実験的歯周炎マウスにおける海馬領域の炎症反応

概要

ㄽᩥࡢ඲ᩥせ⣙



ᐇ㦂ⓗṑ࿘⅖࣐࢘ࢫ࡟࠾ࡅࡿᾏ㤿㡿ᇦࡢ⅖⑕཯ᛂ

ྂ⋢ ኱♸
ᗈᓥ኱Ꮫ኱Ꮫ㝔་ṑ⸆ಖ೺Ꮫ◊✲⛉
༤ኈㄢ⛬ ་ṑ⸆Ꮫᑓᨷ
2019 ᖺᗘ
୺ᣦᑟᩍဨ㸸ᰩཎ ⱥぢ ᩍᤵ
㸦་⣔⛉Ꮫ◊✲⛉ ṑ࿘⑓ែᏛ㸧
1

࠙┠ⓗࠚ
ㄆ▱⑕ࡣグ᠈㞀ᐖࠊぢᙜ㆑㞀ᐖࠊㄆ▱ᶵ⬟㞀ᐖࡸ⾜ືᚰ⌮㞀ᐖࢆక࠺⑕ೃ⩌࡛
࠶ࡿࠋ2030 ᖺ࡟ࡣࠊୡ⏺୰࡛⣙ 7470 ୓ேࡢㄆ▱⑕ᝈ⪅ࡀᏑᅾࡍࡿ࡜ண ࡉࢀ࡚
࠸ࡿࠋ᪥ᮏ࡛ࡣ㧗㱋໬࡟క࠸ࠊㄆ▱⑕ᝈ⪅ᩘࡣቑຍࡋ࡚࠾ࡾࠊ2025 ᖺ࡟ࡣᝈ⪅
ᩘࡣ 700 ୓ே࡟฿㐩ࡋࠊ㧗㱋⪅ࡢ 5 ே࡟ 1 ேࡀㄆ▱⑕ࢆⓎ⑕ࡍࡿ࡜ண ࡉࢀ࡚
࠸ࡿࠋㄆ▱⑕ࡢ 60㹼70㸣ࢆ࢔ࣝࢶࣁ࢖࣐࣮ᆺㄆ▱⑕㸦AD㸧ࡀ༨ࡵࡿ࡜ሗ࿌ࡉࢀ
࡚࠸ࡿࠋAD ࡣᝈ⪅ᮏேࡔࡅ࡛࡞ࡃࠊᐙ᪘ࡸ♫఍ࡢ኱ࡁ࡞ၥ㢟࡛࠶ࡾࠊࡑࡢ࣓࢝
ࢽࢬ࣒ࡢゎ᫂ࠊ἞⒪ἲࡢ㛤Ⓨࡣႚ⥭ࡢㄢ㢟࡛࠶ࡿࠋ
AD ࡣᾏ㤿ࡢⴎ⦰ࠊ⪁ேᩬ㸦࢔࣑ࣟ࢖ࢻ β ࡢỿ╔㸧ࠊ⚄⤒ཎ⥺⥔ኚ໬ࢆ≉ᚩ࡜
ࡋࡓ៏ᛶⓗ࡞⚄⤒ኚᛶ⑌ᝈ࡛࠶ࡾࠊ⦆ࡸ࠿࡟㐍⾜ࡍࡿࠋAD ᝈ⪅ࡢ⬻⤌⧊࡛࢔࣑
ࣟ࢖ࢻ β ࡢỿ╔ࠊ⚄⤒ཎ⥺⥔ኚ໬࡞࡝ࡢ⑓⌮Ꮫⓗኚ໬ࡣ⮫ᗋ⑕≧ࡀ⌧ࢀࡿ 20㹼
30 ᖺ๓࠿ࡽฟ⌧ࡋ࡚࠸ࡿ࡜ሗ࿌ࡉࢀ࡚࠸ࡿࠋṑ࿘⅖ࡢ⨯ᝈᩘࡣ 30㹼40 ṓ௦࡟࠿
ࡅ࡚ⴭࡋࡃቑຍࡍࡿࠋࡇࡢࡇ࡜ࡣࠊ⬻⤌⧊ࡢ⑓⌮Ꮫⓗኚ໬࡜ṑ࿘⅖࡜ࡢ㛫࡟ఱࡽ
࠿ࡢ㛵ಀࡀ࠶ࡿྍ⬟ᛶࢆ♧၀ࡍࡿࠋࡇࡢ㛵㐃ࡀ᫂ࡽ࠿࡟࡞ࢀࡤࠊṑ࿘⑓἞⒪࡟ࡼ
ࡗ࡚ㄆ▱⑕ࡢⓎ⑕ࢆ㐜ࡽࡏࡿࡇ࡜ࡀྍ⬟࡜࡞ࡾࠊࡉࡽ࡟ࠊAD ἞⒪ࡢ᪂つࢱ࣮ࢤ
ࢵࢺࡢ᥈⣴࡟ࡘ࡞ࡀࡿྍ⬟ᛶࡶ࠶ࡿࠋ
 AD ࡢ⑓ែ࡟㛵ࢃࡿᅉᏊࡢ୍ࡘ࡟⬻⤌⧊ࡢ⅖⑕ࡀ࠶ࡿࠋ⬻⤌⧊ෆࡢ⅖⑕ࡣ࢔࣑
ࣟ࢖ࢻ β ࡢ⏘⏕ࠊ⚄⤒ཎ⥺⥔ኚ໬ࠊ⚄⤒⣽⬊㞀ᐖࡸ⣽⬊ṚࢆㄏᑟࡋࠊAD Ⓨ⑕ࡢ
⣲ᅉ࡟࡞ࡿ࡜⪃࠼ࡽࢀ࡚࠸ࡿࠋࡲࡓ⢾ᒀ⑓࡞࡝ࡢ඲㌟ⓗ࡞៏ᛶ⅖⑕࡟ࡼࡿࠊ⾑ὶ
୰ࡢ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥࡢ⏘⏕ࡣ⬻⾑ᾮ㛵㛛㸦BBB㸧ࡢ㏱㐣ᛶࢆஹ㐍ࡋࠊ⬻⤌⧊
ࡢ⅖⑕ࢆᘬࡁ㉳ࡇࡋ࡚࠸ࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋ
ᮏ◊✲࡛ࡣṑ࿘⤌⧊ࡢ⅖⑕ࡀ⬻ࡢ⅖⑕࡟ཬࡰࡍᙳ㡪ࢆゎ᫂ࡍࡿࡇ࡜ࢆ┠ⓗ࡟ࠊ
⤱⣒⤖⣻ṑ࿘⅖ࣔࢹ࣐ࣝ࢘ࢫࡢ⬻ࡢኚ໬ࢆヲ⣽࡟᳨ウࡋࡓࠋ
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࠙ᮦᩱ࡜᪉ἲࠚ
1. ⤱⣒⤖⣻ṑ࿘⅖ࣔࢹࣝ
ᐇ㦂ࡣ 6-8 㐌㱋ࡢ c57BL/6 ࡢ㞤࣐࢘ࢫࢆ⏝࠸ࡓࠋ5-0 ⤱⣒ࢆ⏝࠸࡚ୖ㢡୧ഃ
➨஧⮻ṑ࡟⤖⣻ࡋṑ࿘⅖ࢆⓎ⑕ࡉࡏࡓ⩌ࢆṑ࿘⅖⑕⩌࡜ࡋࡓࠋ⤱⣒ࢆ⤖⣻ࡋ
࡞࠸⩌ࢆṑ࿘೺ᖖ⩌࡜ࡋࡓࠋ
2. ṑ࿘⅖⑕⩌ࡢṑ࿘⤌⧊ࠊ⾑Ύ୰࡟࠾ࡅࡿ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥࡢ᳨ウ
ṑ࿘⅖⑕⩌㸦⤖⣻ 1, 4, 8 㐌㛫㸧࡜ṑ࿘೺ᖖ⩌ࡢṑ⫗୰ࡢ Il6ࠊIl1bࠊTnfa ࡢ
mRNA ࡢⓎ⌧㔞ࢆ real time PCR ࡛ㄪ࡭ࡓࠋ⾑Ύ୰ࡢ IL-1β, TNF-α, IL-6 ࡢ⃰
ᗘࡣ ELISA ࢆ⏝࠸࡚ゎᯒࡋࡓࠋ
3. ᾏ㤿࡟࠾ࡅࡿ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥࡢⓎ⌧
3-1. ⤱⣒⤖⣻㸯㐌㛫ᚋࡢṑ࿘⅖⑕⩌ࡢᾏ㤿ࢆ๗ฟࡋࠊᾏ㤿ࡢ Il6ࠊIl1bࠊTnfa
ࡢ mRNA Ⓨ⌧㔞ࢆ real time PCR ࡛ㄪ࡭ࡓࠋ
3-2. ⾑୰ࡢ IL-6 ࡀᾏ㤿࡟࠾ࡅࡿ Il1b Ⓨ⌧࡟୚࠼ࡿᙳ㡪
ձ

ṑ࿘೺ᖖ⩌࡟ࠊ1000 ng/mice ࡢ recombinant IL-6 ࡲࡓࡣ⏕⌮㣗ሷỈࢆᑿ
㟼⬦࠿ࡽᢞ୚ࡋ㸪ᾏ㤿ࡢ Il1b ࡢ mRNA Ⓨ⌧㔞ࡢኚ໬ࢆㄪ࡭ࡓࠋ

ղ ṑ࿘⅖⑕⩌࡟ࠊ⤱⣒⤖⣻ᙜ᪥ࠊ3 ᪥┠࡟ IL-6 ࡢ୰࿴ᢠయࠊࡲࡓࡣᑐ↷࡜
ࡋ࡚ࣛࢵࢺ IgG ࢆᑿ㟼⬦࠿ࡽᢞ୚ࡋࠊᾏ㤿ࡢ Il1b ࡢ mRNA Ⓨ⌧㔞ࡢኚ໬
ࢆㄪ࡭ࡓࠋ
4. ṑ࿘⅖⑕⩌ࡢᾏ㤿࡟࠾ࡅࡿ BBB ᵓᡂࢱࣥࣃࢡࡢⓎ⌧
4-1. ⤱⣒⤖⣻ 1 㐌㛫ᚋࡢᾏ㤿ࢆ๗ฟࡋࠊᾏ㤿࡟࠾ࡅࡿ tight junction 㛵㐃㑇ఏ
Ꮚ㸦Cldn5, Ocln, Tjp1㸧ࡢ mRNA Ⓨ⌧㔞ࢆ real time PCR ࡛ㄪ࡭ࡓࠋ
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4-2. ⤱⣒⤖⣻ 1 㐌㛫ᚋࡢ඲⬻ࢆ๗ฟࡋࠊ⤌⧊ษ∦ࢆసᡂࡋࠊCD31㸦⾑⟶ෆ
⓶⣽⬊ࡢᣦᶆศᏊ㸧࡜ Cldn5 ࢆ⺯ගⰍ⣲ᶆ㆑ᢠయ࡟ࡼࡿ஧㔜ᰁⰍࢆ⾜࠸ࠊ
⾑⟶ෆ⓶⣽⬊ࡢ Cldn5 Ⓨ⌧ࢆ⺯ග㢧ᚤ㙾ୗ࡛ほᐹࡋࡓࠋ
5. ⾑୰ IL-6 ࡀᾏ㤿⾑⟶ෆ⓶⣽⬊ࡢ Cldn5 Ⓨ⌧࡟ᑐࡍࡿᙳ㡪
ṑ࿘⅖⑕⩌࣐࢘ࢫ࡟ࠊ⤱⣒⤖⣻ᙜ᪥ࠊ3 ᪥ᚋ࡟ IL-6 ࡢ୰࿴ᢠయࡲࡓࡣᑐ↷
࡜ࡋ࡚ࣛࢵࢺ IgG ࢆᑿ㟼⬦࠿ࡽᢞ୚ࡋࠊᢞ୚ 7 ᪥ᚋ࡟඲⬻ࢆ๗ฟࡋࠊᐇ㦂 42 ࡜ྠᵝࡢ᪉ἲ࡛⾑⟶ෆ⓶⣽⬊ࡢ Cldn5 Ⓨ⌧ࢆほᐹࡋࡓࠋ
6. ṑ࿘⅖⑕⩌ࡢ BBB ࡢ㏱㐣ᛶஹ㐍࡟ᑐࡍࡿ᳨ウ
  ṑ࿘೺ᖖ⩌ࠊࡲࡓࡣ⤱⣒⤖⣻㸵᪥┠ࡢṑ࿘⅖⑕⩌࣐࢘ࢫ࡟ biotin ᶆ㆑ 3000D
ࡢ dextran ࢆᑿ㟼⬦࠿ࡽᢞ୚ࡋࠊᢞ୚ 10 ศᚋ࡟඲⬻ࢆ๗ฟࡋࡓࠋ⤌⧊ษ∦ࢆస
ᡂࡋࠊCD31 ࢆ⺯ගⰍ⣲ᶆ㆑ᢠయ࡟ࡼࡿᰁⰍࢆ⾜࠸ࠊdextran ࡀ⾑⟶ෆ࠿ࡽ⬻⤌
⧊ෆ࡬₃ฟࡋ࡚࠸ࡿീࢆほᐹࡋࡓࠋ


࠙⤖ᯝࠚ
1. ṑ࿘⅖ࣔࢹ࣐ࣝ࢘ࢫࡢṑ࿘⤌⧊࡜඲㌟ࡢ⅖⑕࡟ཬࡰࡍᙳ㡪
1) ṑ࿘⅖⑕⩌࡛ࡣṑ⫗⤌⧊୰࡛ Il6ࠊIl1bࠊTnfa ࡢ mRNA Ⓨ⌧ࡀ⤱⣒⤖⣻ᚋ 1
㐌㛫┠࡛᭷ព࡟㧗࠿ࡗࡓࠋ
2) ṑ࿘⅖⑕⩌࡛⾑Ύ୰ࡢ IL-6 ࡢⓎ⌧ࡀ᭷ព࡟㧗࠿ࡗࡓࠋ

2. ṑ࿘⅖⑕⩌ࡀᾏ㤿㡿ᇦࡢ⅖⑕࡟ཬࡰࡍᙳ㡪
1) ṑ࿘⅖⑕⩌ࡣᾏ㤿㡿ᇦ࡛ Il1b ࡢ mRNA Ⓨ⌧ࡀ᭷ព࡟㧗࠿ࡗࡓࠋ
4

2) recombinant Murine IL-6 ࡢ⾑୰ᢞ୚ࡣᾏ㤿㡿ᇦ࡛ Il1b ࡢ mRNA Ⓨ⌧ࢆୖ᪼
ࡉࡏࡓࠋ
3) anti-mouse IL-6 antibody ࡢ⾑୰ᢞ୚ࡣࠊṑ࿘⅖⑕⩌࡛⏕ࡌࡿᾏ㤿㡿ᇦࡢ Il1b
ࡢ mRNA Ⓨ⌧ࢆᢚไࡋࡓࠋ

3. ṑ࿘⅖ࡀᾏ㤿㡿ᇦ࡟࠾ࡅࡿ⾑ᾮ⬻㛵㛛㸦Blood-Brain Barrier : BBB㸧࡟ཬࡰࡍ
ᙳ㡪
1) ṑ࿘⅖⑕⩌࡛ᾏ㤿㡿ᇦࡢ Cldn5 ࡢⓎ⌧㔞ࡣప࠿ࡗࡓࠋ
2) anti-mouse IL-6 antibody ࡢ⾑୰ᢞ୚ࡣࠊṑ࿘⅖⑕⩌࡛⏕ࡌࡿᾏ㤿㡿ᇦࡢ Cldn5
ࡢⓎ⌧㔞పୗࢆᢚไࡋࡓࠋ
3) ṑ࿘⅖⑕⩌࡛᭷ព࡟⾑⟶እ࡬⺯ග࣭biotin ᶆ㆑ 3,000D ࡢ⺯ගࢆⓎࡍࡿ㠃✚
ࡀᗈ࠿ࡗࡓࠋ

࠙⪃ᐹࠚ
ᮏ◊✲ࡢ⤖ᯝ࠿ࡽṑ࿘⤌⧊ࡢ⅖⑕࡟ࡼࡗ࡚ㄏᑟࡉࢀࡓ⾑୰ࡢ IL-6 ࡟ࡼࡗ࡚ࠊ
ᾏ㤿㡿ᇦࡢ⾑⟶ෆ⓶⣽⬊⭷ୖࡢ Cldn5 ࡢⓎ⌧ࢆపୗࡉࡏࠊBBB ࡢ㏱㐣ᛶࡀஹ㐍
ࡋࡓࠋࡲࡓࠊࡇࢀ࡟ࡼࡾᾏ㤿㡿ᇦ࡛⅖⑕཯ᛂࡀㄏᑟࡉࢀࡓࠋ
ṑ࿘⅖ࢆ᪩ᮇ࡟἞⒪ࡋ៏ᛶ⅖⑕ࢆྲྀࡾ㝖ࡃࡇ࡜࡟ࡼࡗ࡚ࠊᾏ㤿㡿ᇦࡢ⅖⑕཯
ᛂࢆᢚไฟ᮶ࢀࡤࠊAD Ⓨ⑕᫬ᮇࢆ㐜ࡽࡏࡿࡇ࡜ࡀฟ᮶ࠊ┦ᑐⓗ࡞ AD ᝈ⪅ᩘࡢ
ῶᑡ࡟⧅ࡀࡿྍ⬟ᛶࡀ࠶ࡿ࡜⪃࠼ࡿࠋ

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