Synergistic effect of collagen and CXCL12 in the low doses on human platelet activation( 要約版(Digest) )

Title Synergistic effect of collagen and CXCL12 in the low doses on human platelet activation( 要約版(Digest) )

Author(s) 中島, 大樹

Report No.(Doctoral

Degree) 博士(医学) 甲第1156号

Issue Date 2021-03-25

Type 博士論文

Version none

URL http://hdl.handle.net/20.500.12099/81565

※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。

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Ꮫ఩ㄽᩥせ⣙

Extended Summary in Lieu of the Full Text of a Doctoral Thesis

Ặ ྡ㸸 Full Name

୰ ᓥ ኱ ᶞ

Ꮫ఩ㄽᩥ㢟┠㸸 Thesis Title

6\QHUJLVWLFHIIHFWRIFROODJHQDQG&;&/LQWKHORZGRVHVRQKXPDQ SODWHOHWDFWLYDWLRQ

Ꮫ఩ㄽᩥせ⣙㸸 Summary of Thesis

CXCL12ࡣCXCࣇ࢓࣑࣮ࣜ࡟ศ㢮ࡉࢀࡿࢣࣔ࢝࢖ࣥࡢ୍ࡘ࡛࠶ࡾ㸪ࡑࡢ≉␗ⓗཷᐜయ࡛࠶ࡿCX

CR4࠾ࡼࡧCXCR7ࢆ௓ࡋస⏝ࡍࡿࠋCXCL12ࡣⓑ⾑⌫ࡢ⅖⑕㒊఩࡬ࡢ㐟㉮ㄏᑟ࡟ຍ࠼㸪㐀⾑㸪⬇Ⓨ

⏕࠾ࡼࡧ⾑⟶᪂⏕࡞࡝ࡢ⏕⌮ⓗࣉࣟࢭࢫ࡟㛵୚ࡋ࡚࠸ࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋ㐀⾑ᶵᵓ㸪≉࡟⾑ᑠ ᯈᙧᡂ࡟࠾࠸࡚㸪CXCL12ࡣ㸪ᕧ᰾ⱆ⌫ࡢᕧ᰾⌫࡬ࡢศ໬࠾ࡼࡧᡂ⇍⾑ᑠᯈࡢ⾑୰࡬ࡢㄏᑟ࡟ᚲ㡲

࡛࠶ࡿࡇ࡜ࡀ᫂ࡽ࠿࡟ࡉࢀ࡚࠸ࡿࠋ

⾑ᑠᯈࡣṆ⾑࡟࠾࠸࡚୰ᚰⓗ࡞ᙺ๭ࢆᯝࡓࡋ࡚࠸ࡿࠋ⾑⟶ෆ⓶ࡢ㞀ᐖ࡟ࡼࡾ㟢ฟࡋࡓࢥ࣮ࣛࢤ

ࣥࡣ㸪Glycoprotein㸦GP㸧VI࠾ࡼࡧȘ2ș1࢖ࣥࢸࢢࣜࣥ➼ࡢࢥ࣮ࣛࢤࣥཷᐜయࢆ௓ࡋ࡚⾑ᑠᯈࡢจ 㞟ࢆಁ㐍ࡍࡿࠋ⾑ᑠᯈࡢจ㞟࡟కࡗ࡚㸪⣽⬊ቑṪᅉᏊ࡛࠶ࡿplatelet-derived growth factor AB㸦PD GF-AB㸧ࡢศἪ࠾ࡼࡧ⅖⑕ច㉳≀㉁࡛࠶ࡿsoluble CD40 ligand㸦sCD40L㸧ࡢ㐟㞳ࡀច㉳ࡉࢀࡿࠋ

୍᪉㸪⾑ᑠᯈᶵ⬟ࡢஹ㐍ࡣෙື⬦ཬࡧ⬻⾑⟶➼ࡢ⾑ᰦᛶ࢖࣋ࣥࢺࡢࢺ࣮ࣜ࢞࡜࡞ࡿ࡜⪃࠼ࡽࢀ࡚

࠸ࡿࠋ⏦ㄳ⪅ࡽࡢ◊✲ᐊ࡛ࡣ㸪ࢥ࣮ࣛࢤࣥࡣp38 mitogen-activated protein kinase㸦MAPK㸧࠾ࡼࡧ p44/p42 MAPKࡢάᛶ໬ࢆ௓ࡋ㸪⾑ᑠᯈᶵ⬟ࢆάᛶ໬ࡍࡿࡇ࡜ࢆ᫂ࡽ࠿࡟ࡋ࡚࠸ࡿࠋ

ࢫࢺࣞࢫࢱࣥࣃࢡ㉁㸦Heat Shock Protein㸸HSP㸧ࡣࢫࢺࣞࢫᛂ⟅ࡢ㝿࡟ㄏᑟࡉࢀࡿࢱࣥࣃࢡ㉁

ࡢ⥲⛠࡛࠶ࡾ㸪ศᏊࢩࣕ࣌ࣟࣥ࡜ࡋ࡚ᶵ⬟ࡍࡿ࡜⪃࠼ࡽࢀ࡚࠸ࡿࠋHSP27ࡣపศᏊ㔞HSPࡢ୍ࡘ࡛

࠶ࡾ㸪⾑ᑠᯈࢆྵࡴᵝࠎ࡞⣽⬊࡟ᜏᖖⓗ࡟Ⓨ⌧ࡋ࡚࠸ࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋࡲࡓ㸪HSP27ࡣࣜࣥ

㓟໬࡟ࡼࡿ⩻ヂᚋಟ㣭ࢆཷࡅࡿࡇ࡜ࡀ▱ࡽࢀ࡚࠸ࡿࠋ㏆ᖺ㸪HSP27ࡣ⣽⬊ෆࡢࡳ࡞ࡽࡎ㸪⣽⬊እ࡛

⅖⑕཯ᛂࡢไᚚᅉᏊࡢ୍ࡘ࡜ࡋ࡚ࡶᶵ⬟ࡍࡿࡇ࡜ࡀ᫂ࡽ࠿࡟ࡉࢀ࡚࠸ࡿࠋ⏦ㄳ⪅ࡽࡢ◊✲ᐊ࡛ࡣ㸪 ࢥ࣮ࣛࢤ่⃭ࣥ࡟ࡼࡿPDGF-ABࡢศἪ࠾ࡼࡧsCD40Lࡢ㐟㞳࡟కࡗ࡚㸪p44/p42 MAPKࢆ௓ࡋࡓHS P27ࡢࣜࣥ㓟໬ࡀច㉳ࡉࢀࡿࡇ࡜㸪ࡲࡓ㸪⢾ᒀ⑓ᝈ⪅࡟࠾࠸࡚㸪HSP27ࡣࢥ࣮ࣛࢤ่⃭ࣥ࡟ࡼࡗ࡚

ࣜࣥ㓟໬ࡉࢀࡓᚋ㸪⾑ᑠᯈ࠿ࡽ⾑₢୰࡬㐟㞳ࡉࢀࡿࡇ࡜ࢆ᫂ࡽ࠿࡟ࡋ࡚࠸ࡿࠋ

ࣄࢺᡂ⇍⾑ᑠᯈ࡟࠾࠸࡚CXCL12ࡣᙅ࠸⾑ᑠᯈจ㞟ࢆ⏕ࡌࡿప⏝㔞ࡢࢥ࣮ࣛࢤ่⃭ࣥ࡟ࡼࡿ⾑ᑠ ᯈจ㞟స⏝ࢆቑᙉࡍࡿࡇ࡜ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋࡋ࠿ࡋ㸪CXCL12ࡢࣄࢺᡂ⇍⾑ᑠᯈ࡟ᑐࡍࡿస⏝ࡢ ヲ⣽ࡣ㸪ᮍࡔ᫂ࡽ࠿࡛ࡣ࡞࠸ࠋᮏ◊✲࡟࠾࠸࡚㸪ప⏝㔞ࢥ࣮ࣛࢤࣥ࡟ࡼࡿࣄࢺ⾑ᑠᯈస⏝࡟ᑐࡍࡿ

CXCL12ࡢᙳ㡪ࢆ᫂ࡽ࠿࡟ࡍࡿࡇ࡜ࢆ┠ⓗ࡜ࡋ㸪ゎᯒࢆ⾜ࡗࡓࠋ

࠙ᑐ㇟࡜᪉ἲࠚ

೺ᖖ⪅࠿ࡽ㟼⬦⾑ࢆ᥇ྲྀࡋ㸪㐲ᚰศ㞳ἲ࡟ࡼࡾከ⾑ᑠᯈ⾑₢㸦PRP㸧ࢆㄪᩚࡋࡓࠋPRPࢆCXCR4

୰࿴ᢠయ㸪CXCR7୰࿴ᢠయ࠾ࡼࡧSB203580㸦p38 MAPK 㜼ᐖ⸆㸧࡛๓ฎ⌮ࡋࡓࡢࡕ࡟㸪ࢥ࣮ࣛࢤ

ࣥ㸪CXCL12㸪ࢺࣟࣥࣅࣥཷᐜయάᛶ໬࣌ࣉࢳࢻ㸦thrombin receptor-activating peptide: TRAP㸧㸪A DP࠾ࡼࡧࢥࣥࣂࣝ࢟ࢩࣥ㸦㑅ᢥⓗGPVI࢔ࢦࢽࢫࢺ㸧่࡛⃭ࡋ㸪ග㏱㐣ἲཬࡧ࣮ࣞࢨ࣮ගᩓ஘ࢩࢫ ࢸ࣒ࢆ⏝࠸࡚⾑ᑠᯈจ㞟ࢆ ᐃࡋࡓࠋ⾑ᑠᯈ࠿ࡽࡢPDGF-ABࡢศἪ㸪sCD40Lࡢ㐟㞳࠾ࡼࡧࣜࣥ㓟

໬HSP27ࡢ㐟㞳ࡣELISAἲ࡛ ᐃࡋࡓࠋ࢚࢘ࢫࢱࣥࣈࣟࢵࢺἲ࡟ࡼࡾp38 MAPK㸪p44/p42 MAPK

࠾ࡼࡧHSP27ࡢࣜࣥ㓟໬ࣞ࣋ࣝࢆゎᯒࡋࡓࠋ

࠙⤖ᯝࠚ

1㸧ప⏝㔞ࢥ࣮ࣛࢤࣥ㸦0.2 ȣg/ml㸧࡜ప⏝㔞CXCL12㸦10 ng/ml㸧ࡣ㸪ࡑࢀࡒࢀ༢⊂࡛ࡣ㸪࡯࡜ࢇ

࡝⾑ᑠᯈจ㞟࡟ᙳ㡪ࢆཬࡰࡉ࡞࠿ࡗࡓࡀ㸪ྠ᫬่⃭࡟ࡼࡗ࡚᭷ព࡟⾑ᑠᯈจ㞟ࢆಁ㐍ࡋࡓࠋ୍᪉㸪 ప⏝㔞TRAP㸦7 ȣM㸧࠶ࡿ࠸ࡣప⏝㔞ADP㸦0.5 ȣM㸧࡜ప⏝㔞CXCL12㸦10 ng/ml㸧ࡢྠ᫬่⃭

࡛ࡣ⾑ᑠᯈจ㞟࡟ఱࡽᙳ㡪ࢆཬࡰࡉ࡞࠿ࡗࡓࠋ

2㸧ప⏝㔞ࢥࣥࣂࣝ࢟ࢩࣥ㸦20 ng/ml㸧࡜ప⏝㔞CXCL12㸦10 ng/ml㸧ࡣ㸪ࡑࢀࡒࢀ༢⊂࡛ࡣ㸪࡯࡜

ࢇ࡝⾑ᑠᯈจ㞟࡟ᙳ㡪ࢆཬࡰࡉ࡞࠿ࡗࡓࡀ㸪ྠ᫬่⃭࡟ࡼࡗ࡚⾑ᑠᯈจ㞟ࢆಁ㐍ࡋࡓࠋ

3㸧ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CXCL12ࡢྠ᫬่⃭࡟ࡼࡗ࡚ಁ㐍ࡉࢀࡓ⾑ᑠᯈจ㞟ࡣCXCR4୰࿴ᢠ య࡟ࡼࡗ࡚᭷ព࡟ᢚไࡉࢀࡓࡀ㸪CXCR7୰࿴ᢠయࡣఱࡽᙳ㡪ࢆཬࡰࡉ࡞࠿ࡗࡓࠋ

4㸧ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CXCL12ࡣ㸪ࡑࢀࡒࢀ༢⊂࡛ࡣPDGF-ABศἪ࠾ࡼࡧsCD40L㐟㞳࡟ఱ

ࡽᙳ㡪ࢆཬࡰࡉ࡞࠿ࡗࡓࡀ㸪ྠ᫬่⃭࡟ࡼࡗ࡚᭷ព࡟PDGF-ABศἪ࠾ࡼࡧsCD40L㐟㞳ࢆಁ㐍ࡋࡓࠋ 5㸧ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CXCL12ࡣ㸪ࡑࢀࡒࢀ༢⊂࡛ࡣp44/p42 MAPK࠾ࡼࡧp38 MAPKࡢࣜ

ࣥ㓟໬࡟ఱࡽᙳ㡪ࢆཬࡰࡉ࡞࠿ࡗࡓࠋ୍᪉㸪ྠ᫬่⃭࡟ࡼࡗ࡚p44/p42 MAPKࡢࣜࣥ㓟໬࡟ࡣఱࡽ

ᙳ㡪ࢆཬࡰࡉ࡞࠿ࡗࡓࡀ㸪p38 MAPKࡢࣜࣥ㓟໬ࢆಁ㐍ࡋࡓࠋ

6㸧SB203580࡟ࡼࡾ㸪ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CXCL12ࡢྠ᫬่⃭࡟ࡼࡗ࡚ಁ㐍ࡉࢀࡓ⾑ᑠᯈจ 㞟㸪p38 MAPKࡢࣜࣥ㓟໬㸪PDGF-ABศἪ࠾ࡼࡧsCD40L㐟㞳ࡣ᭷ព࡟ᢚไࡉࢀࡓࠋ

7㸧ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CXCL12ࡣ㸪ࡑࢀࡒࢀ༢⊂࡛ࡣHSP27ࡢࣜࣥ㓟໬࡟ఱࡽᙳ㡪ࢆཬࡰ

ࡉ࡞࠿ࡗࡓࡀ㸪ྠ᫬่⃭࡟ࡼࡗ࡚HSP27ࡢࣜࣥ㓟໬ࢆಁ㐍ࡋࡓࠋ

8㸧SB203580࡟ࡼࡾ㸪ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CXCL12ࡢྠ᫬่⃭࡟ࡼࡗ࡚ಁ㐍ࡉࢀࡓHSP27ࡢ

ࣜࣥ㓟໬࠾ࡼࡧࣜࣥ㓟໬HSP27㐟㞳ࡣ᭷ព࡟ᢚไࡉࢀࡓࠋ

࠙⪃ᐹࠚ

ప⏝㔞ࢥ࣮ࣛࢤࣥ࡞࠸ࡋప⏝㔞ࢥࣥࣂࣝ࢟ࢩࣥ࡜ప⏝㔞CXCL12ࡢྠ᫬่⃭࡟ࡼࡗ࡚⾑ᑠᯈจ 㞟ࡀಁ㐍ࡉࢀ㸪CXCR4୰࿴ᢠయ࡟ࡼࡗ࡚㸪ࡑࡢ⾑ᑠᯈจ㞟ࡀᢚไࡉࢀࡓࡇ࡜࠿ࡽ㸪ప⏝㔞ࢥ࣮ࣛ

ࢤࣥ࡜ప⏝㔞CXCL12ࡢྠ᫬่⃭࡟ࡼࡿ⾑ᑠᯈจ㞟ࡣ㸪ཷᐜయ࡜ࡋ࡚ࡑࢀࡒࢀGPVI࡜CXCR4ࢆ௓

ࡋ࡚࠸ࡿࡇ࡜ࡀ♧၀ࡉࢀࡓࠋࡲࡓ㸪ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CXCL12ࡢྠ᫬่⃭ࡣp38 MAPKࡢ

ࣜࣥ㓟໬ࢆಁ㐍ࡍࡿࡇ࡜㸪ࡲࡓ㸪SB203580࡟ࡼࡗ࡚㸪⾑ᑠᯈจ㞟㸪PDGF-ABศἪ㸪sCD40L㐟㞳㸪 HSP27ࣜࣥ㓟໬࠾ࡼࡧࣜࣥ㓟໬HSP27㐟㞳ࡀᢚไࡉࢀࡓࡇ࡜࠿ࡽ㸪ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CX CL12ࡣ㸪p38 MAPKࡢάᛶ໬ࢆ௓ࡋ࡚⾑ᑠᯈᶵ⬟ࢆಁ㐍ࡋ࡚࠸ࡿࡇ࡜ࡀ♧၀ࡉࢀࡓࠋᮏ◊✲࡛♧

ࡉࢀࡓ⾑ᑠᯈάᛶ໬ᶵᗎࡣ㸪⾑୰CXCL12ࣞ࣋ࣝࡀ㧗್ࡢᝈ⪅࡟࠾ࡅࡿ⾑ᰦᙧᡂࡢᶵᗎࡢ୍ࡘ࡜⪃

࠼ࡽࢀ㸪⾑ᰦ⑕࡟ᑐࡍࡿ᪂ࡓ࡞἞⒪㛤Ⓨࡢ୍ຓ࡜࡞ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋ

࠙⤖ㄽࠚ

ࣄࢺ⾑ᑠᯈ࡟࠾࠸࡚㸪ప⏝㔞ࢥ࣮ࣛࢤࣥ࡜ప⏝㔞CXCL12ࡣ㸪⾑ᑠᯈจ㞟㸪PDGF-ABศἪ㸪sCD40L 㐟㞳㸪HSP27ࣜࣥ㓟໬࠾ࡼࡧࣜࣥ㓟໬HSP27㐟㞳ࢆ㸪p38 MAPKࡢάᛶ໬ࢆ௓ࡋ࡚┦஌ⓗ࡟ಁ㐍ࡍ

ࡿࡇ࡜ࡀ♧၀ࡉࢀࡓࠋ