Inhibition of experimental abdominal aortic aneurysm in the rat by use of decoy oligodeoxynucleotides suppressing activity of nuclear factor κB and ets transcription factors

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Inhibition of Experimental Abdominal Aortic

Aneurysm in the Rat by Use of Decoy

Oligodeoxynucleotides Suppressing Activity of

Nuclear Factor

䃙

B and ets Transcription Factors

㸦㌿෗ㄪ⠇ᅉᏊ㹌㹄

䃙

㹀࡜ HWV ࡢ࠾࡜ࡾᆺ᰾㓟་⸆࡟ࡼ

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 ┿ᛶ኱ື⬦⒗ࡢⓎ⏕ࡸ㐍ᒎ࡟ࡣࠊ኱ື⬦ቨࡢࣜࣔࢹࣜࣥࢢ࡟࠾ࡅࡿᙎᛶ⥺⥔ࡢ୺せᡂศ࡛࠶ࡿ࢚ࣛ ࢫࢳࣥࡢ◚ቯࡀ㛵୚ࡍࡿࠋࡇࡢ኱ື⬦ቨ࡟࠾ࡅࡿ࢚ࣛࢫࢳࣥ௦ㅰࡣࠊ࣐ࢺࣜࢵࢡࢫศゎ㓝⣲MMPsࠊ ࣐ࢺࣜࢵࢡࢫศゎ㓝⣲㜼ᐖᅉᏊTIMPsࠊྛ✀ࢧ࢖ࢺ࢝࢖ࣥ࡞࡝ࡀ㛵୚ࡍࡿ」㞧࡞⤌⧊ෆ཯ᛂ࡛࠶ࡿ ࡀࠊMMP-2ࠊMMP-9 ࡞࡝ࡢ MMPs ࡀ୰ᚰⓗᙺ๭ࢆ࡞ࡍ࡜⌮ゎࡉࢀ࡚࠸ࡿࠋ୍᪉ࠊ㌿෗ㄪ⠇ᅉᏊ࡛ ࠶ࡿ NF䃙B ࡸ ets ࡣࠊ኱ື⬦ቨ࡟Ⓨ⌧ࡍࡿ MMPs ࡸ᥋╔ᅉᏊ࡞࡝ࢆࠊ⏘⏕⣽⬊ࢆ௓ࡋ࡚ㄪ⠇ࡋ࡚࠸ ࡿࠋᚑࡗ࡚ࠊࡇࢀࡽࡢ㌿෗ㄪ⠇ᅉᏊࢆ㑇ఏᏊⓗ࡟ไᚚࡍࡿࡇ࡜࡟ࡼࡾࠊMMPs ࡢⓎ⌧ࡀᢚไࡉࢀࡿࡶ ࡢ࡜⪃࠼ࡽࢀࡿࠋ  ᮏ◊✲࡛ࡣࠊࣛࢵࢺ኱ື⬦⒗ࣔࢹࣝ࡟࠾࠸࡚ࠊMMPs ࡢ㌿෗ㄪ⠇ᅉᏊ࡛࠶ࡿ NF䃙B ࡜ ets ࢆྠ᫬ ࡟ᢚไࡍࡿࡓࡵࠊ࠾࡜ࡾᆺ᰾㓟་⸆㸦ࢹࢥ࢖㸧ࢆ⏝࠸࡚኱ື⬦⒗ࡢᢚไຠᯝࢆ᳨ウࡋࡓࠋ  ᪉ ἲ  ࣄࢺ⭡㒊኱ື⬦⒗ቨࡢ᦬ฟᶆᮏ࡛ࡣࠊᢠ NF䃙B p65 ᢠయ࡛ච␿ᰁⰍࡋ࡚ࠊNF䃙B ࡢⓎ⌧ࢆ⑓⌮⤌ ⧊Ꮫⓗ࡟ホ౯ࡋࡓࠋ  ࣛࢵࢺ኱ື⬦⒗ࣔࢹ࡛ࣝࡣࠊୗ⫥ື⬦ࡼࡾ࢝ࢸ࣮ࢸࣝࢆ⭡㒊኱ື⬦ෆ࡟␃⨨ࡋ࡚ࠊࣈࢱ⮅࢚ࣛࢫࢱ ࣮ࢮࢆ㑏ὶࡋ࡚኱ື⬦⒗ࢆసᡂࡋࡓࠋࢹࢥ࢖ࡣࠊᑟධ⏝ࢩ࣮ࢺࢆ⭡㒊኱ື⬦࡟ᕳࡁ௜ࡅ࡚ᑟධࡋࠊFITC ࣛ࣋ࣝࢹࢥ࢖࡟ࡼࡾᑟධ㒊఩ࢆ☜ㄆࡋࡓࠋࢹࢥ࢖ࡣࠊNF䃙B ࡜ ets-1 ྠ᫬ᢚไࡢ࣓࢟ࣛࢹࢥ࢖ࠊࢫࢡ ࣛࣥࣈ࣭࣓ࣝ࢟ࣛࢹࢥ࢖㸦ᑐ↷㸧ࠊNF䃙B ༢⊂ᢚไࡢ NF䃙B ࢹࢥ࢖ࠊࢫࢡࣛࣥࣈ࣭ࣝNF䃙B ࢹࢥ࢖ 㸦ᑐ↷㸧ࡢ㸲⩌࡛ẚ㍑᳨ウࡋࡓࠋ኱ື⬦⒗ࡣࠊᙧែⓗホ౯࡜ࡋ࡚㉸㡢Ἴ᳨ᰝ࡛⒗ᚄኚ໬ࢆ㸲㐌㛫ほᐹ ᚋࠊ᦬ฟࡋࡓࠋࣛࢵࢺ⭡㒊኱ື⬦⒗ࡢ᦬ฟᶆᮏ࡛ࡣࠊศᏊ⏕≀Ꮫⓗホ౯࡜ࡋ࡚ࠊNF䃙B ࡜ ets-1 ࡢ㌿ ෗άᛶࡣ Electrophoretic mobility shift assayEMSAࠊMMPs ⺮ⓑࡢⓎ⌧࡜άᛶࡣ Western blotting,

ᰁⰍࠊMiller's elastin and van Gieson ᰁⰍࢆ⾜࠸ࠊᢠ CD31 ᢠయ࡟ࡼࡿච␿ᰁⰍ࡟ࡼࡾ࣐ࢡࣟࣇ࢓ ࣮ࢪࢆホ౯ࡋࡓࠋ  ⤖ ᯝ  ࣄࢺ⭡㒊኱ື⬦⒗ቨ࡛ࡣࠊNF䃙B ࡣ୺࡟እ⭷ࠊෆ⭷ࢆ୰ᚰ࡟Ⓨ⌧ࡋ࡚࠸ࡓࠋ  ࣛࢵࢺ኱ື⬦⒗ࣔࢹ࡛ࣝࡣࠊ඲࡚ࣈࢱ⮅࢚ࣛࢫࢱ࣮ࢮฎ⌮࡟ࡼࡾࠊ㸰㐌㛫┠࡟ࡣ඲࡚᭷ព࡞⒗ᚄࡢ ᣑ኱ࡀⓎ⏕ࡋࡓࠋࢹࢥ࢖ࡢᑟධࡣࠊFITC ࣛ࣋ࣝࢹࢥ࢖࡟ࡼࡾ⭡㒊኱ື⬦ࡢእ⭷ࡸ୰⭷࡟Ꮡᅾࡋࠊࢩ ࣮ࢺᅛᐃᚋ㸵᪥ࡲ࡛☜ㄆࡋࡓࠋ  ศᏊ⏕≀Ꮫⓗホ౯࡟࠾࠸࡚ࠊ࣓࢟ࣛࢹࢥ࢖࡛ࡣࠊࢫࢡࣛࣥࣈࣝ࡜ẚ㍑ࡋ࡚ࠊ኱ື⬦⒗ቨ࡛ࡢ NF䃙B ࡜ ets-1 ࡀྠ᫬࡟ᢚไࡉࢀࠊࡑࡋ࡚ࠊྠᵝ࡟ࠊ኱ື⬦⒗ቨ࡛ࡢ MMP-9 ࡢⓎ⌧ࡸάᛶࡶᢚไࡉࢀࡓࠋ  ⑓⌮⤌⧊Ꮫⓗホ౯࡟࠾࠸࡚ࠊ࣓࢟ࣛࢹࢥ࢖࡛ࡣࠊࢫࢡࣛࣥࣈࣝ࡜ẚ㍑ࡋ࡚ࠊ኱ື⬦ቨࡢᙎᛶ⥺⥔ࡢ ⺮ⓑ⼥ゎࡀῶᑡࡋࠊࡲࡓࠊ኱ື⬦⒗ቨ࡛ࡢ࣐ࢡࣟࣇ࢓࣮ࢪᩘࡶῶᑡࡋࡓࠋ  ᙧែⓗホ౯࡟࠾࠸࡚ࠊ⾡ᚋ㸲㐌ࡲ࡛ࡢ኱ື⬦⒗ᚄࡢᢚไຠᯝࢆ㸲⩌࡛ẚ㍑ࡋࡓࠋ࣓࢟ࣛࢹࢥ࢖⩌࡜ NF䃙B ࢹࢥ࢖⩌ࡢ㸰⩌ࡣࠊ㠀ࢹࢥ࢖⩌ࡸࢫࢡࣛࣥࣈࣝࡢ㸰⩌ࡼࡾ᭷ព࡟⒗ᚄᣑ኱ࡀᢚไࡉࢀࡓࠋࡑࡋ ࡚ࠊ࣓࢟ࣛࢹࢥ࢖⩌ࡣࠊNF䃙B ࢹࢥ࢖⩌ࡼࡾ᭷ព࡟⒗ᚄᣑ኱ࡀᢚไࡉࢀࡓࠋ  ⪃ ᐹ  ⌧ᅾࠊ኱ື⬦⒗ࡢⓎ⏕ࡸ㐍ᒎࢆ᏶඲࡟㜼Ṇ࡛ࡁࡿ⸆≀἞⒪ࡣᏑᅾࡋ࡞࠸ࠋࡋ࠿ࡋࠊ኱ື⬦⒗ࡢ⑓ែ ࡀ኱ື⬦ቨ࡟࠾ࡅࡿ࢚ࣛࢫࢳࣥࡢ◚ቯ࡛࠶ࡾࠊࡑࡢ୺࡞ཎᅉ࡜ࡋ࡚ MMPs άᛶࡢஹ㐍ࡀド᫂ࡉࢀ࡚ ࠸ࡿࡇ࡜࠿ࡽࠊMMPs άᛶࢆᢚไࡍࡿࡇ࡜࡟ࡼࡾ᪂ࡋ࠸⸆≀἞⒪ࡀᐇ⌧࡛ࡁࡿ࡜⪃࠼ࡽࢀࡿࠋ௒ᅇࠊ ᪂ࡋ࠸㑇ఏᏊ἞⒪࡜ࡋ࡚ࠊ࠾࡜ࡾᆺ᰾㓟་⸆㸦ࢹࢥ࢖㸧ࢆ⏝࠸࡚ࠊMMPs ࡢ㌿෗ㄪ⠇ᅉᏊ࡛࠶ࡿ NF 䃙B ࡜ ets ࢆྠ᫬࡟ᢚไࡍࡿ࣓࢟ࣛࢹࢥ࢖ࢆ⪃᱌ࡋࡓࠋ ࣄࢺ┿ᛶ኱ື⬦⒗࡜ᐇ㦂ⓗ኱ື⬦⒗ࡢ┦㐪ࢆ᳨ウࡍࡿࡓࡵࠊࣄࢺ⭡㒊኱ື⬦⒗ቨ࡟࠾ࡅࡿ NF䃙B άᛶࡢஹ㐍ࢆ☜ㄆࡋࠊࡑࡋ࡚సᡂࡋࡓࣛࢵࢺ኱ື⬦⒗ࣔࢹࣝ࡟࠾࠸࡚ࡶ NF䃙B ࡜ ets-1 άᛶࡢஹ㐍ࠊ ୪ࡧ࡟ MMP-9 Ⓨ⌧ࡸάᛶࡢஹ㐍ࢆド᫂ࡋࡓࠋࡇࢀࡽࡢ⤖ᯝ࠿ࡽࠊࣛࢵࢺ኱ື⬦⒗ࣔࢹࣝ࡟࠾࠸࡚ࡶࠊ ࣄࢺ⭡㒊኱ື⬦⒗࡜ྠᵝ࡟ NF䃙B ࡸ ets-1 ࡢάᛶஹ㐍࡟ࡼࡿ MMPs ࡢ㐣๫Ⓨ⌧ࡀ኱ື⬦⒗ࡢᙧᡂ࡟ 㛵୚ࡍࡿ࡜⪃࠼ࡽࢀࡿࠋ  ࡇࡢࣛࢵࢺ኱ື⬦⒗ࣔࢹࣝ࡟࠾࠸࡚ࠊ࣓࢟ࣛࢹࢥ࢖ࡢຠᯝࢆ NF䃙B ࢹࢥ࢖ࡸࢫࢡࣛࣥࣈࣝࢹࢥ࢖ ࡜ẚ㍑ࡋࡓࠋ࣓࢟ࣛࢹࢥ࢖࡛ࡣࠊ⒗ᚄᣑ኱ࡢᢚไࠊ⒗ቨ NF䃙B ࡜ ets-1 ࡢ㌿෗άᛶࡢᢚไࠊ୪ࡧ࡟⒗ ቨ MMPs ࡢⓎ⌧࡜άᛶࡢᢚไࡀド᫂ࡉࢀࠊ⸆≀἞⒪࡜ࡋ࡚ࡢྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋࡲࡓ⒗ቨෆ࣐ࢡ ࣟࣇ࢓࣮ࢪᩘࡢῶᑡࡀ࠶ࡾࠊ࣐ࢡࣟࣇ࢓࣮ࢪࡢ㐟㉮ࡶᢚไࡍࡿࡇ࡜ࡀ᥎ᐹࡉࢀࡓࠋࡇ࠺ࡋࡓ⤖ᯝ࠿ࡽࠊ ࣓࢟ࣛࢹࢥ࢖ࡢ MMPs ࡢᢚไຠᯝࡣࠊNF䃙B ࡜ ets-1 ࡢ㌿෗ᢚไࠊ࣐ࢡࣟࣇ࢓࣮ࢪࡸ⾑⟶ᖹ⁥➽⣽ ⬊࡞࡝ࡢ⏘⏕⣽⬊࠿ࡽࡢศἪᢚไࠊ࣐ࢡࣟࣇ࢓࣮ࢪࡢ㐟㉮ᢚไ࡟ࡼࡿ⥲ྜⓗస⏝࡜⪃࠼ࡽࢀࡿࠋ௨ୖ ࡢࡼ࠺࡞᳨ウ࠿ࡽࠊNF䃙B ࡜ ets ࢆྠ᫬࡟ᢚไࡍࡿ࣓࢟ࣛࢹࢥ࢖ࡣࠊ኱ື⬦⒗ࡢⓎ⏕ࡸ㐍ᒎࢆᢚไࡍ ࡿ᪂ࡋ࠸㑇ఏᏊ἞⒪࡜࡞ࡿྍ⬟ᛶࡀ࠶ࡿࠋ  ⤖ ㄽ  ㌿෗ㄪ⠇ᅉᏊ࡛࠶ࡿ NF䃙B ࡜ ets ࢆྠ᫬࡟ᢚไࡍࡿ࣓࢟ࣛࢹࢥ࢖ࡣࠊᐇ㦂ⓗ኱ື⬦⒗ࡢ㐍ᒎࢆᢚ ไࡋࡓࡇ࡜࠿ࡽࠊ኱ື⬦⒗ࡢ᪂ࡋ࠸㑇ఏᏊ἞⒪࡜࡞ࡿྍ⬟ᛶࡀ࠶ࡿࠋ኱ື⬦⒗ࡢ㐍ᒎ࡟ࡣࠊNF䃙B ࡜ etsࡀ㔜せ࡞ᙺ๭ࢆ₇ࡎࡿ࡜⪃࠼ࡽࢀࡿࠋ

ㄽ ᩥ ᑂ ᰝ ࡢ ⤖ ᯝ ࡢ せ ᪨

 ᮏ◊✲ࡣࠊࣛࢵࢺ኱ື⬦⒗ࣔࢹࣝࢆ⏝࠸࡚ࠊ⒗ቨ MMPs ࡢᢚไ࡟ࡼࡿ᪂ࡋ࠸㑇ఏᏊ἞⒪ࡢྍ⬟ᛶ ࢆ᳨ウࡋࡓࡶࡢ࡛࠶ࡿࠋ  MMPs ࡢ㌿෗ㄪ⠇ᅉᏊ࡛࠶ࡿ NF䃙B ࡜ ets-1 ࢆྠ᫬࡟ᢚไࡍࡿ࣓࢟ࣛࢹࢥ࢖ࢆ☜❧ࡋࡓࠋࡇࡢ࢟ ࣓ࣛࢹࢥ࢖ࢆࣛࢵࢺ኱ື⬦⒗ࣔࢹࣝ࡟ᑟධࡍࡿࡇ࡜࡟ࡼࡾࠊ⒗ᚄࡢᣑ኱ᢚไࠊ⒗ቨ NF䃙B ࡜ ets-1 ࡢ㌿෗ᢚไࠊ୪ࡧ࡟⒗ቨ MMPs ࡢⓎ⌧ᢚไࢆド᫂ࡋࡓࠋ  ࡇࢀࡽࡢෆᐜࡣࠊ┿ᛶ኱ື⬦⒗࡟ᑐࡍࡿ᪂ࡋ࠸㑇ఏᏊ἞⒪ࡢྍ⬟ᛶࢆ♧ࡋࡓࡶࡢ࡛࠶ࡾࠊ᫂ࡽ࠿࡟ Ꮫ⾡ࡢỈ‽ࢆ㧗ࡵࡓࡶࡢ࡜ㄆࡵࡿࠋ