓࢇࡥࡃ㉁ᡂศࡀ㸪࣐ࢡࣟࣇ࣮ࢪࡼࡾ
ㄆ㆑ࡉࢀ㸪ࢧࢺ࢝ࣥ㸪PGE2㸪NOࢆࡣࡌࡵࡋࡓᵝࠎ࡞⅖ᛶ࣓ࢹ࢚࣮ࢱ
࣮ࡀ⏘⏕ࡉࢀࡿ⤖ᯝ㸪ᜏᖖⓗ࡞⅖ᛂࡀㄏᑟࡉࢀࡿࠋࡇࡢࡼ࠺㸪࣐ࢡࣟࣇ
࣮ࢪ៏ᛶ⅖ࡢ㛵㐃ࡀὀ┠ࡉࢀ࡚࠾ࡾ㸪ࡇࢀࡽࡢண㜵ᙺ❧ࡘ᪂つྜ≀ࡢ ᥈⣴ࡀ㐍ࡵࡽࢀ࡚࠸ࡿࠋ
ኳ↛≀⏤᮶≀㉁࡛࠶ࡿHTࡣ࣮࢜ࣜࣈࡢⴥࡸᐇ㇏ᐩྵࡲࢀࡿపศᏊࣇ࢙ࣀ
࣮ࣝྜ≀࡛࠶ࡾ㸪in vitroᐇ㦂࠾࠸࡚ᢠ⅖㸪ᢠ㓟㸪ᢠ࢘ࣝࢫཬࡧᢠ⳦స
⏝ࢆ♧ࡍࡇࡀ᫂ࡽ࡞ࡗ࡚࠸ࡿࡀ㸪ࡑࡢస⏝࣓࢝ࢽࢬ࣒ࡣ౫↛ࡋ࡚᫂࡞
Ⅼࡀከ࠸ࠋࡑࡇ࡛㸪ᮏ◊✲࡛ࡣHTࡢᢠ⅖ຠᯝ㛵ࡋ࡚㸪ࡑࡢస⏝ᶵᗎࢆ᫂ࡽ
ࡍࡿࡶ㸪ື≀ࣔࢹࣝ࠾ࡅࡿᢠ⅖స⏝ࢆ᫂ࡽࡍࡿࡇࢆ┠ⓗ
ࡋࡓࠋ
➨1❶࡛ࡣHTࡘ࠸࡚㸪࣐࢘ࢫ࣐ࢡࣟࣇ࣮ࢪᵝ⣽⬊ᰴRAW264.7ࢆ⏝࠸࡚㸪 LPSㄏⓎᛶ⅖࠾ࡅࡿHTࡢᢠ⅖స⏝ࢆホ౯ࡋࡓࠋࡑࡢ⤖ᯝ㸪HTࡣ12.5 μg/mL ࡢ⃰ᗘ࡛ࡣNF-κBࢩࢢࢼࣝఏ㐩⤒㊰ࢆᢚไࡍࡿࡇ࡛㸪TNF-α㑇ఏᏊⓎ⌧㸪IL-1β
⏘⏕㸪COX-2-PGE2ཬࡧiNOS-NO⣔ࢆᢚไࡋ㸪12.5 μg/mLࡼࡾప࠸⃰ᗘ࡛ࡣูࡢ ࢩࢢࢼࣝఏ㐩⤒㊰㛵ࡍࡿࡇ࡛⅖ᛂࢆㄪ⠇ࡍࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ୍
᪉㸪HTࡣLPSㄏⓎᛶࡢ pERK1/2Ⓨ⌧ࢆᢚไࡋ࡞ࡗࡓࠋHTࡢᢠ⅖ຠᯝࢆ
⌮ゎࡍࡿࡓࡵࡣࡉࡽ࡞ࡿ◊✲ࡀᚲせ࡛࠶ࡿࡀ㸪ᮏ◊✲࡛ࡣ㸪HTࡀLPS่
75
⃭RAW264.7⣽⬊ᑐࡋ࡚ᢠ⅖ຠᯝࢆ᭷ࡍࡿࡇࡀ☜ㄆࡉࢀࡓࠋ
➨2 ❶࡛ࡣ㸪HT ࡀస⏝ࡍࡿࢩࢢࢼࣝఏ㐩⤒㊰ࢆ᫂ࡽࡍࡿࡓࡵ㸪NF-κB ཬ ࡧERK1/2㸪p38㸪JNK ࡢ3 ࡘMAPK ᑐࡋ࡚㸪ࡑࢀࡒࢀࢆ㜼ᐖࡍࡿ inhibitorࢆ
⏝࠸࡚㸪ྛ⤒㊰ࢆ㜼ᐖࡋࡓ㝿ࡢHTࡢస⏝ࡢኚࢆホ౯ࡋࡓࠋ⤖ᯝ㸪MAPKࡢ㜼
ᐖࡼࡾ TNF-α ཬࡧ PGE2ࡢ⏘⏕ࡣపୗࡋ㸪HT ࡢฎ⌮ࡼࡾ⏘⏕㔞ࡣࡉࡽప
ୗࡋࡓࠋࡲࡓ㸪NOࡢ⏘⏕㔞ࡣ MAPKࡢ㜼ᐖࡼࡾኚࡋ࡞ࡗࡓࡀ㸪HT ࡢฎ
⌮ࡼࡾపୗࡋࡓࠋࡇࢀࡽࡢ⤖ᯝࡼࡾ㸪HTࡢస⏝Ⅼࡣ ERK1/2㸪p38㸪JNK ⤒㊰
ୖࡣ↓ࡃ㸪ࡘNF-κB⤒㊰ࡼࡾࡶୗὶ࠶ࡿྍ⬟ᛶࡀ㧗࠸ࡇࡀ᫂ࡽ࡞ࡗ ࡓࠋ➨1❶ࡢ⤖ᯝࢆ㋃ࡲ࠼ࡿ㸪HTࡣ㧗⃰ᗘ࡛ࡣNF-κBࢩࢢࢼࣝఏ㐩⤒㊰ࢆᢚ ไࡍࡿࡇ࡛㸪⅖࣓ࢹ࢚࣮ࢱ࣮㛵㐃㑇ఏᏊࡢⓎ⌧ࢆᢚไࡋ㸪ప⃰ᗘ࡛ࡣ
NF-κB⤒㊰ࡼࡾࡶୗὶ࠶ࡿࢩࢢࢼࣝ⤒㊰స⏝ࡋ㸪⅖Ⓨ⌧ࢆᢚไࡋ࡚࠸ࡿ
⪃࠼ࡽࢀࡓࠋࡲࡓ㸪HTࡣNOཬࡧPGE2⏘⏕ᑐࡋ࡚ࡣᢚไⓗാࡃࡀ㸪TNF-α
ᑐࡋ࡚ࡣᢚไస⏝ࢆ♧ࡉࡎ㸪᪤Ꮡࡢᢠ⅖⸆࡛࠶ࡿNSAIDsࡸCOX-2㜼ᐖ
␗࡞ࡿస⏝࣓࢝ࢽࢬ࣒ࢆᣢࡘྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ
➨ 3 ❶࡛ࡣ㸪࢝ࣛࢤࢽࣥㄏⓎ⫥ᾋ⭘ࣔࢹࣝࢆ⏝࠸࡚㸪HT ࡢᢠ⅖స⏝ཬࡧ
ែࡢ㛵ࢆ᳨ウࡋࡓࠋ࢝ࣛࢤࢽࣥᢞ๓ཬࡧᢞᚋ 1㸪2㸪3㸪4 ཬࡧ 5 㛫
⫥ᐜ✚ࢆ ᐃࡋ㸪⫥ᐜ✚ࡢኚ㔞ࢆᣦᶆᢠ⅖ຠᯝࢆホ౯ࡋࡓ⤖ᯝ㸪HT ࡣࢭ
ࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡼࡾ⫥ᾋ⭘ᢚไస⏝ࡣᙅࡗࡓࡀ㸪ࡑࡢຠᯝࡣ
㛫ࡶቑຍࡋࡓࠋ୍᪉࡛㸪ࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡢຠᯝࡣ㛫
ࡶῶᑡࡋࡓࠋᚑࡗ࡚㸪HTࡣ㐜⾜ᛶࡢᢠ⅖స⏝ࢆᣢࡘྍ⬟ᛶࡀ࠶ࡾ㸪HT ࢭࣞࢥ࢟ࢩࣈࡲࡓࡣࣥࢻ࣓ࢱࢩࣥࡢ⤌ྜࡏࡼࡾ㸪ᢞึᮇࡽᚋᮇࡲ࡛ᣢ
⥆ⓗ࡞ᢠ⅖ຠᯝࡀᚓࡽࢀࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ
76
ຍ࠼࡚➨ 4 ❶࡛ࡣ㸪HT ࡢ⤒ཱྀᢞࡼࡿࢫࣆࣜࣥㄏⓎ⫶₽⒆ࡢᙳ㡪ࢆ᳨
ウࡋࡓࠋẚ㍑ᑐ↷ࡋ࡚⏝࠸ࡓࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡀ᫂░࡞⫶㞀ᐖ
ࢆᘬࡁ㉳ࡇࡋࡓࡢᑐࡋ㸪HT ࡣࢫࣆࣜࣥྠᢞࡋ࡚ࡶ⫶㞀ᐖࢆ⏕ࡌࡉࡏ
࡞ࡗࡓࠋHTࡣ㸪㣗ရࡸᰤ㣴⿵ຓ㣗ရ࡛᪤ᕷ㈍ࡉࢀ࡚࠾ࡾ㸪㧗⏝㔞 (2 g/kg) ࡛
ࡶẘᛶᙳ㡪ࡣㄆࡵࡽࢀ࡚࠸࡞࠸ࠋ➨1❶ཬࡧ➨3❶ࡢ⤖ᯝࡽ㸪in vivo࠾ࡅࡿ
HTࡢCOX-2㜼ᐖస⏝ࡣࢭࣞࢥ࢟ࢩࣈཬࡧࣥࢻ࣓ࢱࢩࣥࡼࡾࡶᙅ࠸ࡇࡀ᫂ࡽ
࡞ࡗ࡚࠾ࡾ㸪ࡇࡢຠᯝࡢᙅࡉࡀ⫶㞀ᐖࡢ㍍ῶ⧅ࡀࡗ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽ
ࢀࡓࠋ
ᢠ⅖άᛶࡢᶵᗎࢆゎ᫂ࡍࡿࡓࡵࡣ᭦࡞ࡿ◊✲ࡀᚲせ࡛࠶ࡿࡀ㸪HT ࡣᢞ
ᚋ㛗㛫㸪స⏝↓ࡃ⸆ຠࢆⓎࡋ⥆ࡅࡿࡇࡀ࡛ࡁࡿࡓࡵ㸪᪂ࡋ࠸ᢠ⅖⸆
ࡋ࡚㛤Ⓨ࡛ࡁࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡓࠋ
77 ㅰ㎡
ᮏ◊✲ࢆ㐙⾜ࡍࡿ࠶ࡓࡾ㸪⤊ጞ᠓ษ࡞ࡿࡈᣦᑟ㸪ࡈ㠴᧡ࢆ㈷ࡾࡲࡋࡓᖏᗈ␆
⏘Ꮫ ⋇་Ꮫ◊✲㒊㛛 ᇶ♏⋇་Ꮫศ㔝 ᑠᕝᬕᏊᩍᤵ῝ࡃឤㅰ࠸ࡓࡋࡲࡍࠋ
ࡲࡓ㸪ᮏ◊✲࠶ࡓࡾ᭷┈࡞ࡿࡈᣦᑟ㸪ࡈຓゝࢆ㡬ࡁࡲࡋࡓᖏᗈ␆⏘Ꮫ ⋇་
Ꮫ◊✲㒊㛛 ᇶ♏⋇་Ꮫศ㔝 㑥ಇᩍᤵ㸪ࢢ࣮ࣟࣂࣝࢢ࣓ࣟࢹࢩࣥ◊✲ࢭ
ࣥࢱ࣮ࡢṊ⏣ὒᖹ≉௵ຓᩍ῝ㅰ࠸ࡓࡋࡲࡍࠋ
ᮏㄽᩥࡢసᡂ࠶ࡓࡾ㸪㈗㔜࡞ࡈຓゝࡈᰯ㜀ࢆ㈷ࡾࡲࡋࡓᖏᗈ␆⏘Ꮫ ⋇
་Ꮫ◊✲㒊㛛 ▼᫂ᩍᤵ㸪ᒾᡭᏛ ㎰Ꮫ㒊 ඹྠ⋇་Ꮫ⛉ Ᏹሯ㞝ḟᩍᤵ㸪ᮾ
ி㎰ᕤᏛ ㎰Ꮫ◊✲㝔 ື≀⏕⛉Ꮫ㒊㛛 బࠎᮌ୍ᩍᤵ㸪ᒱ㜧Ꮫᛂ⏝⏕
≀⛉Ꮫ㒊 ⋇་⸆⌮Ꮫ◊✲ᐊ ᾏ㔝ᖺᘯᩍᤵཌࡃᚚ♩⏦ࡋୖࡆࡲࡍࠋ
ࡲࡓ㸪ᮏ◊✲ࡢᶵࢆ࠼࡚ࡃࡔࡉ࠸ࡲࡋࡓ⛉◊〇⸆ᰴᘧ♫♫㛗 ဴኵ Ặ㸪◊✲㛤Ⓨᮏ㒊㒊㛗 ᐙ⏣ຊẶ㸪◊✲⏬㒊㒊㛗 ⥥㈏Ặ㸪᪂⸆⏕ࢭࣥࢱ࣮
ࢭࣥࢱ࣮㛗 ᑠᾈᬸẶ㸪⸆≀ືែ࣭Ᏻᛶ㒊㒊㛗 ⸃ᕝṇᗈẶ୪ࡧ๓㒊㛗 ⟪
㍯㈼Ặ῝ࡃឤㅰ࠸ࡓࡋࡲࡍࠋ
ᐇ㦂㝿ࡋࡈ༠ຊࡃࡔࡉ࠸ࡲࡋࡓ⛉◊〇⸆ᰴᘧ♫᪂⸆⏕ࢭࣥࢱ࣮ ⸆≀ື
ែ࣭Ᏻᛶ㒊ࡢⓙᵝᚰࡼࡾᚚ♩⏦ࡋୖࡆࡲࡍࠋ
᭱ᚋ㸪⤊ጞከ࡞༠ຊࡢࡶ⚾ࢆᨭ࠼࡚ࡃࢀࡲࡋࡓᐙ᪘ࡣᚰࡽឤㅰࡢព
ࢆ⾲ࡋࡲࡍࠋ
78 ᘬ⏝ᩥ⊩
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