Title アーティチョーク抽出物から得られたシナロピクリンのNF-κB転写活性阻害による光老化抑制効果( 本文(Fulltext) ) Author(s) 田中, 友香 Report No.(Doctoral Degree) 博士(薬科学) 連創博甲第25号 Issue Date 2014-03-25 Type 博士論文 Version ETD URL http://hdl.handle.net/20.500.12099/49053 ※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。
࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࡽᚓࡽࢀࡓ
ࢩࢼࣟࣆࢡࣜࣥࡢ
NF-
κB ㌿άᛶ㜼ᐖࡼࡿ
ග⪁ᢚไຠᯝ
Cynaropicrin from Cynara scolymus L. suppresses photoaging of skin
by inhibiting the transcription activity of nuclear factor- kappa B
2014
┠ḟ ┠ḟ ┠ḟ ┠ḟ ⥴ゝ 3 ヨ㦂ᮦᩱヨ㦂᪉ἲ 1. ᮦᩱヨ⸆ 13 2. ⣽⬊ᇵ㣴 13 3. 㑇ఏᏊᑟධࣝࢩࣇ࢙࣮ࣛࢮࢵࢭ 14 4. bFGF MMP-1 ࡢᐃ㔞 14 5. ࣓ࣛࣀࢧࢺቑṪ ᐃ 15 6. චᰁⰍࡼࡿ p65 ᒁᅾࡢほᐹ 15 7. UVB ↷ᑕ⓶ࣔࢹ࣐ࣝ࢘ࢫ 15 8. ⤫ィゎᯒ 17 ⤖ᯝ 1. NF-κB ㌿άᛶ㜼ᐖస⏝ 17 2. ࢩࢼࣟࣆࢡࣜࣥࡼࡿ NF-κB ࡢస⏝ᶵᡤ 20 3. bFGF ࠾ࡼࡧ MMP-1 ⏘⏕࠾ࡅࡿ⏘⏕㜼ᐖస⏝ 21 4. ࣓ࣛࣀࢧࢺቑṪ࠾ࡅࡿ㜼ᐖຠᯝ 22 5. ࢩࢼࣟࣆࢡࣜࣥࡢ࣐࢘ࢫ⤒ཱྀᢞࡼࡿ⾲⓶⫧ཌ࠾ࡼࡧ࣓ࣛࢽࣥ⏘⏕ࡢᢚไຠᯝ 23 ⪃ᐹ 27 ⤖ㄽ 28
ㅰ㎡ 28 ཧ⪃ᩥ⊩ 29 ཧ⪃㈨ᩱ࣭⥲ㄝ 35
⥴ゝ ⥴ゝ ⥴ゝ ⥴ゝ 㔝⳯ࡸᯝ≀ࡀ㇏ᐩ࡞㣗ࡣᗣ⥔ᣢḞࡏ࡞࠸ࡇࡣᗈࡃㄆ▱ࡉࢀ࡚࠸ࡿࠋཌ⏕ປാ ┬ࡢ㐠ႠࡍࡿWeb ࢧࢺࠊe-࣊ࣝࢫࢿࢵࢺ࡛ࡶ㔝⳯ࡢ᭷┈ᛶࡘ࠸࡚⤂ࡋࠊ⏕ά⩦័ ࢆண㜵ࡋࠊᗣ⥔ᣢࡢࡓࡵ㔝⳯㢮ࡢ1 ᪥ 350g ௨ୖࡢᦤྲྀࢆ᥎ዡࡋ࡚࠸ࡿࠋࡇࡢᵝ࡞㔝⳯ ࡸᯝ≀ࡢ᭷ຠᛶࡣ᳜≀ࡢ≉␗ⓗ࡞ᡂศ࡛࠶ࡿphytochemicals ࡼࡿࡶࡢࡉࢀ࡚࠸ࡿ1ࠋ Ꮫㄪᰝࡽࡶphytochemicals ࢆྵࡴ㣗ࡼࡾ⒴ࡸ⅖ᛶᝈࠊᚰ⾑⟶ᝈ࡞ࡢ៏ᛶ㞀 ᐖࢆᢚไࡋࠊࣝࢶࣁ࣐࣮ࡸࣃ࣮࢟ࣥࢯࣥࡢࣜࢫࢡࢆῶᑡࡉࡏࡿሗ࿌ࡉࢀ࡚࠸ࡿ1ࠋ Phytochemicals ࡣ᳜≀୰Ꮡᅾࡍࡿḟ௦ㅰ⏘≀ࡢࡇࢆ࠸࠺ࠋ᳜≀ࡀᙉ࠸⣸እ⥺ࡸẼೃࠊ ᐖࡸື≀࡞ࡢᵝࠎ࡞⎔ቃ࣭እᩛࡽ㌟ࢆᏲࡿࡓࡵ⮬ࡽసࡾฟࡋࡓᶵ⬟ᛶᡂศ࡛ࠊⰍ ࡸ㤶ࡾࠊ㎞ࠊⱞ࡞ྵࡲࢀࡿ㸦⾲㸯㸧2ࠋ୍⯡ⓗࡣ㏻ᖖࡢ㌟య⥔ᣢࡣᚲせࡉࢀ ࡞࠸ࡀᗣⰋ࠸ᙳ㡪ࢆ࠼ࡿྍ⬟ᛶࡀ࠶ࡿ᳜≀⏤᮶ࡢྜ≀ࡋ࡚ὀ┠ࡉࢀࠊ୍⯡㣗ရ ࡋ࡚㣗ࡉࢀࡿࡔࡅ࡛࡞ࡃࠊ⢝ရࡸᗣ㣗ရ࡞ከࡃ㓄ྜࡉࢀ࡚࠸ࡿࠋ࠼ࡤࠊࣈࢻ ࢘ࡢࣥࢺࢩࢽࣥࠊⲔࡢ࢝ࢸ࢟ࣥࠊࢽࣥࢪࣥࡸࡰࡕࡷࡢș㸫࢝ࣟࢸࣥࠊࢺ࣐ࢺࡢࣜࢥ ࣆࣥ࡞ࡀⰋࡃ▱ࡽࢀ࡚࠸ࡿࠋᮏ᮶ࠊphytochemicals ࡣࠊ᳜≀ࡀᐖ࡞ࡽ㌟ࢆᏲࡿࡓ ࡵ᳜≀ෆ࡛సࡾฟࡍࡶࡢ࡛ẅ⳦ຠᯝࡸᚷ㑊ຠᯝࢆ♧ࡍࠋࡋࡓࡀࡗ࡚㧗⏝㔞࡛ᦤྲྀࡋࡓሙ ྜࠊⓎ⒴≀㉁ࡸ⚄⤒ẘ࡞ࡾ࠺ࡿࡀࠊప⏝㔞࡛ᦤྲྀࡋࡓሙྜࠊ㐺ᛂྍ⬟࡞⣽⬊ࢫࢺࣞࢫᛂ ⟅⤒㊰ࢆάᛶࡋᗣ᭷┈࡞ാࡁࢆࡍࡿ1,2ࠋࡇࡢᵝ࡞⏝㔞ᛂࡌ࡚㏫ࡢຠᯝࢆᘬࡁ㉳ࡇ ࡍ⌧㇟ࡣ⏕≀Ꮫࡸ⸆࡛ᗈࡃほᐹࡉࢀ”hormesis” ゝࢃࢀ࡚࠸ࡿ1-3ࠋ Phytochemicals ࡢ hormesis ຠᯝࡼࡾࠊయࢆᵓᡂࡍࡿྛ⣽⬊ࡀࢫࢺࣞࢫ⪏࠼ࡿࡓࡵࡢᢠ
㓟㓝⣲ࠊࢱࣥࣃࢡ㉁ࢩࣕ࣌ࣟࣥࠊࣇ࢙࣮ࢬ-2 㓝⣲ࠊ⚄⤒ᰤ㣴ᅉᏊ࡞ࡢ⏘⏕ࢆಁ㐍ࡉࡏࠊ ⚄⤒⣽⬊ࡢಖㆤస⏝ࡸᢠ⒴స⏝ࠊᗣಁ㐍స⏝ࢆ♧ࡍࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ1,2ࠋ ⢝ရࡣ⸆ἲ➨2 ᮲➨ 3 㡯࠾࠸࡚ࠊࡲࡓࠊᣦᐃࡉࢀࡓ⠊ᅖෆ࡛ຠ⬟࣭ຠᯝࢆ⾲♧࡛ ࡁࡿ་⸆㒊እရࡣ⸆ἲ➨2 ᮲➨ 2 㡯࠾࠸࡚ᐃ⩏ࡉࢀ࡚࠸ࡿࠋ་⸆㒊እရࡢ࡞ヱᙜၟ ရࡣ⨾ⓑࡸ⑫⒔ண㜵ຠᯝࡀᮇᚅ࡛ࡁࡿ⸆⏝⢝ရࠊ⫱ẟࠊ࣮࣊࢝ࣛࠊṑ࿘࣭ṑ ண㜵ࡢṑࡳࡀࡁ࡞ࡀ࠶ࡿࠋࡉࡽࠊ2001 ᖺࡢ⢝ရࡢつไ⦆ࡼࡾࡑࢀࡲ࡛ཌ⏕ປാ ⮧ㄆྍࡉࢀࡓᡂศ௨እࡣ⢝ရ㓄ྜ࡛ࡁ࡞ࡗࡓࡀࠊᡂศ⾲♧ࢆ⩏ົࡅࡽࢀࡓ ࠺࠼࡛ࠊ⸆ἲ⢝ရᇶ‽࡛⚗Ṇ࣭ไ㝈ࡀ࠶ࡿᡂศࢆ㝖ࡁࠊྛᴗࡀᏳᛶࢆ☜ㄆࡋࡓ࠺ ࠼࡛⮬⏤㓄ྜࡍࡿࡇࡀྍ⬟࡞ࡗࡓࠋ୍᪉ࠊᗣ㣗ရࡣᗈࡃᗣࢆቑ㐍ࡉࡏࡿ┠ⓗ࡛ ㈍ࠊ⏝ࡉࢀࡿ㣗ရ⯡ࢆᣦࡋࠊἲᚊୖࠊᐃ⩏ࡣࡉࢀ࡚࠸࡞࠸ࠋࡑࡇ࡛ࠊ1991 ᖺᗣ ቑ㐍ἲࡼࡾ≉ᐃಖ⏝㣗ရࡀᐃ⩏ࡉࢀࠊᅜࡢᐃࡵࡓつ᱁ࡸᇶ‽ࢆ‶ࡓࡍ㣗ရࡘ࠸࡚ࡣ ಖᶵ⬟ࡢ⾲♧ࡀྍ⬟࡞ࡗࡓࠋࡇࡢᵝ࡞ὶࢀࡽ୍⯡ᾘ㈝⪅ࡣ⢝ရࡸᗣ㣗ရࡢᡂศ ࡢᶵ⬟ᛶࡘ࠸࡚┠ࡍࡿᶵࡀቑ࠼ࠊPhytochemicals ࡘ࠸࡚ࡶᵝࠎ࡞࣓ࢹ࡛⤂ ࡉࢀࠊㄆ▱ࡉࢀࡿࡼ࠺࡞ࡗ࡚࠸ࡿࠋ࠼ࡤࠊ་⸆㒊እရࡢ⨾ⓑᡂศ࡛࠶ࡿࣝࣈࢳࣥࡣ ࢥࢣࣔࣔࡸ࢘࣡࢘ࣝࢩ⏤᮶ࡢࣇ࢙ࣀ࣮ࣝᛶ㓄⢾య࡛࣓ࣛࢽࣥࡢྜᡂ㛵ࢃࡿ㓝⣲ࠊࢳࣟࢩ ࢼ࣮ࢮࡢ㜼ᐖࡼࡾ⨾ⓑస⏝ࢆ♧ࡍࠋࡲࡓࠊすὒࣁ࣮ࣈ࡛࠶ࡿ࣑࣮࢝ࣔࣝࡽᢳฟࡉࢀࡓ ࣑࢝ࣔࣛET ࡶ」ᩘࡢ Phytochemicals ࢆ᭷ຠᡂศࡍࡿ⨾ⓑᡂศࡋ࡚ᐇ⏝ࡉࢀ࡚࠸ࡿࠋ ࡲࡓࠊ2011 ᖺࡣࠊ་⸆㒊እရ௨እࡢ୍⯡⢝ရ࠾࠸࡚ࢩ࣡㛵ࡍࡿຠᯝࢆㅻ࠺ࡇࡀ ㄆྍࡉࢀࡓࠋ᪥ᮏࢆྵࡵୡ⏺࡛⮬⏕ࡍࡿࣄ࣓ࣇ࢘ࣟࡽᢳฟࡉࢀࡿᡂศࡣࠊ⣸እ⥺ࡼࡾ άᛶࡍࡿ࣐ࢫࢺࢭࣝ⏤᮶ࡢ㓝⣲ࠊࢺࣜࣉࢱ࣮ࢮࢆ㜼ᐖࡍࡿࠋࢺࣜࣉࢱ࣮ࢮࡣࠊ⣽⬊እ࣐
ࢺࣜࢵࢡࢫࡸᇶᗏ⭷ࢆศゎࡋࠊග⪁㛵ಀࡍࡿࡉࢀ࡚࠸ࡿ4ࠋࡋࡓࡀࡗ࡚ࠊࣄ࣓ࣇ࢘ ࣟᡂศࡣ⣸እ⥺ࡼࡾᘬࡁ㉳ࡇࡉࢀࡿࢺࣜࣉࢱ࣮ࢮࢆ㜼ᐖࡋࠊࢥ࣮ࣛࢤࣥࡸᇶᗏ⭷ࡢศゎ ࡼࡿࢩ࣡ࡢᙧᡂࢆᢚไࡍࡿග⪁ᢚไస⏝ࢆ♧ࡍࠋ㇋ࡢࢯࣇࣛ࣎ࣥࡣዪᛶ࣍ࣝࣔࣥ ࡛࠶ࡿ࢚ࢫࢺࣟࢤࣥࣞࢭࣉࢱ࣮⤖ྜࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠾ࡾࠊ࣍ࣝࣔࣥᵝάᛶࢆ᭷ࡍ ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ5ࠋዪᛶࡣ᭦ᖺᮇධࡿዪᛶ࣍ࣝࣔࣥ⏘⏕㔞ࡀపୗࡋࠊ᭦ᖺᮇ㞀 ᐖࢆᘬࡁ㉳ࡇࡍAࠋࡇࡢ㝿ࠊ⓶࠾࠸࡚ࡶ┿⓶ᵓᡂᡂศ࡛࠶ࡿࢥ࣮ࣛࢤࣥࡸ࢚ࣛࢫࢳࣥ ࡢ᪂⏕ࡀῶᑡࡍࡿࡇ࡛ࢩ࣡ࡸࡓࡿࡳࡢཎᅉ࡞ࡿࠋ㇋ࢯࣇࣛ࣎ࣥࡣዪᛶ࣍ࣝࣔࣥᵝ άᛶࢆ♧ࡍࡇࡽࠊᨃఝⓗ࣍ࣝࣔࣥࡢ㊊ࢆ⿵࠸⨾ᐜຠᯝࢆ♧ࡍࠋࢥ࢞ࢿࣂࢼྵࡲ ࢀࡿࣂ࢝ࣜࣥࠊࣂ࢝ࣞࣥࡣᙉ࠸ᢠ㓟స⏝ࡀㄆࡵࡽࢀࠊάᛶ㓟⣲ࡼࡿ⣽⬊㞀ᐖ ࢆ㍍ῶࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ6ࠋάᛶ㓟⣲ࡣ㌟యࢆㆤࡿᶵᵓࡢࡦࡘ࡛ࡣ࠶ࡿࡀࠊ㐣 ࡞άᛶ㓟⣲ࡣᚲせ௨ୖ㌟యࢆᵓᡂࡍࡿ⣽⬊ࡸྛ✀⺮ⓑ㉁➼ࢆ◚ቯࡍࡿࡇࡀ▱ࡽࢀ࡚࠾ ࡾࠊࡇࢀࡀ⪁ࡸࡢཎᅉၐ࠼ࡿᏛ⪅ࡶ࠸ࡿAࠋ≉⣸እ⥺ࡣ⓶ෆ࡛ࡢάᛶ㓟⣲ࡢ ቑຍࢆᘬࡁࡇࡍࡓࡵࠊάᛶ㓟⣲ࡼࡿ㞀ᐖࢆ㏿ࡸண㜵ᨵၿࡍࡿᚲせࡀ࠶ࡿࠋࡇࡢᢠ㓟 ᶵ⬟ࢆྲྀࡾධࢀࡿࡓࡵࠊ┤᥋⫙ሬᕸࡍࡿࡇࡀྍ⬟࡞⢝ရࡣ᭷ຠ࡞ᡭẁࡢࡦࡘ ࡛࠶ࡾࠊࡑࡢ㟂せࡀ㧗ࡲࡗ࡚࠸ࡿࠋࡑࢀ௨እࡶPhytochemicals ࡼࡿ hormesis ຠᯝࢆᛂ ⏝ࡋ࡚࠸ࡿ⪃࠼ࡽࢀࡿ᳜≀࢚࢟ࢫࡣᩘⓒ✀㢮ࡶཬࡧࠊከࡃࡢ⢝ရࠊ་⸆㒊እရࠊ ᗣ㣗ရῧຍࡉࢀ࡚࠸ࡿࠋ 2012 ᖺㄪᰝ♫ࣥࢸ࣮ࢪࡼࡿ 10~70 ௦ࡢ⏨ዪᑐ㇟ࡢㄪᰝ࡛ࡣᗣ㣗ရ࣭ࢧࣉࣜ ࣓ࣥࢺࡢᕷሙつᶍࡣ1 4 ༓൨ࡶࡢࡰࡿࠋᗣ㣗ရࡣࠊ⨾⫙࣭⫙ࢣࠊᗣ⥔ᣢࠊ 㛵⠇ࡢᗣ⥔ᣢࠊῶ㔞࡞ᵝࠎ࡞࢝ࢸࢦ࣮ࣜࡀᏑᅾࡍࡿࡀࠊ᭱ࡶࡁ࡞ᕷሙࡣ⨾⫙࣭⫙ࢣ
࡛ࡑࡢつᶍࡣ1,674 ൨ࠊࡘ࠸࡛ᗣ⥔ᣢ࣭ቑ㐍ࡢ 1,450 ൨⥆ࡃࠋࡇࢀࡣ⏨ᛶࡼࡾࡶ ዪᛶࠊ≉40 ௦௨ୖࡢዪᛶࡢ⏝⪅ࡀከ࠸ࡇࡽࣥࢳ࢚ࢪࣥࢢ࠸࠺ゝⴥࡶ ᗣ㣗ရ࡛ࡶ⨾ᐜ㛵ࡍࡿὀ┠ᗘࡀ㧗࠸ࡓࡵᛮࢃࢀࡿࠋ ⣸እ⥺ࡣࠊ⓶ࡢග⪁ࢆᘬࡁ㉳ࡇࡍ᭷ᐖ࡞⎔ቃせᅉࡢࡦࡘ࡛࠶ࡿ7ࠋග⪁ࡣ៏ᛶ ⓗ࡞⣸እ⥺↷ᑕࡼࡗ࡚⓶ࡀ᪩ᮇ⪁ࡋࡓ≧ែ࡛ࠊ⩦័ⓗኴ㝧ගࢆᾎࡧࡓ⪁ேࡢ⓶ ࡣ⾰᭹࡛そࢃࢀࡓ㒊ศ㟢ฟࡋࡓ㒊ศࢆẚ㍑ࡋࡓሙྜࠊ㟢ฟ㒊ศࡢ᪉ࡀࡼࡾ⪁ࡋ࡚ぢ ࠼ࡿ8ࠋ୍⯡ᖺࢆ㔜ࡡࡿࡈᖺ┦ᛂࡢ㢦❧ࡕ࡞ࡗ࡚ࡃࡿࡀࠊ↓ព㆑ࡇࡢุ᩿ᇶ‽ ࡞ࡗ࡚࠸ࡿࡢࡀࠊࢩ࣡ࠊࡓࡿࡳࠊࢩ࣑࡛࠶ࡿAࠋࡇࡢኚࡢ≉ᚩࡣࠊே✀ࡈ␗࡞ࡿࠋ ࠼ࡤࠊ࣓ࣛࢽࣥⰍ⣲ࡢከ࠸㯮ேࡣ⣸እ⥺⪏ᛶࡀ㧗ࡃࠊࢩ࣡࡞ࡀᙧᡂࡉࢀࡃ࠸ࠋ୍᪉ࠊ ࣓ࣛࢽࣥⰍ⣲ࡢᑡ࡞࠸ⓑேࡣẚ㍑ⓗ┿⓶ࡢࢲ࣓࣮ࢪࡀ㧗ࡃ࡞ࡿࡇ࡛ࠊࢩ࣡ࡸࡓࡿࡳࡀ ࠾ࡁࡸࡍ࠸ࠋࡑࡢ୰㛫࡛࠶ࡿ㯤Ⰽே✀ࡣࠊ㐺ᗘ⣸እ⥺㜵ᚚࡍࡿࡓࡵࢩ࣡ࡸࡓࡿࡳࡣⓑே ẚ࡚ᑡ࡞࠸ࡀࠊ࣓ࣛࢽࣥⰍ⣲ࡢ␗ᖖ⏘⏕ࡼࡿࢩ࣑ࡀࡼࡾ┠❧ࡕࡸࡍ࠸ഴྥࡀ࠶ࡿࠋ ࡲࡓࠊ⣸እ⥺ࡢᙉ࠸࣮࢜ࢫࢺ࡛ࣛࣜࡣᗣ⿕ᐖࢆண㜵ࡍࡿ┠ⓗ࡛ࠊᅜ⟇ࡋ࡚ ᖺ௦ ࠕࢫࣜࢵࣉࠊࢫࣟࢵࣉࠊࢫࣛࢵࣉࠊࣛࢵࣉ㸦6OLS6ORS6ODS:UDS㸧ࠖࣉࣟࢢ࣒ࣛࡀᑟධ ࡉࢀ࡚࠸ࡿࠋࡇࢀࡣࠊ㛗ࡑ࡛ࢆ╔ࡿࠊ᪥↝ࡅṆࡵࢆሬࡿࠊᖗᏊࢆࡪࡿࠊࢧࣥࢢࣛࢫࢆࡍ ࡿࡇࢆ᥎ዡࡍࡿࡶࡢ࡛࠶ࡾࠊ⣸እ⥺ࡢᗣ⿕ᐖࡀ῝้࡞ၥ㢟࡛࠶ࡿࡇࢆ≀ㄒࡗ࡚࠸ࡿࠋ ᪥ᮏ࠾࠸࡚ࡶࠊࡘ࡚ẕᏊᡭᖒࡣ᪥ගᾎࡀ᥎ዡࡍࡿグ㏙ࡀ࠶ࡗࡓࡀࠊ ᖺࡼࡾ๐㝖 ࡉࢀ࡚࠸ࡿࠋ᪥ගᾎࡣࣅࢱ࣑ࣥ ' ࢆయෆ࡛సࡿࡓࡵᚲせࡉࢀ࡚࠸ࡓࡀࠊ⌧ᅾࡣ㏻ᖖࡢ 㣗࡛౪⤥ྍ⬟࡛࠶ࡿࡓࡵࡑࡢᜠᜨࡣᑡ࡞࠸ุ᩿ࡉࢀࡓࠋ ᅇࠊᡃࠎࡣ⣸እ⥺ࡼࡗ࡚ࡶࡓࡽࡉࢀࡿ⓶⪁ࠊࡍ࡞ࢃࡕ⓶ග⪁╔┠ࡋࡓࠋ
⣸እ⥺ࡣࠊࣥࢱ࣮ࣟ࢟ࣥ-1㸦IL-1㸧ࡸ⭘⒆ቯṚᅉᏊࣝࣇ㸦TNF-α㸧࡞ࡢࢧࢺ࢝ ࣥࢆศἪࡉࡏࠊ⅖ࣉࣟࢭࢫࢆㄏᑟࡍࡿࠋࡑࡢ⤖ᯝࠊ⣸እ⥺ࢆ↷ᑕࡉࢀࡓ⓶ࡣࠊ⓾ࡸ ࣓࢟ࡢࢀࠊ⇱ࠊࡓࡿࡳ࡞ࡢ≉ᚩⓗእほࡀㄆࡵࡽࢀࡿ8,9ࠋ⾲2 ♧ࡋࡓ࠾ࡾຍ㱋 ࡼࡿ⮬↛⪁ග⪁ࡣእほࡢ᫂ࡽ࡞㐪࠸ࡔࡅ࡛࡞ࡃࢣࣛࢳࣀࢧࢺࡸ࣓ࣛࣀࢧࢺ ࡞ࡢ⣽⬊ࡸ⏘⏕ࡉࢀࡿᡂศࡶ㐪࠸ࡀㄆࡵࡽࢀࡿBࠋࡇࢀࡽࡢኚࡣࠊࢣࣛࢳࣀࢧ ࢺࡸ࣓ࣛࣀࢧࢺࡢ㐣ቑṪࢥ࣮ࣛࢤࣥ⥺⥔ࡢศゎࡀ㛵ࡋ࡚࠸ࡿ10,11ࠋࡇࡢ⓶ග⪁ ࡢⓎ⏕ᶵᗎࠊ㌿ᅉᏊNuclear factor κB 㸦NF-κB㸧ࡀ῝ࡃ㛵ࡋ࡚࠸ࡿࡇࡀ㏆ᖺሗ࿌ ࡉࢀ࡚࠸ࡿ8,9ࠋNF-κB ࡣࠊ⅖㛵ࢃࡿ㔜せ࡞㌿ᅉᏊࡋ࡚ࠊ㛗ᖺࢃࡓࡾ་⒪ࡢศ㔝 ࡛◊✲ࡀ㔜ࡡࡽࢀ࡚ࡁࡓࠋࡓ࠼ࡤࠊNF-κB ࡣࡀࢇࡸ࣐ࣜ࢘ࢳࡢែ㐍⾜㛵ࡋ࡚࠾ࡾ 12,13ࠊࡑࡢ㜼ᐖࡢ㛤Ⓨࡀୡ⏺୰ࡢ◊✲ᶵ㛵࡛࡞ࡉࢀ࡚࠸ࡿࠋNF-κB ࡑࡢࡶࡢࡣࠊ⏕ά ືᚲ㡲࡞㌿ᅉᏊ࡛ࡣ࠶ࡿࡀࠊࡢ㌿ᅉᏊẚࠊከࡃࡢࡘ࡞ࡀࡿ⅖ᛂ 㛵ࡍࡿ㑇ఏᏊⓎ⌧ࢆࡼࡾከࡃ㌿ࡍࡿࡇࡽࠊண㜵ᨵၿ⸆ࡋ࡚ࡢࢱ࣮ࢤࢵࢺ࡞ࡾ ࡸࡍ࠸⫼ᬒࡀ࠶ࡿ14ࠋ⓶ศ㔝㛵ࡋ࡚ࡶࠊࢺࣆ࣮ᛶ⓶⅖ࡸⓄNF-κB ࡀ῝ࡃ㛵 ࡋ࡚࠾ࡾ12ࠊࡢྠᵝ◊✲ࡀ࡞ࡉࢀ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽࠊ⮳ࡽ࡞࠸㌟య ࡢኚᑐࡍࡿNF-κB ࡢᙳ㡪ࡘ࠸࡚ࡢ▱ぢࡣᑡ࡞࠸ࠋࡓ࠼ࡤࠊ⪁࠸࠺⏕⌮⌧㇟ࡣ ☜ᐇ㌟యࡢ⤌ࡳࢆኚࡉࡏᚓࡿ⌧㇟࡛࠶ࡿࡀࠊ࡛ࡣ࡞࠸ࡓࡵ✚ᴟⓗ◊✲ࡀ࡞ࡉ ࢀ࡚ࡇ࡞ࡗࡓࠋࡑࡢࡼ࠺࡞⫼ᬒࡢ࡞ࠊ⣸እ⥺ࡼࡿ⓶⪁NF-κB ࡢ㛵㐃ࡘ࠸ ࡚ሗ࿌ࡀ࡞ࡉࢀࠊ⣸እ⥺ࡼࡗ࡚άᛶࡋࡓ㐣࡞NF-κB ࡀ⓶⪁ࢆಁ㐍ࡍࡿࡇࡀ࿘ ▱ࡉࢀࡓ8,9ࠋࡋࡋࠊ᪤Ꮡࡢᗣ㣗ရࡸ⢝ရࠊ་⸆㒊እရ࠾࠸࡚ࡶᢠග⪁ຠᯝࢆㅻ ࡗࡓࡶࡢࡣ࠶ࡿࡀNF-κB ࢆࢱ࣮ࢤࢵࢺࡋࡓ〇ရࡣ࡞ࡗࡓࠋ
⓶⣽⬊ෆ࡛⣸እ⥺ࡸ⅖ᛶࢧࢺ࢝ࣥ࡞ࡢ่⃭ࡼࡾࠊNF-κB ࡀάᛶࡋࠊ࣐ ࢺࣜࢵࢡࢫ࣭࣓ࢱࣟࣉࣟࢸ࣮ࢮ-1㸦MMP-1㸧ࡀ⏘⏕ࡉࢀࠊࡑࡢ⤖ᯝࠊࢥ࣮ࣛࢤࣥ⥺⥔ࡢ ◚ቯࡀ㉳ࡁࡿ15,16ࠋࡲࡓNF-κB ࡣࠊሷᇶᛶ⥺⥔ⱆ⣽⬊ᡂ㛗ᅉᏊ㸦bFGF㸧ࡢ⏘⏕ࢆಁ㐍ࡍࡿ ࡇࡶሗ࿌ࡉࢀࠊࢣࣛࢳࣀࢧࢺࡸ࣓ࣛࣀࢧࢺࡢ㐣ቑṪࡢཎᅉ࡞ࡿ17,18ࠋࡉࡽࠊ⣸ እ⥺↷ᑕࡼࡿNF-κB άᛶࢆ㏻ࡋ࡚ bFGF ⏘⏕ࡀㄏⓎࡉࢀࡿࡇࡶሗ࿌ࡉࢀ࡚࠸ࡿ19,20ࠋ NF-κB ࡣಶูࡢࢧࣈࣘࢽࢵࢺ࡛࠶ࡿ 5 ࡘࡢ⊂❧ࡋࡓ㑇ఏᏊ NFKB1㸦p105㸧ࠊNFKB2㸦p100㸧ࠊ RelA㸦p65㸧ࠊc-rel ཬࡧ relB ࡽᡂࡿ21ࠋ࣍ࣔཪࡣ࣊ࢸࣟࢲ࣐࣮ࡽ࡞ࡿNF-κB ࢱࣥࣃ ࢡ㉁ࡣᶆⓗ㑇ఏᏊࡢ⤖ྜ㒊࡛DNA 㓄ิ⤖ྜࡍࡿࡇ࡛㌿ࢆㄪ⠇ࡍࡿ22ࠋ NF-κB ࡣ㏻ᖖࠊᢚไศᏊࡋ࡚▱ࡽࢀࡿ inhibitor κB 㸦IκB㸧⤖ྜࡋάᛶ」ྜయࡋ ࡚⣽⬊㉁␃ࡲࡿࠋࡋࡋࠊIL-1 ࡸ TNF-αࡢࡼ࠺࡞ࢧࢺ࢝ࣥࡼࡾࣜࣥ㓟ࠊࣘࣅ࢟ ࢳࣥࡉࢀࡓᚋࠊIκB ࢱࣥࣃࢡ㉁ࡀศゎࡉࢀ NF-κB ࡀ᰾⛣⾜ࡍࡿ23-25ࠋࡉࡽNF-κB ࡢ άᛶࡼࡾග⪁ࡀᘬࡁ㉳ࡇࡉࢀࡓࠊࡑࡢ㜼ᐖ࡛࠶ࡿparthenolide ࢆ⭡⭍ෆᢞࡋࡓ ࡇࢁࠊ⣸እ⥺↷ᑕࡼࡿbFGF ࡸ MMP-1 ࡢ⏘⏕ࢆ㜼ᐖࡋࠊ⾲⓶ゅ⣽⬊ࡸ࣓ࣛࣀࢧࢺ ࡢ㐣ቑṪࢆᢚไࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ17,18,23,26ࠋParthenolide ࡣ࢟ࢡ⛉ࢼࢶࢩࣟࢠࢡ ྵࡲࢀࡿࢭࢫ࢟ࢸࣝ࣌ࣥࣛࢡࢺࣥࡢ1 ✀࡛࠶ࡿࠋࢭࢫ࢟ࢸࣝ࣌ࣥࣛࢡࢺࣥࡣ⅖ᛶᝈࡢ ⒪⏝ࡉࢀࡿ⸆⏝᳜≀ࡢ᭷ຠᡂศࡋ࡚▱ࡽࢀࠊα−࣓ࢳࣞࣥ−γ−ࣛࢡࢺࣥ⎔ࡀᣢࡘࢳ ࣮࢜ࣝᇶࡢᛂᛶࡼࡗ࡚⏕⌮άᛶࢆ♧ࡍࡇࡀ▱ࡽࢀ࡚࠸ࡿ27ࠋParthenolide ࡣࣜࣥ㓟 ࡉࢀࡓΙκB ࡢศゎࢆ㜼ᐖࡍࡿࡇ࡛ NF-κB 㜼ᐖάᛶࢆ♧ࡍࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ27ࠋ NF-κB ࡢ㜼ᐖࡣࠊ⅖ᛂࡢᝏᚠ⎔ࢆᢚไࡋࠊ⓶ࡢග⪁ࡢ㜵Ṇຠᯝⓗ࡞ാࡁࢆࡍ ࡿ㸦ᅗ1㸧ࠋParthenolide ௨እࡶኳ↛≀ࡢ୰ࡣ NF-κB 㜼ᐖάᛶࢆࡶࡘ᭷┈࡞ᡂศࡢᏑᅾ
ࡀᮇᚅࡉࢀࡓࠋ ᡃࠎࡣࠊTNF-α่⃭ࡼࡿ NF-κB ࡢ㌿άᛶ㜼ᐖࡢస⏝ࢆ 200 ௨ୖࡢ᳜≀࢚࢟ࢫ࡛ヨ 㦂ࡋࡓࠋࡑࡢ⤖ᯝࠊ࣮ࢸࢳ࣮ࣙࢡ㸦Cynara scolymus L.㸧ᙉ࠸ NF-κB ㌿άᛶ㜼ᐖ స⏝ࢆ☜ㄆࡋࡓ㸦ᅗ2㸧ࠋ ࣮ࢸࢳ࣮ࣙࢡࡣᆅ୰ᾏἢᓊ࠾ࡼࡧࣇࣜ࢝㒊ཎ⏘ࡢ࢟ࢡ⛉ࡢ᳜≀࡛ࠊ⣙2 ࣓࣮ࢺ ࣝࡢ㧗ࡉᡂ㛗ࡋࠊ㟷⣸Ⰽࡢⰼᘚࢆ᭷ࡍࡿࠋࡶࡶࡣᆅ୰ᾏ୰す㒊㔝⏕ࡋࠊⴥࢆ㣗 ⏝ࡋࡓ࢝ࣝࢻࣥCynara cardunculus ࢆရ✀ᨵⰋࡋࡓࡶࡢ⪃࠼ࡽࢀ࡚࠸ࡿC,Dࠋ㛤ⰼ┤๓ ࡢⷣࡸⱝⴥࢆ㔝⳯ࡋ࡚㣗ࡿࠊⴥࢆࣁ࣮ࣈࢸ࣮ࡋ࡚㣧⏝ࠊษࡾⰼࡋ࡚㚷㈹ࡉࢀ ࡿ࡞ୡ⏺୰࡛ぶࡋࡲࢀ࡚࠸ࡿEࠋ࣮ࢸࢳ࣮ࣙࢡࡢṔྐࡣྂࡃࠊ⣖ඖ๓ࡢࢠࣜࢩ࣭ ࣮࣐ࣟ௦ࡽ㔝⳯ࡋ࡚㣗ࡉࢀࠊᆅ୰ᾏᆅ᪉ࡢẸ㛫⒪ἲ࡛ࡣ⏕ࡸⴥࡢỒࢆ࣡ࣥΰࡐ ࡚⫢⮚ࡢẸ㛫⸆ࡋ࡚⏝ࡉࢀDࠊ࣮ࣚࣟࢵࣃ࡛ࡣ࣮ࣚࢢࣝࢺΰࡐ࡚⨾ᐜࣃࢵࢡࡋ࡚ ⏝࠸ࡿ࡞Fᵝࠎ࡞ሙ㠃࡛㔜⏝ࡉࢀ࡚࠾ࡾࠊ⌧௦࠾࠸࡚ࡶ⬡⫫ࡸࣝࢥ࣮ࣝࡢ௦ㅰࢆຓ ࡅࡿ㣗ᮦࡋ࡚ᩱ⌮⏝࠸ࡽࢀࡿࠋ⸆⌮స⏝ࡋ࡚ⴥࡸⰼᡸࠊ⥲ⱖ∦ᒀࠊᙉኊࠊ⫹Ồ ศἪಁ㐍ࠊ⾑୰ࢥࣞࢫࢸ࣮ࣟࣝపୗࠊ⫹ಖㆤ࡞ࡢస⏝ࡀ▱ࡽࢀࠊື⬦◳ࡸ㯤⑇ࠊᾘ Ⰻࠊ㣗ḧ⏝࠸ࡽࢀ࡚࠸ࡿG,Hࠋ᪥ᮏࡣỤᡞ௦Ώ᮶ࡋࡓࡀᅜෆ࡛ࡣ୕ᾆ༙ᓥ ᡣ⥲༙ᓥ࡛ࢃࡎ᱂ᇵࡉࢀ࡚࠸ࡿࡢࡳ࡛࠶ࡿCࠋ࣮ࢸࢳ࣮ࣙࢡࡣࠊ⏕⌮άᛶᡂศ ࡋ࡚ࠊࢩࢼࣜࣥࠊࣇࣛ࣎ࣀࢻࠊ࣏ࣜࣇ࢙ࣀ࣮ࣝࠊ࢝ࣇ࢙㓟ࠊࢡࣟࣟࢤࣥ㓟ࠊ࢝ࣟࢸࣥࠊ ࣅࢱ࣑ࣥC ࡸࢭࢫ࢟ࢸࣝ࣌ࣥࣛࢡࢺࣥࡢ㸯ࡘ࡛࠶ࡿࢩࢼࣟࣆࢡࣜࣥࢆྵࡴC,Iࠋᡃࠎࡣࢤࣝ ศ㓄࣒࢝ࣛࢡ࣐ࣟࢺࢢࣛࣇ࣮ࡼࡗ࡚࣮ࢸࢳ࣮ࣙࢡࡢࢩࢼࣟࣆࢡࣜࣥࢆศ㞳ࡋࠊᙉ ࠸NF-κB ㌿άᛶ㜼ᐖస⏝ࡀ᭷ࡿࡇࢆぢฟࡋࡓࠋ
࣮ࢸࢳ࣮ࣙࢡࡣࣝࢥ࣮ࣝࡸD-࢞ࣛࢡࢺࢧ࣑ࣥࡼࡿ⫢ᶵ⬟పୗࢆ㜼ᐖࡍࡿస ⏝ࡀᩘከࡃሗ࿌ࡉࢀ࡚࠾ࡾࠊከᩘࡢࢧࣉ࣓ࣜࣥࢺࡶ㈍ࡉࢀ࡚࠸ࡿ28,29ࠋᡃࠎࡀ࣮ࢸ ࢳ࣮ࣙࢡᢳฟ≀ࢆ⏝࠸࡚࢞ࣛࢡࢺࢧ࣑ࣥࢆ⏝࠸ࡓ⫢ᶵ⬟ホ౯ヨ㦂ࢆᐇࡋࡓࡇࢁࠊሗ࿌ ࡉࢀ࡚࠸ࡿ࠾ࡾࠊD-࢞ࣛࢡࢺࢧ࣑ࣥࡼࡗ࡚ୖ᪼ࡋࡓ⾑୰ࡢ GPT ್ࡀ࣮ࢸࢳ࣮ࣙࢡ ᢳฟ≀ࡢ⤒ཱྀᢞࡼࡾୖ᪼ࡀᢚไࡉࢀࡓࠋࡉࡽࠊ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࡣ↓Ỉ࢚ ࢱࣀ࣮ࣝࡢᢞࡸỈᾐᣊ᮰ࢫࢺࣞࢫࡼࡾㄏⓎࡉࢀࡓ⫶⢓⭷യᐖࡢᢚไࡸ⫶⢓ᾮࡢቑຍࠊ ⫶⢓ᾮྵ㔞ῶᑡࢆ㜵ࡄࡇࡶ☜ㄆࡉࢀࡓࠋࡇࡢຠᯝࡣࠊ࣮ࢸࢳ࣮ࣙࢡࡢᡂศ࡛࠶ࡿࢩ ࢼࣟࣆࢡࣜࣥࡼࡿࡶࡢሗ࿌ࡉࢀ࡚࠸ࡿ30ࠋࡋࡋࠊࡇࢀࡲ࡛㈍ࡉࢀ࡚࠸ࡿ࣮ࢸ ࢳ࣮ࣙࢡࢆཎᩱࡍࡿࢧࣉ࣓ࣜࣥࢺࡣࠊࢩࢼࣜࣥࠊࢡࣟࣟࢤࣥ㓟࡞ࡀᡂศࡋ ࡚ྵࡲࢀ࡚࠸ࡿࡀࢩࢼࣟࣆࢡࣜࣥࡣྵࡲࢀ࡚࠸࡞ࡗࡓࠋࢩࢼࣜࣥࡸࢡࣟࣟࢤࣥ㓟ࡣ NF-κB 㜼ᐖάᛶࡣㄆࡵࡽࢀ࡞ࡗࡓⅭࠊ᪤Ꮡ〇ရ࠾࠸࡚ྠᵝࡢຠᯝࡣᮇᚅ࡛ࡁ࡞࠸ᛮ ࢃࢀࡿࠋࢩࢼࣟࣆࢡࣜࣥࡣ࣮ࢸࢳ࣮ࣙࢡ௨እࡶ࢟ࢲࢳ࢟ࢶࢿࢨ࣑31ࡸᮌ㤶32࡞ ᵝࠎ࡞࢟ࢡ⛉ࡢ᳜≀ࡶྵࡲࢀࡿሗ࿌ࡉࢀ࡚࠸ࡿࡀࠊ᳜≀࠾ࡅࡿࢩࢼࣟࣆࢡࣜࣥྵ᭷ 㔞ࢆẚ㍑ࡋࡓሗ࿌ࡣࡉࢀ࡚࠸࡞࠸ࠋ࣮ࢸࢳ࣮ࣙࢡࡣࠊୡ⏺୰࡛㣗ࡉࢀࡿ㔝⳯ࡢࡦࡘ ࡛࠶ࡿࡀࠊ୍⯡ὶ㏻ࡍࡿⷣ≧ࡢⰼᡸ㒊ศࡣࢩࢼࣟࣆࢡࣜࣥࡣࢇྵࡲࢀ࡚࠾ࡽࡎࠊ ࡑࡢࢇࡣⴥࡢ୰Ꮡᅾࡋ࡚࠸ࡿࠋࡍ࡞ࢃࡕࠊᅇNF-κB 㜼ᐖάᛶࢆぢฟࡋࡓࢩࢼࣟ ࣆࢡࣜࣥࡣࡇࢀࡲ࡛༑ศ࡞◊✲ࡀ࡞ࡉࢀ࡚࠸࡞࠸ࡔࡅ࡛࡞ࡃࠊࡑࡢ⸆⌮άᛶࡶ᭷ຠ⏝ ࡛ࡁ࡚࠸࡞ࡗࡓྍ⬟ᛶࡀ㧗࠸ࠋ ᮏ◊✲࡛ࡣࢩࢼࣟࣆࢡࣜࣥࡢNF-κB ㌿άᛶ㜼ᐖస⏝ࡑࡢస⏝ᶵᗎࠊࡉࡽ࣐࢘ࢫ ࠾࠸࡚ࡢ⤒ཱྀᢞࡼࡿ⓶ග⪁㜵Ṇࡢྍ⬟ᛶࡘ࠸᳨࡚ウࡋࡓࠋ
ྡ⛠ ྵ䜎䜜䜛᳜≀ 䜰䞁䝖䝅䜰䝙䞁 䝤䝗䜴䞉㯮⡿䞉䝤䝹䞊䝧䝸䞊 䝦䝇䝨䝸䝆䞁 䜏䛛䜣䞉䛿䛳䛥䛟 䜹䝔䜻䞁 Ⲕ䞉䝽䜲䞁 䜿䝹䝉䝏䞁 ⋢䛽䛞䞉᯿ᶲ㢮䞉䝋䝞 䜲䝋䝣䝷䝪䞁 ㇋䛺䛹 䝸䜾䝘䞁 䝂䝬 䜽䝻䝻䝀䞁㓟 䝁䞊䝠䞊䞉䛤䜌䛖 䜽䝹䜽䝭䞁 䜴䝁䞁䛺䛹 䝃䝫䝙䞁 䝃䝫䝙䞁 ㇋䞉㧗㯇䝙䞁䝆䞁 䝇䝹䝣䜷䝷䝣䜯䞁 䝤䝻䝑䝁䝸䞊䞉䜻䝱䝧䝒 䜰䝸䝹䜲䝋䝏䜰䝅䝛䞊䝖 䝽䝃䝡 䝅䝇䝔䜲䞁䝇䝹䝩䜻䝅䝗 䝙䞁䝙䜽䞉⋢䛽䛞䞉䜻䝱䝧䝒 β㻙䜹䝻䝔䞁 䝙䞁䝆䞁䞉䛛䜌䛱䜓 α㻙䜹䝻䝔䞁 䝙䞁䝆䞁䞉䛛䜌䛱䜓䞉䝖䜴䝰䝻䝁䝅 䝸䝁䝢䞁 䝖䝬䝖䞉䝇䜲䜹䞉ᰠ 䝹䝔䜲䞁 䝖䜴䝰䝻䝁䝅䞉䝩䜴䝺䞁䝋䜴䞉䝤䝻䝑䝁䝸䞊 䝊䜰䜻䝃䞁䝏䞁 䝖䜴䝰䝻䝁䝅䞉䝩䜴䝺䞁䝋䜴 β㻙䜽䝸䝥䝖䜻䝃䞁䝏䞁 䜏䛛䜣䞉䜸䝺䞁䝆 䜸䜲䝀䝜䞊䝹 䜽䝻䞊䝤 䝸䝰䝛䞁 ᯿ᶲ㢮 䝯䞁䝖䞊䝹 䝝䝑䜹 䝏䝰䞊䝹 䝍䜲䝮䞉䜸䝺䜺䝜 䜲䜸䜴ྜ≀ 䜹䝻䝔䞁㢮 䝔䝹䝨䞁㢮 䝣䝷䝪䝜䜲䝗 䝣䜵䝜䞊䝹㢮 䝫䝸䝣䜵䝜䞊䝹 ✀㢮 ⾲ ⾲⾲ ⾲1 ௦⾲ⓗ࡞௦⾲ⓗ࡞௦⾲ⓗ࡞௦⾲ⓗ࡞Phytochemicals ⾲ ⾲⾲ ⾲2 ⮬↛⪁ග⪁ࡢẚ㍑⮬↛⪁ග⪁ࡢẚ㍑⮬↛⪁ග⪁ࡢẚ㍑⮬↛⪁ග⪁ࡢẚ㍑ ග⪁⓶ ᮏ᮶䛾⪁⓶ ⾲⓶ᒙ ཌ䛔 ⷧ䛔 䜿䝷䝏䝜䝃䜲䝖 ከᵝ䛺⣽⬊ ᆒ୍䛺⣽⬊ ᴟᛶ䛾ᾘኻ ᴟᛶ䛾ಖᏑ 䛧䜀䛧䜀⫧ ㏻ᖖⴎ⦰ 䝯䝷䝜䝃䜲䝖 ⣽⬊ᩘቑຍ ⣽⬊ᩘῶᑡ 䝯䝷䝜䝋䞊䝮⏘⏕ቑຍ 䝯䝷䝜䝋䞊䝮⏘⏕ ከᵝ䛺⣽⬊ ᆒ୍䛺⣽⬊ ங㢌┿⓶ ᡥᖹ ᡥᖹ ⥙≧┿⓶ ⫧ཌᛶᙎᛶⴎ⦰ ⷧ䛟䛺䜛 ᙎᛶ⧄⥔ ᴟ䜑䛶ቑຍ 䜋䛸䜣䛹ṇᖖ ↓ᐃᙧ䛾ሢ䜢ᙧᡂ 䝁䝷䞊䝀䞁 ⥺⥔᮰ཬ䜃⥺⥔䛿ⴭྡ䛻ῶᑡ ⥺⥔᮰䛿ኴ䛟↓㉮ྥ 䜾䝸䝁䝃䝭䝜䜾䝸䜹䞁 ⴭྡ䛻ቑຍ ㍍ᚤῶᑡ ⾲ ⓶ ┿ ⓶
ᅗ ᅗ ᅗ ᅗ1 NF-κκκκB ࡢάᛶࡼࡿ⓶ࡢᙳ㡪ࡢάᛶࡼࡿ⓶ࡢᙳ㡪ࡢάᛶࡼࡿ⓶ࡢᙳ㡪ࡢάᛶࡼࡿ⓶ࡢᙳ㡪 ᅗ ᅗ ᅗ ᅗ2 ྛ✀᳜≀࢚࢟ࢫࡼࡿྛ✀᳜≀࢚࢟ࢫࡼࡿྛ✀᳜≀࢚࢟ࢫࡼࡿྛ✀᳜≀࢚࢟ࢫࡼࡿNF-κκκκB ౫Ꮡࡍࡿ㌿άᛶ㜼ᐖస⏝౫Ꮡࡍࡿ㌿άᛶ㜼ᐖస⏝౫Ꮡࡍࡿ㌿άᛶ㜼ᐖస⏝౫Ꮡࡍࡿ㌿άᛶ㜼ᐖస⏝ ྛ✀᳜≀࢚࢟ࢫࡣ10 ಸ㔞ࡢ 30㸣࢚ࢱࣀ࣮ࣝࢆຍ࠼࡚ᾐₕࡋࠊࢁ㐣ࡋࡓࠋ4κBwt-Luc ࣉࣛࢫ࣑ࢻ࡛ࢺࣛ ࣥࢫࣇ࢙ࢡࢩࣙࣥࡉࡏࡓ293 ⣽⬊ࡣࠊ⤊⃰ᗘࡋ࡚ 1%ࡢ᳜≀࢚࢟ࢫࢆຍ࠼ࡓ 2 㛫ᚋ 1 ng/ml ࡢ TNF-α ࡛ฎ⌮ࡋࡓࠋࡇࢀࡽࡢヨ㦂ࡣᑡ࡞ࡃࡶ3 ᅇ⧞ࡾ㏉ࡉࢀࡓࠋ
ヨ㦂ᮦ ヨ㦂ᮦ ヨ㦂ᮦ ヨ㦂ᮦᩱヨ㦂᪉ἲᩱヨ㦂᪉ἲᩱヨ㦂᪉ἲᩱヨ㦂᪉ἲ 1. ᮦᩱヨ⸆ᮦᩱヨ⸆ᮦᩱヨ⸆ᮦᩱヨ⸆ ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࡣࠊ Cynara scolymus L.ࡢⴥ 15 ಸ㔞ࡢ 45%࢚ࢱࣀ࣮ࣝࢆຍ࠼ ࡚ᾐₕࡋࠊࢁ㐣ᚋࠊࢁᾮࢆ⃰⦰ࠊࡑࡢᚋࠊࢩࢼࣟࣆࢡࣜࣥࡀ1%࡞ࡿࡼ࠺ࠊࢹ࢟ࢫࢺࣜ ࡛ࣥㄪ〇ࡋࠊ⤖⇱ࡋࡓࠋࢩࢼࣟࣆࢡࣜࣥࣃ࢘ࢲ࣮㸦CP㸧ࡣ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ ࡽࢤࣝศ㞳࣒࢝ࣛࢆ⏝࠸࡚⢭〇ࡋࠊࢩࢼࣟࣆࢡࣜࣥࡀ0.65%࡞ࡿࡼ࠺ࢹ࢟ࢫࢺ࡛ࣜࣥㄪ 〇ࡋࡓࠋࢩࢼࣟࣆࢡࣜࣥࡣࠊᕷ㈍ࡢᶆရࢆbionaturis㸦Sevilleࠊࢫ࣌ࣥ㸧ࡽ㉎ධࡋࡓࠋ TNF-αࡣග⣧⸆㸦Osaka, Japan㸧ࡽ㉎ධࡋࡓࠋNF-κB ౫Ꮡⓗ࣍ࢱࣝࣝࢩࣇ࢙࣮ࣛࢮࢆ Ⓨ⌧ࡍࡿpGL3-4kBwt-luc ࣉࣛࢫ࣑ࢻࡣ pGL3-ࣉ࣮ࣟࣔࢱ࣮࣭࣋ࢡࢱ࣮㸦Promega, Madison, W㸧 HIV-1 ࢚ࣥࣁࣥࢧ࣮⏤᮶ࡢ NF-κB ⤖ྜ㓄ิ㸦GGGACTTTCC㸧ࢆ 4 ࢥࣆ࣮ᤄධࡍࡿ ࡇࡼࡗ࡚స〇ࡉࢀࡓ17,33-35ࠋNF-κB ࡢࢧࣈࣘࢽࢵࢺ࡛࠶ࡿ p65 ࢆⓎ⌧ࡍࡿࣉࣛࢫ࣑ࢻ 㸦pCMV-p65㸧ࡣࠊᡭሯࡽࡢሗ࿌ࡢ㏻ࡾస〇ࡋࡓ35ࠋ࣏࣮ࣜࢱ࣮࣭ࣉࣛࢫ࣑ࢻࡣࠊྡྂᒇᕷ ❧Ꮫࠊᒸᮏᩍᤵࡼࡾᥦ౪㡬࠸ࡓࠋ
2. ⣽⬊ᇵ㣴⣽⬊ᇵ㣴⣽⬊ᇵ㣴⣽⬊ᇵ㣴
293 ⣽⬊㸦ࣄࢺ⭈⏤᮶ୖ⓶⣔⣽⬊㸧㸦Riken, Tsukuba, Japan㸧ࡣ 10%FBSࠊ1 mM ࡢࢢࣝࢱ ࣑ࣥ㓟ሷࠊ100 units/ml ࡢ࣌ࢽࢩࣜࣥ 100 mg/ml ࡢࢫࢺࣞࣉࢺ࣐ࢩࣥῧຍࡢ DMEM ࢆ ⏝࠸ࠊ37Υ࡛ᇵ㣴ࡋࡓࠋ⣽⬊ẘᛶࡣࠊMTT ἲࡼࡗ࡚ホ౯ࡋࡓࠋࣄࢺṇᖖ⾲⓶࣓ࣛࣀࢧ ࢺ㸦TOYOBO, Tokyo, Japan㸧ࡣࠊ37Υ࡛ 1㸣࢘ࢩ⫾ඣ⾑Ύࠊ100 units/ml ࣌ࢽࢩࣜࣥࠊ100 μg
/ ml ࡢࢫࢺࣞࣉࢺ࣐ࢩࣥῧຍࡢ࣓ࣛࢽࣥ⣽⬊ᇶᮏᇵᆅ㸦TOYOBO㸧࡛ᇵ㣴ࡋࡓࠋࣄࢺṇ ᖖ⾲⓶ゅ⣽⬊㸦KURABO, Osaka, Japan㸧ࡣࠊࢣࣛࢳࣀࢧࢺᇶᮏᇵᆅ㸦KURABO㸧ࢆ⏝ ࠸ࠊ37Υ࡛ᇵ㣴ࡋࡓࠋࣄࢺṇᖖ⥺⥔ⱆ⣽⬊㸦KURABO㸧ࡣ 5㸣࢘ࢩ⫾ඣ⾑Ύࠊ100 units/ml ࣌ࢽࢩࣜࣥࠊ100 μg/ml ࡢࢫࢺࣞࣉࢺ࣐ࢩࣥῧຍࡢ DMEM ࢆ⏝࠸ࠊ37Υ࡛ᇵ㣴ࡋࡓࠋ
3. 㑇ఏᏊᑟධࣝࢩࣇ࢙࣮ࣛࢮࢵࢭ㑇ఏᏊᑟධࣝࢩࣇ࢙࣮ࣛࢮࢵࢭ㑇ఏᏊᑟධࣝࢩࣇ࢙࣮ࣛࢮࢵࢭ㑇ఏᏊᑟධࣝࢩࣇ࢙࣮ࣛࢮࢵࢭ
⣽⬊Fugene-6 ࢺࣛࣥࢫࣇ࢙ࢡࢩࣙࣥヨ⸆㸦Roche, Basal, Switzerland㸧ࢆ⏝࠸࡚ࣉࣛࢫ ࣑ࢻࢆ㑇ఏᏊᑟධࡋࡓࠋ12well ࣉ࣮ࣞࢺ࡛ᇵ㣴ࡉࢀࡓ⣽⬊ࠊ1.5 ml ࡢ Fugene-6 ࠾ࡼࡧ 0.5 μg ࡢࣉࣛࢫ࣑ࢻ DNA ࢆΰྜࡋ࡚ᑟධࡋࡓ17,34-36ࠋ㑇ఏᏊᑟධࡢ24 㛫ᚋࠊヨ㦂ヨᩱࢆῧ ຍࡋࡓࠋࡑࡢ2 㛫ᚋ TNF-αࢆῧຍࡋ࡚ NF-κB ࢆάᛶࡉࡏࠊࡉࡽ 24 㛫ᇵ㣴ࡋࡓ ࡢࡕᅇࡋࡓࠋᅇࡋࡓ⣽⬊୰ࡢࣝࢩࣇ࢙࣮ࣛࢮάᛶࡣࠊࣝࢩࣇ࢙࣮ࣛࢮࢵࢭࢩࢫࢸ ࣒㸦Promega㸧࡛ ᐃࡋࡓࠋ ᐃࡣࠊ࣑ࣝࣀ࣓࣮ࢱ࣮TD-20/20㸦Promega㸧ࢆ⏝࠸ࡓࠋ㑇 ఏᏊᑟධຠ⋡ࡣࠊෆ㒊ᶆ‽ࡋ࡚pRL-TK ࣉࣛࢫ࣑ࢻࢆ⏝࠸ࠊ࣍ࢱࣝࣝࢩࣇ࢙࣮ࣛࢮάᛶ ್ࢆ࣑࢘ࢩࢱࢣࣝࢩࣇ࢙࣮ࣛࢮάᛶ್ࡼࡗ࡚㝖ࡍࡿࡇ࡛⿵ṇࡋࡓࠋヨ㦂ࡣࢺࣜࣉࣜ ࢣ࣮ࢺ࡛⾜ࢃࢀࠊ⊂❧ࡋࡓ⣔࡛3 ᅇ௨ୖᐇࡋࡓࠋ 4. bFGF MMP-1 ࡢᐃ㔞ࡢᐃ㔞ࡢᐃ㔞ࡢᐃ㔞
ᕷ㈍ࡢEIA ࢟ࢵࢺ㸦bFGF㸸Cytimmune, College Park, MD; MMP-1㸸Amersham Biosciences, Uppsala, Sweden㸧ࢆ⏝࠸࡚ࠊᣦᐃࡉࢀࡓࣉࣟࢺࢥ࣮ࣝᚑࡗ࡚⾜ࡗࡓࠋヨ㦂ࡣࢺࣜࣉࣜࢣ ࣮ࢺ࡛⾜ࢃࢀࠊ⊂❧ࡋࡓ⣔࡛2 ᅇ௨ୖᐇࡋࡓࠋ
5. ࣓ࣛࣀࢧࢺቑṪ ᐃ࣓ࣛࣀࢧࢺቑṪ ᐃ࣓ࣛࣀࢧࢺቑṪ ᐃ࣓ࣛࣀࢧࢺቑṪ ᐃ
ࢣࣛࢳࣀࢧࢺࡢᇵ㣴ୖΎࡼࡿ࣓ࣛࣀࢧࢺ⏘⏕ಁ㐍άᛶࢆホ౯ࡍࡿࡓࡵࢣࣛࢳࣀ ࢧࢺpCMV-p65Vedtor ࢆᑟධࡋࠊ24 㛫ᇵ㣴ᚋᇵᆅࢆࡋࡓࠋࡑࡢᚋࠊヨ㦂ヨᩱࢆ ῧຍࡋࠊ48 㛫ᇵ㣴ᚋୖΎࢆ᥇ྲྀࡋࡓࠋ࣓ࣛࣀࢧࢺࡣ⣽⬊ಁ㐍ᮍῧຍࡢ Medium154S 㸦KURABO㸧ࢣࣛࢳࣀࢧࢺᇵ㣴ୖΎࢆ 1㸸4 ΰྜࡋࡓᇵᆅࡋࠊࡉࡽ 72 㛫 ᇵ㣴ᚋࠊ⣽⬊ࢆᅇࡋࠊ࣓ࣛࣀࢧࢺᩘࢆ⣽⬊ᩘ ᐃヨ⸆WST-1㸦Roche, Basal, Switzerland㸧 ࢆ⏝࠸࡚ ᐃࡋࡓࠋ
6. චᰁⰍࡼࡿචᰁⰍࡼࡿචᰁⰍࡼࡿචᰁⰍࡼࡿp65 ࡢᒁᅾࡢほᐹࡢᒁᅾࡢほᐹࡢᒁᅾࡢほᐹࡢᒁᅾࡢほᐹ
NF-κB ࡢ⣽⬊ෆᒁᅾࢆㄪࡿࡓࡵࠊ293 ⣽⬊ヨ㦂ヨᩱࢆῧຍᚋ 3 㛫ᇵ㣴ࡋࡓࠋ TNF-α㸦1 ng/ml㸧ῧຍࡼࡾ NF-κB ࢆάᛶࡋࠊ25 ศᚋ⣽⬊ࢆᅛᐃࡋࡓࠋࡲࡓࠊࢥࣥ ࢺ࣮ࣟࣝ⩌ࡋ࡚TNF-αᮍῧຍ⩌ࢆタࡅࡓࠋ
୍ḟᢠయࡋ࡚࢘ࢧࢠ࣭࣏ࣜࢡ࣮ࣟࢼࣝp65 ᢠయ㸦Santa Cruz Biotechnology, Inc., Santa Cruz, CA㸧ࠊḟᢠయࡋ࡚ࣇࣝ࢜ࣞࢭࣥࢯࢳ࢜ࢩࢿ࣮ࢺ࡛ᶆ㆑ࡉࢀࡓࣖࢠᢠ࢘ࢧࢠ IgG㸦Cappel, Irvine, CA㸧ࢆ⏝࠸࡚චᰁⰍࡋࠊp65 ࡢᒁᅾࢆㄪࡓ37ࠋ
7. UVB ↷ᑕ⓶ࣔࢹ࣐ࣝ࢘ࢫ↷ᑕ⓶ࣔࢹ࣐ࣝ࢘ࢫ↷ᑕ⓶ࣔࢹ࣐ࣝ࢘ࢫ↷ᑕ⓶ࣔࢹ࣐ࣝ࢘ࢫ
5 㐌㱋ࡢ㞝ᛶ DBA/2 ࣐࢘ࢫ 29 ༉ࢆ⏝ࡋࡓࠋ࣐࢘ࢫࡣ⣸እ⥺↷ᑕ⩌ࠊ⣸እ⥺↷ᑕ + ࢩ ࢼࣟࣆࢡࣜࣥࣃ࢘ࢲ࣮㸦CP㸧462.2 mg/kg㸦3.0 mg/kg ࢩࢼࣟࣆࢡࣜࣥ┦ᙜ㸧ᢞ⩌ࠊ⣸ እ⥺↷ᑕ + CP 92.4 mg/kg㸦0.6 mg/kg ࢩࢼࣟࣆࢡࣜࣥ┦ᙜ㸧ᢞ⩌ࠊࢥࣥࢺ࣮ࣟࣝ⩌
ศࡅࡽࢀࡓࠋ࣐࢘ࢫࡣẖ᪥ࠊCP ࡲࡓࡣࢥࣥࢺ࣮ࣟࣝࡋ࡚ヨ㦂ヨᩱస〇⏝ࡋࡓࢹ࢟ ࢫࢺࣜࣥࢆࢰࣥࢹࢆ⏝࠸࡚⤒ཱྀᢞࡋࡓࠋ⣸እ⥺↷ᑕࡣࠊ12 ᪥㛫࡛ 6 ᅇࠊ280㹼320 nm Ἴ 㛗ࡢ⣸እ⥺ࢆFL20 SE ࢧࣥࣛࣥࣉ㸦TOSHIBA, Tokyo, Japan㸧ࢆ⏝࠸࡚ 180 mJ/cm2ࡢ⏝㔞࡛ ࣐࢘ࢫ↷ᑕࡋࡓࠋヨ㦂ᮇ㛫୰ࡣ⤒ཱྀᢞࢆ⾜ࡗࡓᚋࠊ㐌3 ᅇࠊ⣸እ⥺↷ᑕࢆ⾜ࡗࡓࠋ12 ᪥ᚋࠊࡍ࡚ࡢ࣐࢘ࢫࡽ⪥ࢆ᥇ྲྀࡋ∦ഃࡣࣃࣛࣇࣥໟᇙᶆᮏࠊࡶ࠺୍᪉ࡣ-80Υ ಖᏑࡋࡓᚋࠊ࣓ࣛࣀࢧࢺᩘࡢィ ⏝࠸ࡓࠋࣃࣛࣇࣥໟᇙࡋࡓ⪥ࢆ⏝࠸࡚⾲⓶ࡢཌ ࡉࢆ ᐃࡍࡿࡓࡵࠊ࣐࣊ࢺ࢟ࢩ̺࢚ࣜࣥ࢜ࢪࣥᰁⰍ㸦HE ᰁⰍ㸧ࢆ⾜࠸ࠊගᏛ㢧ᚤ㙾ୗ࡛ ᙳࡋࠊ⏬ീゎᯒࢯࣇࢺ㸦WINROOF㸹Mitani, Fukui, Japan㸧ࢆ⏝ࡋ࡚ᩘ್ࡋࡓࠋ
ࡲࡓࠊචᰁⰍࡼࡗ࡚ቑṪ⣽⬊᰾ᢠཎ㸦PCNA㸧ࡢⓎ⌧ࢆホ౯ࡋࡓࠋࣃࣛࣇࣥໟᇙ ᶆᮏࢆ3 μm ษࡾศࡅࠊ⬺ࣃࣛࣇࣥࢆ⾜ࡗࡓᚋࠊDako Target Retrieval Solution㸦pH 4.0; Dako, Glostrup, Denmark㸧࡛ 121Υࠊ5 ศ㛫ᛂࡋᢠཎࢆ㟢ฟࡉࡏࡓࠋࣈࣟࢵ࢟ࣥࢢ⁐ᾮ㸦㸣 ࢘ࢩ⾑Ύࣝࣈ࣑ࣥࠊ㸣࢘ࢩ⫾ඣ⾑Ύ㸧ࡢධࡗࡓPBS ࡛ᛂᚋࠊᢠ PCNA ึᮇᢠయ㸦 abcam, Cambridge, U.K.㸧࡛ Υࠊ ศ㛫ᛂࡉࡏࡓࠋࡉࡽࠊAlexa Fluor 594 goat anti-rabbit IgG 㸦H+L㸧㸦1:200, Molecular Probes, Eugene, OR㸧࡛ࠊᐊ ࡛ ศ㛫ᛂࡉࡏࡓࠋࡲࡓࠊ ᰾ࢆ࣊࢟ࢫࢺ ⁐ᾮ㸦 Dojindo Laboratories, Kumamoto, Japan㸧࡚ᰁⰍࡋࡓࠋ චᰁⰍᚋࠊ⺯ග㢧ᚤ㙾㸦㸧㸦Olympus, Tokyo, Japan㸧ࢆ⏝ࡋ࡚ほᐹࡋࡓࠋ
⓶⤌⧊୰ࡢ࣓ࣛࣀࢧࢺᩘࡢィ ࡣࠊHiramoto ࡽ38ࡢ᪉ἲࡼࡗ࡚⾜ࡗࡓࠋ᥇ྲྀࡋࡓ ⪥ࡽ㌾㦵ࢆྲྀࡾ㝖ࡁࠊ37Υ⎔ቃୗ࡛ 2N NaBr ⁐ᾮ㸰㛫ᾐₕࡋࡓࠋࡑࡢᚋࠊ⓶⤌ ⧊ࡽ⾲⓶࠾ࡼࡧᇶᗏᒙࢆ㞳ࡋࠊDOPA ᰁⰍࢆࡋࡓࠋ㢧ᚤ㙾ୗ࡛ 5 mm2ࡢDOPA 㝧ᛶ࣓ ࣛࣀࢧࢺᩘࢆ࢝࢘ࣥࢺࡋࠊ1 mm2࠶ࡓࡾࡢ⣽⬊ᩘ࡛♧ࡋࡓࠋ
ࡇࢀࡽࡢື≀ᐇ㦂ࡣࠊ⌮ㄪᰝጤဨࡢᢎㄆࢆᚓࡓᚋࠊᐇࡉࢀࡓࠋ 8. ⤫ィศᯒ⤫ィศᯒ⤫ィศᯒ⤫ィศᯒ ࢹ࣮ࢱࡣࠊᑡ࡞ࡃࡶ3 ᅇࡢ⊂❧ࡋࡓᐇ㦂ࡽ㞟ࡵࡽࢀࡓࠋື≀ᐇ㦂ࡣࠊྛ⩌ࡘࡁᑡ ࡞ࡃࡶ7 ༉ࡢື≀࡛ᐇ⾜ࡉࢀࡓࠋࢹ࣮ࢱࡣࠊᖹᆒ +/- S.D.ࡋ࡚♧ࡋࠊ⤫ィⓗ᭷ពᛶࡣࠊ ANOVAࡼࡗ࡚ㄪࡽࢀࡓࠋ p㸺0.05 ࡞ࡽࡤࠊ⤫ィⓗ᭷ពᕪ࠶ࡾࡋࡓࠋ ⤖ᯝ ⤖ᯝ ⤖ᯝ ⤖ᯝ 1. NF-κκκκB ㌿άᛶ㜼ᐖస⏝㌿άᛶ㜼ᐖస⏝㌿άᛶ㜼ᐖస⏝㌿άᛶ㜼ᐖస⏝ ᅗ3A ࡛♧ࡍࡼ࠺ࠊࣄࢺ⭈⏤᮶ୖ⓶⣔⣽⬊ 293 ⣽⬊ࡣ TNF-α่⃭ࡼࡾ NF-κB ࡢ㌿ άᛶࡣቑຍࡋࡓࠋࡋࡋTNF-αῧຍ๓࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢆῧຍࡋࡓࡁࠊ⃰ᗘ ౫Ꮡⓗࡑࡢάᛶࢆ㜼ᐖࡋࡓ㸦ᅗ3A㸧ࠋ ࣮ࢸࢳ࣮ࣙࢡྵࡲࢀࡿࢭࢫ࢟ࢸࣝ࣌ࣥࣛࢡࢺࣥࡢ1 ✀࡛࠶ࡿࢩࢼࣟࣆࢡࣜࣥ 㸦ᅗ3B㸧ࡣ NF-κB ࡢ㌿άᛶ㜼ᐖస⏝ࢆ♧ࡋࡓ㸦ᅗ 3C㸧ࠋࡲࡓࠊ⏝࠸ࡓ⃰ᗘ࠾࠸࡚᭷ ព࡞⣽⬊ẘᛶࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢩࢼࣟࣆࢡࣜࣥࡢNF-κB ㌿ άᛶࢆྠ୍ࢢࣛࣇୖ♧ࡋࡓ㸦ᅗ3D㸧ࠋ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ྵࡲࢀࡿࢩࢼࣟࣆࢡ ࣜࣥ⃰ᗘࢆX ㍈ୖ࡛ᥞ࠼ࡓࡁࠊ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢩࢼࣟࣆࢡࣜࣥࡢ NF-κB ά ᛶ㜼ᐖࡣྠ⛬ᗘ࡛࠶ࡗࡓࠋࡇࡢ⤖ᯝࡣࠊNF-κB ࡢ㌿άᛶࢆ㜼ᐖࡍࡿ࣮ࢸࢳ࣮ࣙࢡᢳ
ฟ≀ࡢάᛶᡂศࡀࢩࢼࣟࣆࢡ࡛ࣜࣥ࠶ࡿྍ⬟ᛶࡀᴟࡵ࡚㧗ࡃࠊ┦ⓗࡶࡋࡃࡣ┦ຍⓗస ⏝ࡍࡿࡢ≀㉁ࡀ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࡣྵࡲࢀ࡞࠸ࡇࢆ♧ࡋ࡚࠸ࡿࠋ
B A
ᅗ ᅗ ᅗ ᅗ3 ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢩࢼࣟࣆࢡࣜࣥࡼࡿ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢩࢼࣟࣆࢡࣜࣥࡼࡿ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢩࢼࣟࣆࢡࣜࣥࡼࡿ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢩࢼࣟࣆࢡࣜࣥࡼࡿNF-κκκκB ㌿άᛶ㜼ᐖస⏝㌿άᛶ㜼ᐖస⏝㌿άᛶ㜼ᐖస⏝㌿άᛶ㜼ᐖస⏝ 㸦A㸧ᕥᅗ㸸࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࡢ NF-κB ㌿άᛶࡢຠᯝࠋ4κBwt-Luc ࣉࣛࢫ࣑ࢻ࡛ࢺࣛࣥࢫࣇ ࢙ࢡࢩࣙࣥࡉࡏࡓ293 ⣽⬊ࡣࠊ⤊⃰ᗘࡋ࡚ 140ࠊ280 ࡲࡓࡣ 560 μg/mL ࡢ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢆ ຍ࠼ࡓ2 㛫ᚋ 1 ng/ml ࡢ TNF-α࡛ฎ⌮ࡋࡓࠋࡇࢀࡽࡢヨ㦂ࡣᑡ࡞ࡃࡶ 3 ᅇ⧞ࡾ㏉ࡉࢀࡓࠋྑᅗ㸸 ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࡢ⣽⬊ẘᛶ ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࡼࡿ⣽⬊ẘᛶࡣほᐹࡉࢀ࡞ࡗࡓࠋࢹ࣮ ࢱࡣᖹᆒsS.D.ࠊ᭷ពᕪࡣ ANOVA ࡼࡾ ** p <0.01ࠊn.s. ᭷ពᕪ࡞ࡋ♧ࡋࡓࠋ 㸦B㸧ࢩࢼࣟࣆࢡࣜࣥࡢᏛᵓ㐀 㸦C㸧ᕥᅗ㸸ࢩࢼࣟࣆࢡࣜࣥࡢ NF-κB ㌿άᛶࡢຠᯝࠋ293 ⣽⬊ࡣࢩࢼࣟࣆࢡࣜࣥ㸦2ࠊ4 ࡲࡓࡣ 8 μM㸧 ࡛๓ฎ⌮ࡉࢀࠊ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ྠᵝࡢ᪉ἲ࡛ࣝࢩࣇ࢙࣮ࣛࢮάᛶࢆ ᐃࡋࡓࠋྑᅗ㸸ࢩࢼࣟ ࣆࢡࣜࣥࡢ⣽⬊ẘᛶ ࢩࢼࣟࣆࢡࣜࣥࡼࡿ⣽⬊ẘᛶࡣほᐹࡉࢀ࡞ࡗࡓࠋࢹ࣮ࢱࡣᖹᆒsS.D.ࠊ᭷ព ᕪࡣANOVA ࡼࡾ ** p <0.01ࠊn.s. ᭷ពᕪ࡞ࡋ♧ࡋࡓࠋ 㸦D㸧࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࢩࢼࣟࣆࢡࣜࣥࡢ NF-κB ㌿άᛶ㜼ᐖࡢẚ㍑ C D
2. ࢩࢼࣟࣆࢡࣜࣥࡼࡿࢩࢼࣟࣆࢡࣜࣥࡼࡿࢩࢼࣟࣆࢡࣜࣥࡼࡿࢩࢼࣟࣆࢡࣜࣥࡼࡿNF-κκκκB ࡢస⏝ᶵᗎࡢస⏝ᶵᗎࡢస⏝ᶵᗎࡢస⏝ᶵᗎ ࢩࢼࣟࣆࢡࣜࣥࡼࡿNF-κB ࡢ㌿㜼ᐖ࣓࢝ࢽࢬ࣒ࢆ᫂ࡽࡍࡿࡓࡵࠊᡃࠎࡣࠊ293 ⣽⬊pCMV -p65 ࢆࢺࣛࣥࢫࣇ࢙ࢡࢩࣙࣥࡋࠊp65 㑇ఏᏊࢆ㐣Ⓨ⌧ࡉࡏࡓࠋp65 㑇ఏᏊ ࡢ㐣Ⓨ⌧ࡣࠊI-κB ࡢࣜࣥ㓟ࡸศゎ࡞ࡢ NF-κB ࡢ᰾⛣⾜ࡘ࡞ࡀࡿṇᖖ࡞ࢩࢢࢼࣝఏ 㐩⤒㊰ࢆࣂࣃࢫࡋ࡚ࠊ௵ពࡢ่⃭࡞ࡋNF-κB ᶆⓗ㑇ఏᏊࡢ㌿ᑐࡍࡿᙳ㡪ࢆࡳࡿࡇ ࡀ࡛ࡁࡿࠋᅗ4A ♧ࡍࡼ࠺ࠊࢩࢼࣟࣆࢡࣜࣥࡣ p65 ࡼࡿ㌿άᛶࢆ TNF-α่⃭ ࡼࡿ㌿άᛶྠᵝ⃰ᗘ౫Ꮡⓗ㜼ᐖࡋࡓࠋࡇࢀࡽࡢ▱ぢࡣࠊࢩࢼࣟࣆࢡࣜࣥࡢせ࡞ ࢱ࣮ࢤࢵࢺࡣI-κB ࡢࣜࣥ㓟ศゎࡢୗὶ࡛࠶ࡿࡇࢆ♧ࡋ࡚࠸ࡿࠋࡲࡓࠊ᰾⛣⾜࠾ࡅ ࡿࢩࢼࣟࣆࢡࣜࣥࡢຠᯝࢆNF-κB㸦p65㸧ࡢච⺯ගἲࡼࡾㄪࡓࠋTNF-α่⃭ࡼࡾㄏ ⓎࡉࢀࡓNF-κB ࡢ᰾⛣⾜ࡣࢩࢼࣟࣆࢡࣜࣥࡼࡾ㜼ᐖࡉࢀࡓ㸦ᅗ 4B㸧ࠋ ᅗ ᅗ ᅗ ᅗ4 p65 ࡢ㌿άᛶࡢ㌿άᛶࡢ㌿άᛶࡢࢩࢼࣟࣆࢡࣜࣥࡢ㜼ᐖస⏝㸦ࡢ㌿άᛶࡢࢩࢼࣟࣆࢡࣜࣥࡢ㜼ᐖస⏝㸦ࡢࢩࢼࣟࣆࢡࣜࣥࡢ㜼ᐖస⏝㸦ࡢࢩࢼࣟࣆࢡࣜࣥࡢ㜼ᐖస⏝㸦A㸧㸧㸧㸧NF-κκκκB ࡢ᰾⛣⾜ᢚไ㸦ࡢ᰾⛣⾜ᢚไ㸦ࡢ᰾⛣⾜ᢚไ㸦ࡢ᰾⛣⾜ᢚไ㸦B㸧㸧㸧㸧 㸦A㸧p65 ࢆ㐣Ⓨ⌧ࡉࡏࡓ 293 ⣽⬊ࢩࢼࣟࣆࢡࣜࣥࢆ 2ࠊ4 ࡲࡓࡣ 8 μM ຍ࠼ࡓࠋࢹ࣮ࢱࡣᖹᆒsS.D.ࠊ ᭷ពᕪࡣANOVA ࡼࡾ ** p <0.01 ♧ࡋࡓࠋ 㸦B㸧ࢩࢼࣟࣆࢡࣜࣥ(8 μM)ࡣ TNF-αῧຍ๓ຍ࠼ࡽࢀࡓࠋ A B
3. bFGF ࠾ࡼࡧ࠾ࡼࡧ࠾ࡼࡧ࠾ࡼࡧMMP-1 ⏘⏕࠾ࡅࡿ⏘⏕㜼ᐖస⏝⏘⏕࠾ࡅࡿ⏘⏕㜼ᐖస⏝⏘⏕࠾ࡅࡿ⏘⏕㜼ᐖస⏝⏘⏕࠾ࡅࡿ⏘⏕㜼ᐖస⏝ ⣸እ⥺ㄏⓎࡉࢀࡿ⓶ࡢኚࡣࠊbFGF MMP-1 ࡼࡗ࡚፹ࡉࢀࡿࡇࡀ▱ࡽࢀ ࡚࠸ࡿ8,39ࠋNF-κB ࡢࢧࣈࣘࢽࢵࢺ࡛࠶ࡿ p65 ࢆᑟධࡋࡓࢣࣛࢳࣀࢧࢺ࡛ࡣࠊᖖ NF-κB ࡀάᛶࡋࡓ≧ែ࡞ࡾࠊbFGF MMP-1 ࡢ⏘⏕ࢆㄏᑟࡉࢀࡿࡇࡀሗ࿌ࡉࢀࡓ18,34ࠋᅗ 5A ♧ࡍ࠾ࡾࠊࢩࢼࣟࣆࢡࣜࣥࡣ NF-κB 㸦p65㸧ࡀ㐣Ⓨ⌧ࡋࡓࢣࣛࢳࣀࢧࢺࡢ bFGF ⏘⏕ࢆ᭷ព㜼ᐖࡋࡓࠋྠᵝࠊᅗ5B ࡣࢩࢼࣟࣆࢡࣜࣥࡀ⓶⥺⥔ⱆ⣽⬊ࡽࡢ MMP-1 ⏘⏕ࡶ᭷ព㜼ᐖࡍࡿࡇࢆ♧ࡋ࡚࠸ࡿࠋ ᅗ ᅗ ᅗ ᅗ5 p65 ࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺ࠾ࡅࡿࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺ࠾ࡅࡿࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺ࠾ࡅࡿࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺ࠾ࡅࡿbFGF ⏘⏕㸦⏘⏕㸦⏘⏕㸦⏘⏕㸦A㸧㸧㸧㸧p65 ࢆ㐣Ⓨ⌧ࡉࡏࢆ㐣Ⓨ⌧ࡉࡏࢆ㐣Ⓨ⌧ࡉࡏࢆ㐣Ⓨ⌧ࡉࡏ ࡓ⥺⥔ⱆ⣽⬊࠾ࡅࡿ ࡓ⥺⥔ⱆ⣽⬊࠾ࡅࡿ ࡓ⥺⥔ⱆ⣽⬊࠾ࡅࡿ ࡓ⥺⥔ⱆ⣽⬊࠾ࡅࡿMMP-1 ⏘⏕㸦⏘⏕㸦⏘⏕㸦⏘⏕㸦B㸧㸧㸧ࡢࢩࢼࣟࣆࢡࣜࣥࡢຠᯝ㸧ࡢࢩࢼࣟࣆࢡࣜࣥࡢຠᯝࡢࢩࢼࣟࣆࢡࣜࣥࡢຠᯝࡢࢩࢼࣟࣆࢡࣜࣥࡢຠᯝ ⾲⓶ゅ⣽⬊⥺⥔ⱆ⣽⬊ࡣࢩࢼࣟࣆࢡࣜࣥ4 μM ࢆຍ࠼࡚ヨ㦂ࡋࡓࠋࢹ࣮ࢱࡣᖹᆒsS.D.ࠊ᭷ពᕪࡣ ANOVA ࡼࡾ ** p <0.01ࠊ * p <0.05 ♧ࡋࡓࠋ A B
4. ࣓ࣛࣀࢧࢺቑṪ࠾ࡅࡿ㜼ᐖຠᯝ࣓ࣛࣀࢧࢺቑṪ࠾ࡅࡿ㜼ᐖຠᯝ࣓ࣛࣀࢧࢺቑṪ࠾ࡅࡿ㜼ᐖຠᯝ࣓ࣛࣀࢧࢺቑṪ࠾ࡅࡿ㜼ᐖຠᯝ ⓶࠾࠸࡚࣓ࣛࢽࣥⰍ⣲ࢆసࡿ⾲⓶࣓ࣛࣀࢧࢺࡣࠊ⣸እ⥺ࡸ៏ᛶⓗ࡞⅖➼࡛ቑ ຍࡋࠊࡑࢀࡀ⓶࠾ࡅࡿ࣓ࣛࢽࣥ⏘⏕㔞ࡢቑࡘ࡞ࡀࡿ࠸ࢃࢀ࡚࠸ࡿ8ࠋ࣓ࣛࢽࣥ ⏘⏕㔞ࡢቑຍࡋࡓ⓶࡛ࡣࠊ࣓ࣛࣀࢧࢺᩘࡀቑ࠼࡚࠾ࡾࠊࡑࡢ㐣⛬ࢣࣛࢳࣀࢧࢺෆ ࡛సࡽࢀࡿNF-κB ౫Ꮡᛶ bFGF ࡀ㛵ࡋ࡚࠸ࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ17ࠋNF-κB 㸦p65㸧 ࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺࡢᇵ㣴ୖΎࢆຍ࠼࡚ࠊ࣓ࣛࣀࢧࢺࢆᇵ㣴ࡋࡓࡇࢁࠊ ࣓ࣛࣀࢧࢺࡢ᭷ព࡞ቑຍࡀほᐹࡉࢀࡓࠋࡋࡋࠊ࠶ࡽࡌࡵࢣࣛࢳࣀࢧࢺࢩࢼࣟࣆ ࢡࣜࣥࢆῧຍࡋ࡚ᇵ㣴ࡍࡿ࣓ࣛࣀࢧࢺࡢቑṪࡣ㜼ᐖࡉࢀࡓ㸦ᅗ6AࠊB㸧ࠋ ᅗ ᅗ ᅗ ᅗ p65 ࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺᇵ㣴ୖΎࡢ࣓ࣛࣀࢧࢺቑຍᑐࡍࡿࢩࢼࣟࣆࢡࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺᇵ㣴ୖΎࡢ࣓ࣛࣀࢧࢺቑຍᑐࡍࡿࢩࢼࣟࣆࢡࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺᇵ㣴ୖΎࡢ࣓ࣛࣀࢧࢺቑຍᑐࡍࡿࢩࢼࣟࣆࢡࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺᇵ㣴ୖΎࡢ࣓ࣛࣀࢧࢺቑຍᑐࡍࡿࢩࢼࣟࣆࢡ ࣜࣥࡢຠᯝ ࣜࣥࡢຠᯝ ࣜࣥࡢຠᯝ ࣜࣥࡢຠᯝ 㸦A㸧p65 ࢆ㐣Ⓨ⌧ࡉࡏࡓࢣࣛࢳࣀࢧࢺࢩࢼࣟࣆࢡࣜࣥ 4 μM ࢆຍ࠼࡚ヨ㦂ࡋࡓࠋࢹ࣮ࢱࡣᖹᆒ sS.D.ࠊ᭷ពᕪࡣ ANOVA ࡼࡾ* p <0.05 ♧ࡋࡓࠋ 㸦B㸧ࢣࣛࢳࣀࢧࢺᇵ㣴ୖΎࡢ࣓ࣛࣀࢧࢺࡢస⏝ࠋ B A
5. ࢩࢼࣟࣆࢡࣜࣥࡢ࣐࢘ࢫ⤒ཱྀᢞࡼࡿ⾲⓶⫧ཌ࠾ࡼࡧ࣓ࣛࢽࣥ⏘⏕ࡢᢚไຠᯝࢩࢼࣟࣆࢡࣜࣥࡢ࣐࢘ࢫ⤒ཱྀᢞࡼࡿ⾲⓶⫧ཌ࠾ࡼࡧ࣓ࣛࢽࣥ⏘⏕ࡢᢚไຠᯝࢩࢼࣟࣆࢡࣜࣥࡢ࣐࢘ࢫ⤒ཱྀᢞࡼࡿ⾲⓶⫧ཌ࠾ࡼࡧ࣓ࣛࢽࣥ⏘⏕ࡢᢚไຠᯝࢩࢼࣟࣆࢡࣜࣥࡢ࣐࢘ࢫ⤒ཱྀᢞࡼࡿ⾲⓶⫧ཌ࠾ࡼࡧ࣓ࣛࢽࣥ⏘⏕ࡢᢚไຠᯝ ⣸እ⥺ࡼࡿ⾲⓶ࡢ⫧ཌࡸⰍ⣲ỿ╔ࡀࠊ㐣࡞bFGF ࡼࡿࢣࣛࢳࣀࢧࢺ8,10,11࣓ ࣛࣀࢧࢺࡢ㐣ቑṪࡽ⏕ࡌࡿࡇࡣࡼࡃ▱ࡽࢀ࡚࠸ࡿ39,40ࠋࡲࡓࠊNF-κB ᢚไࡢ⏝ ࡀUVB ↷ᑕࡼࡿ⾲⓶ࡢ⫧ཌࢆᢚไࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ17,18,34ࠋᮏヨ㦂࡛ࡣUVB ↷ᑕࡼࡾㄏⓎࡉࢀࡿ⓶㞀ᐖࡢࢩࢼࣟࣆࢡࣜࣥࣃ࢘ࢲ࣮㸦CP㸧ࡢ⤒ཱྀᢞࡼࡿᙳ㡪 ࢆㄪࡓࠋᅗ7AࠊB ♧ࡍ࠾ࡾࠊࢥࣥࢺ࣮ࣟࣝ⩌ẚ UV ↷ᑕ⩌ࡀ 1.7 ಸࡢ⾲⓶ࡢ⫧ ཌࢆㄏⓎࡋࡓࡢᑐࡋ࡚ࠊ462.2 mg/kg CP㸦3.0 mg/kg ࡢࢩࢼࣟࣆࢡࣜࣥ➼ࡋ࠸㸧ᢞ⩌ ࡛ࡣࠊࢥࣥࢺ࣮ࣟࣝ⩌ྠ⛬ᗘࡲ࡛ᢚไࡋࡓࠋ ྠᵝࠊ92.4 mg/kg CP㸦0.6 mg/kg ࡢࢩࢼ ࣟࣆࢡࣜࣥ➼ࡋ࠸㸧ᢞ⩌࡛ࡣࠊ⾲⓶ࡢ⫧ཌࢆ᭷ពῶᑡࡉࡏࡓࠋࡇࢀࡽࡢ⤖ᯝࡣࠊࢩ ࢼࣟࣆࢡࣜࣥࡀ⃰ᗘ౫Ꮡⓗࢣࣛࢳࣀࢧࢺࡢ㐣ቑṪࢆᢚไࡋࡓࡇࢆ♧ࡋ࡚࠸ࡿࠋᅗ7C ♧ࡋࡓචᰁⰍࡼࡾ᳨ドࡋࡓPCNA 㝧ᛶ⣽⬊ᩘࡣࡍ࡚ࡢ⩌࡛㐪࠸ࢆㄆࡵ࡞ࡗࡓࠋ ࡋࡋࠊUV ↷ᑕ⩌࡛ࡣ᭷᰾⣽⬊ᩘࡀ㢧ⴭቑຍࡋ࡚࠸ࡓࡢᑐࡋࠊCP ᢞ⩌࡛ࡣ᭷᰾⣽ ⬊ࡢቑຍࡣㄆࡵࡽࢀ࡞ࡗࡓࠋ ࡲࡓࠊDOPA 㝧ᛶ࣓ࣛࣀࢧࢺᩘࢆẚ㍑ࡋࡓࡇࢁࠊࢥࣥࢺ࣮ࣟࣝ⩌ẚ UV ↷ᑕ⩌ ࡛ࡣ4.3 ಸࡢ࣓ࣛࣀࢧࢺቑຍࢆㄆࡵࡓࡀࠊ92.4 mg/kg ࡲࡓࡣ 462.2 mg/kg ࡢ CP ᢞ⩌࡛ ࡣ࣓ࣛࣀࢧࢺࡢቑຍࡀ᭷ពᢚไࡉࢀࡓ㸦ᅗ8AࠊB㸧ࠋ
A
ᅗ ᅗ ᅗ ᅗ7 UVB↷ᑕ࡛ㄏⓎࡉࢀࡿ⾲⓶ࡢ⫧ཌᑐࡍࡿࢩࢼࣟࣆࢡࣜࣥࡢᢚไ↷ᑕ࡛ㄏⓎࡉࢀࡿ⾲⓶ࡢ⫧ཌᑐࡍࡿࢩࢼࣟࣆࢡࣜࣥࡢᢚไ↷ᑕ࡛ㄏⓎࡉࢀࡿ⾲⓶ࡢ⫧ཌᑐࡍࡿࢩࢼࣟࣆࢡࣜࣥࡢᢚไ↷ᑕ࡛ㄏⓎࡉࢀࡿ⾲⓶ࡢ⫧ཌᑐࡍࡿࢩࢼࣟࣆࢡࣜࣥࡢᢚไຠᯝຠᯝຠᯝຠᯝ 㸦A㸧 UV ↷ᑕࡋࡓ࣐࢘ࢫ⓶ࡢ HE ᰁⰍ 㸦A,B㸧UV ↷ᑕࡼࡿ⾲⓶㐣ቑṪࡢ CP ࡢຠᯝࠋDBA/2 ࣐࢘ࢫࡣ↓సⅭ 4 ⩌ศࡅࠊྛ⩌ᙜࡓࡾ ᑡ࡞ࡃࡶ7 ༉⏝࠸ࡓࠋCP ᢞ⩌ࡣࠊ462.2 mg/kg㸦ࢩࢼࣟࣆࢡࣜࣥ 3 mg/kg ┦ᙜ㸧ࡲࡓࡣ 92.4 mg/kg 㸦ࢩࢼࣟࣆࢡࣜࣥ0.6 mg/kg ┦ᙜ㸧ࢆ㐃᪥⤒ཱྀᢞࡋࡓࠋUV ↷ᑕ⩌ࢥࣥࢺ࣮ࣟࣝ⩌ࡣࠊ462.2 mg/kg ࡢࢹ࢟ࢫࢺࣜࣥࢆྠᵝᢞࡋࡓࠋࢹ࣮ࢱࡣᖹᆒsS.D.ࠊ᭷ពᕪࡣ ANOVA ࡼࡾ ** p <0.01ࠊ * p <0.05 ♧ࡋࡓࠋscale bar = 20μm.
㸦C㸧࣐࢘ࢫ⓶ࡢ PCNA Ⓨ⌧ࡢචᰁⰍࠋscale bar = 20μm.
ᅗ ᅗ ᅗ ᅗ UVB↷ᑕ↷ᑕ↷ᑕ↷ᑕࡼࡿ࣓ࣛࣀࢧࢺቑṪᑐࡍࡿࢩࢼࣟࣆࢡࣜࣥࡼࡿᢚไຠᯝࡼࡿ࣓ࣛࣀࢧࢺቑṪᑐࡍࡿࢩࢼࣟࣆࢡࣜࣥࡼࡿᢚไຠᯝࡼࡿ࣓ࣛࣀࢧࢺቑṪᑐࡍࡿࢩࢼࣟࣆࢡࣜࣥࡼࡿᢚไຠᯝࡼࡿ࣓ࣛࣀࢧࢺቑṪᑐࡍࡿࢩࢼࣟࣆࢡࣜࣥࡼࡿᢚไຠᯝ 㸦A㸧࣓ࣛࣀࢧࢺࡢL-DOPAᰁⰍ 㸦A,B㸧UV↷ᑕࡼࡿ࣓ࣛࣀࢧࢺቑṪࡢCPࡢຠᯝࠋDBA/2 ࣐࢘ࢫࡣ↓సⅭ 4 ⩌ศࡅࠊྛ⩌ᙜࡓ ࡾᑡ࡞ࡃࡶ7 ༉⏝࠸ࡓࠋCPᢞ⩌ࡣࠊ462.2 mg/kg㸦ࢩࢼࣟࣆࢡࣜࣥ 3 mg/kg┦ᙜ㸧ࡲࡓࡣ 92.4 mg/kg 㸦ࢩࢼࣟࣆࢡࣜࣥ0.6 mg/kg┦ᙜ㸧ࢆ㐃᪥⤒ཱྀᢞࡋࡓࠋUV↷ᑕ⩌ࢥࣥࢺ࣮ࣟࣝ⩌ࡣࠊ462.2 mg/kg ࡢࢹ࢟ࢫࢺࣜࣥࢆྠᵝᢞࡋࡓࠋࢹ࣮ࢱࡣᖹᆒsS.D.ࠊ᭷ពᕪࡣANOVAࡼࡾ ** p<0.01ࠊn.s. ᭷ពᕪ ࡞ࡋ♧ࡋࡓࠋscale bar = 500μm. A B
⪃ ⪃ ⪃ ⪃ ᐹᐹᐹᐹ ⣸እ⥺ࡣ⓶᭷ᐖ࡞⎔ቃせᅉࡢ㸯ࡘ࡛࠶ࡾࠊ⓶⪁ࡢཎᅉ࡛࠶ࡿࡇࡀ▱ࡽࢀ࡚ ࠸ࡿ7,41,42ࠋ⣸እ⥺ࡼࡾ⓶࡛ࡣIL-1ࠊTNF-αྵࡴ⅖ᛶࢧࢺ࢝ࣥ43-46ࡸMMP-1, bFGF ࡀ⏘⏕ࡉࢀࡿ15,16ࠋNF-κB ࡣච⅖ᛂࡢỴᐃⓗᅉᏊ࡛ࠊ⅖ᛶࢧࢺ࡛࢝ࣥ࠶ ࡿIL-1 TNF-αࡀ NF-κB ࢆㄏⓎࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ37ࠋNF-κB ࡣࣞࣝࢠ࣮ᛶ⓶ ⅖ࡸⓄࠊ⓶ࡀࢇࢆྵࡴࡉࡲࡊࡲ࡞⓶ࡶ㛵ಀࡋ12ࠊࡲࡓࠊ⣸እ⥺ࡼࡗ࡚άᛶ ࡉࢀࠊIL-1ࠊTNF-αࠊMMP-1ࠊbFGF ࡞ࡢ㑇ఏᏊⓎ⌧ࢆㄏⓎࡍࡿࡇ࡛⓶ࡢ⪁ࢆ㉳ࡇ ࡍሗ࿌ࡉࢀ࡚࠸ࡿ10,11ࠋNF-κB ࡢ㌿άᛶ㜼ᐖࡣࠊ⣸እ⥺ࡼࡗ࡚ㄏⓎࡉࢀࡿⓗ࡞ኚ ࡸග⪁ࡢቑᝏࢆண㜵ࡍࡿୖ࡛㔜せ࡞ᙺࢆᯝࡓࡋ࡚࠸ࡿ⪃࠼ࡽࢀࡿࠋࡇࢀࡲ࡛᪤ ▱ࡢNF-κB 㜼ᐖ࡛࠶ࡿ parthenolide ࡢ⣸እ⥺ࡼࡾㄏᑟࡉࢀࡿග⪁㜵Ṇస⏝ࡸ NF-κB 㜼ᐖࡢᒁᡤ㐺⏝ࡼࡿග⪁ࡢ㜵Ṇࡀሗ࿌ࡉࢀ࡚࠸ࡿ18,34ࠋ ᮏ◊✲࠾࠸࡚ࠊ࣮ࢸࢳ࣮ࣙࢡࡀຠᯝⓗNF-κB ࡢ㌿άᛶࢆ㜼ᐖࡋࠊග⪁ࣉࣟ ࢭࢫࢆ㜵Ṇ࡛ࡁࡿሗ࿌ࡋࡓࠋࡲࡓࠊ࣮ࢸࢳ࣮ࣙࢡࡢᡂศࡢ㸯ࡘ࡛࠶ࡿࢩࢼࣟࣆࢡࣜ ࣥࡀᢠග⪁స⏝ࢆᣢࡘࡇࡶ☜ㄆࡋࡓࠋࢩࢼࣟࣆࢡࣜࣥࡀNF-κB㸦p65㸧ࡢ㌿άᛶࢆ 㜼ᐖࡋࡓࡇࡽࠊࢩࢼࣟࣆࢡࣜࣥࡢNF-κB 㜼ᐖࡢせ࡞ࢱ࣮ࢤࢵࢺࡣࠊI-κB ࡢಟ㣭ࠊศ ゎࡼࡾࡶୗὶ࠶ࡿ⪃࠼ࡽࢀࡿࠋࡉࡽࠊࢩࢼࣟࣆࢡࣜࣥࡣNF-κB ࡢ᰾⛣⾜ࢆ㜼ᐖࡋࡓࠋ ࡼࡗ࡚ࠊస⏝ᶵᗎࡢ1 ࡘࡋ࡚ࢩࢼࣟࣆࢡࣜࣥࡀάᛶࡉࢀࡓ NF-κB ࡢ᰾⛣⾜ࢆ㜼ᐖࡍࡿ ࡇࡀ♧၀ࡉࢀࡓࠋࡲࡓࠊࢩࢼࣟࣆࢡࣜࣥࡢ⤒ཱྀᢞࡀ⾲⓶ࡢ⫧ཌࡸ࣓ࣛࣀࢧࢺࡢ㐣ቑ Ṫ࠸ࡗࡓ⣸እ⥺ࡼࡾᘬࡁ㉳ࡇࡉࢀࡿ⓶ࡢග⪁ࢆᢚไ࡛ࡁࡿࡇࢆ᫂ࡽࡋࡓࠋ
⤖ ⤖ ⤖ ⤖ㄽㄽㄽㄽ ᮏ◊✲࠾࠸࡚ࠊࢩࢼࣟࣆࢡࣜࣥࡀ⾲⓶ࡢ⫧ཌࡸ࣓ࣛࣀࢧࢺࡢ㐣ቑṪ࠸ࡗࡓග⪁ ࢆᢚไ࡛ࡁࡿࡇࢆ᫂ࡽࡋࡓࠋ⓶ග⪁ࡢࣉࣟࢭࢫࡢᅉࡣNF-κB ࡛࠶ࡿ࠸࠺ࡇ ᇶ࡙ࡁࠊࢩࢼࣟࣆࢡࣜࣥࡢ⤒ཱྀᢞࡀNF-κB ࡢ㌿άᛶࢆ㜼ᐖࡍࡿࡇࡼࡾග⪁ ᑐࡋ࡚ຠᯝⓗ࡞ࡇࢆ⣽⬊ࡸ࣐࢘ࢫࢆ⏝࠸ࡓᐇ㦂ࢆ㏻ࡌ࡚ド࡛᫂ࡁࡓࠋ ࡇࡢ࣮ࢸࢳ࣮ࣙࢡᢳฟ≀ࡣ⢝ရࡸ⨾ᐜ┠ⓗࡢࢧࣉ࣓ࣜࣥࢺ࡞࠸ࡃࡘࡢၟရ ᛂ⏝ࡉࢀ࡚࠾ࡾࠊࢩࢼࣟࣆࢡࣜࣥࡢ⏝ࡼࡿග⪁㜵Ṇࡢ࣓࢝ࢽࢬ࣒ࡀࡉࡽゎ᫂ࡉࢀ ࡿࡇࡀᮇᚅࡉࢀࡿࠋ ㅰ㎡ ㅰ㎡ ㅰ㎡ ㅰ㎡ ✏ࢆ⤊ࢃࡿ⮫ࡳࠊᮏ◊✲ࡢ㐙⾜㝿ࡋ࡚ࠊ⤊ጞᚚᣦᑟࡈ㠴᧡ࢆ㈷ࡾࡲࡋࡓᒱ㜧Ꮫ Ꮫ㝔㐃ྜ⸆་⒪ሗ◊✲⛉⸆⛉Ꮫᑓᨷ ㉥ᑿ ᖾ༤ ᩍᤵᚰࡽឤㅰ࠸ࡓࡋࡲࡍࠋ ᮏ◊✲㝿ࡋࠊpGL3-4kBwt-lucࠊpCMV-p65 ࢆᥦ౪࠸ࡓࡔ࠸ࡓྡྂᒇᕷ❧ᏛᏛ㝔་ Ꮫ◊✲⛉⣽⬊ศᏊ⏕≀Ꮫศ㔝 ᒸᮏ ᑦ ᩍᤵཌࡃᚚ♩⏦ࡋୖࡆࡲࡍࠋ ᮏ◊✲㝿ࡋ⤊ጞ῝࠸ࡈ⌮ゎࢆ࠸ࡓࡔࡁࡲࡋࡓ୍ࣇࣝࢥࢫᰴᘧ♫ Ᏻ⸨ⰾᙪ♫㛗ࠊ ᆤㄔ㛤Ⓨ㒊㛗῝ㅰ⮴ࡋࡲࡍࠋ
ཧ⪃ᩥ⊩ ཧ⪃ᩥ⊩ ཧ⪃ᩥ⊩ ཧ⪃ᩥ⊩
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