ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠᾏ⸴⏤᮶⬡㉁࡟࠾ࡅࡿᢠ⅖⑕స⏝ࡢゎ᫂࡜ࡑࡢά⏝ ➨  ሗ ࢫࢧࣅࣀࣜ࡟࠾ࡅࡿᢠ⅖⑕ᛶ⬡㉁ࡢ≉ᐃ㣗ရᕤᴗ㒊
ᰄ᳜ᆂ௓ 㭯⏣⿱⨾ బ⸨┿బᜨྜྷᮧ⮧ྐ  ᮏ◊✲࡛ࡣ㸪ᶵ⬟ᛶ⣲ᮦ࡜ࡋ࡚ࡢࢫࢧࣅࣀࣜࡢ⏝㏵㛤Ⓨࢆᚿྥࡋ࡚㸪ᢠ⅖⑕స⏝ࢆ᭷ࡍࡿࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁ࡢ≉ᐃ໬ࢆヨࡳࡓ㸬࣐࢘ࢫ⏤᮶ RAW264.7 ⣽⬊ࢆ኱⭠⳦⏤᮶LPS ่࡛⃭ࡋ࡚ NO ཬࡧࢧ࢖ࢺ࢝࢖ࣥ⏘⏕ࢆㄏᑟࡉࡏ㸪ࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮࡟ࡼࡾศ㞳⢭〇ࡋࡓࣜࣥ⬡㉁ࡢඹᏑ࡟ࡼࡿ⏘⏕ᢚไస⏝㸦ᢠ⅖⑕స⏝㸧ࢆホ౯ࡋࡓ㸬ࡑࡢ⤖ᯝ㸪㉁㔞ศᯒ⿦⨨㸦MS㸧࡟ࡼࡾ m/z 783.4 ᳨࡛ฟࡉࢀࡿࣜࣥ⬡㉁࡟㸪ᙉ࠸ NO ཬࡧࢧ࢖ࢺ࢝࢖ࣥࡢ⏘⏕ᢚไࡀㄆࡵࡽࢀࡓ㸬⬡⫫㓟⤌ᡂศᯒ࡞ࡽࡧ࡟ phospholipase A2 ᾘ໬≀ࡢ㉁㔞ศᯒ࡟ࡼࡾ㸪ࡇࡢࣜࣥ⬡㉁ࡀ㸪sn-1 ఩࡟ eicosapentaenoic acid ࡀ㸪sn-2 ఩࡟ 3-hydroxypalmitic acid ࡀࡑࢀࡒ ࢀ ࢚ ࢫ ࢸ ࣝ ⤖ ྜ ࡋ ࡓ 1-eicosapentaenoyl-2-(3-hydroxypalmitoyl)-phosphatidylglycerol(EpPOH-PG) ࡛࠶ࡿࡇ࡜ࢆ᫂ࡽ࠿࡟ࡋࡓ㸬ᵓ㐀ࡢ␗࡞ࡿ✀ࠎࡢࣜࣥ⬡㉁ࡢᢠ⅖⑕స⏝ࢆẚ
㍑ࡋ㸪ᵓ㐀㸭ᶵ⬟┦㛵ࢆ᳨ドࡋࡓ㸬ࣜࣥ⬡㉁ࢡࣛࢫࡢ୰࡛ࡣ㸪phosphatidylglycerol ࡟ᙉ࠸
ᢠ⅖⑕స⏝ࢆㄆࡵ㸪୙㣬࿴⬡⫫㓟ࡀ⤖ྜࡋࡓࣜࣥ⬡㉁ࡢ᪉ࡀ㸪ࡼࡾస⏝ࡀᙉ࠸ഴྥ࡛࠶ࡗ
ࡓ㸬ࡲࡓ㸪ẚ㍑ࡋࡓࣜࣥ⬡㉁ࡢ࠺ࡕ㸪EpPOH-PG ࡀ᭱ࡶᢠ⅖⑕స⏝ࡀᙉ࠿ࡗࡓ㸬௨ୖࡢ⤖ᯝࡣ㸪ࢫࢧࣅࣀࣜ⏤᮶ EpPOH-PG ࡀ㸪ᢠ⅖⑕ᛶࣜࣥ⬡㉁࡜ࡋ࡚ࡢᵓ㐀ⓗ≉ᚩࢆഛ࠼࡚࠾ࡾ㸪
⅖⑕ࡢ㜵Ṇ࡟ᙺ❧ࡘᶵ⬟ᛶ⣲ᮦ࡜ࡋ࡚ࡢྍ⬟ᛶࢆ᫂♧ࡍࡿࡶࡢ࡛࠶ࡿ㸬㸯㸬ࡣࡌࡵ࡟
⅖⑕ࡣ㸪⑓ཎ⳦ࡢឤᰁ➼࡟ࡼࡾ⏕యࡀෆእ࠿ࡽ᭷
ᐖ࡞่⃭ࢆཷࡅࡓ࡜ࡁ࡟⏕ࡌࡿ㜵ᚚ཯ᛂ࡛࠶ࡿ㸬ࡑ
ࡢⓎື࡟㔜せ࡞ࡢࡣ㸪ࠕ⮬↛ච␿ࠖ࡜࿧ࡤࢀࡿ㸪⏕య࡟ࡼࡿ␗≀ࡢ≉␗ⓗㄆ㆑ᶵᵓ࡛࠶ࡿ㸬࣐ࢡࣟࣇ࢓࣮
ࢪࡣ⮬↛ච␿࡟㛵ࢃࡿከᶵ⬟ᛶࡢⓑ⾑⌫࡛㸪⏕య࡟౵ධࡋ࡚ࡁࡓ␗≀ࢆㄆ㆑࣭᤼㝖ࡍࡿࢭࣥࢧ࣮࡜ࡋ࡚
ࡣࡓࡽࡃ㸬౛࠼ࡤ㸪ࢢ࣒ࣛ㝜ᛶ⳦⣽⬊ቨࡢᵓᡂᡂศ
࡛࠶ࡿ࣏ࣜከ⢾㸦LPS㸧ࡣ㸪࣐ࢡࣟࣇ࢓࣮ࢪ࡟ࡼࡾ
ㄆ㆑ࡉࢀ㸪୍㓟໬❅⣲㸦NO㸧㸪ࣉࣟࢫࢱࢢࣛࣥࢪࣥ
࡞࡝ࡢ⏕⌮άᛶ≀㉁ࡸ㸪✀ࠎࡢ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥ
࡞࡝ࡢ⅖⑕ᛶ࣓ࢹ࢕࢚࣮ࢱ࣮ࡢⓎ⌧ㄏᑟࢆಁࡍࡇ࡜
1-3)ࡀ▱ࡽࢀ࡚࠸ࡿ㸬㏆ᖺ㸪ࡀࢇࡸ⏕ά⩦័⑓࡞࡝ࡢ៏ᛶ⑌ᝈࡢ⑓ែᙧ
ᡂ࡟㸪
ࠕ៏ᛶ⅖⑕ࠖࡀ῝ࡃ㛵ࢃࡗ࡚࠸ࡿࡇ࡜ࡀὀ┠ࡉࢀ࡚࠸ࡿ 4)㸬៏ᛶ⅖⑕࡛ࡣ㸪㓟໬ࢫࢺࣞࢫ࡞࡝ࡢ่
⃭ࢆཷࡅࡓ⮚ჾࡢ⣽⬊࠿ࡽᨺฟࡉࢀࡿ⮬ᕫ⏤᮶ࡢᡂศ㸦ෆᅉᛶࣜ࢞ࣥࢻ㸧ࡀ㸪࣐ࢡࣟࣇ࢓࣮ࢪ➼ࡢ⣽⬊
࡟Ⓨ⌧ࡍࡿ Toll ᵝཷᐜయ㸦TLR㸧࡟ࡼࡾㄆ㆑ࡉࢀ࡚㸪ᜏᖖⓗ࡞⅖⑕཯ᛂࢆㄏᑟࡍࡿ࡜⪃࠼ࡽࢀ࡚࠸ࡿ㸬
TLR ࢆ௓ࡋࡓ࣐ࢡࣟࣇ࢓࣮ࢪࡢάᛶ໬࡟ᙳ㡪ࢆཬ
ࡰࡍᵝࠎ࡞እᅉᛶ/ෆᅉᛶࣜ࢞ࣥࢻࡀぢࡘ࠿ࡗ࡚࠾ࡾ 5)㸪ࡑࢀࡽࡣච␿ᶵ⬟ࡢไᚚ࡟ࡣࡓࡽࡃ㣗ရࡸ་⸆㸪࠶ࡿ࠸ࡣච␿⣔ࡢ࣓࢝ࢽࢬ࣒ࢆゎ᫂ࡍࡿࡓࡵࡢ

ࢶ࣮ࣝ࡜ࡋ࡚ࡢά⏝ࡀᮇᚅࡉࢀ࡚࠸ࡿ㸬ᾏ⸴㢮ࡣ㸪࢔ࢪ࢔ᆅᇦ࡟࠾࠸࡚㛗࠸㣗ࡢṔྐࢆ᭷ࡋ㸪㣗⏝ࡢࡳ࡞ࡽࡎ㸪໬⢝ရࡸ㣫⫧ᩱ㸪ቑ⢓๣࣭ࢤࣝ໬๣࡜ࡋ࡚ࡶᖜᗈࡃ฼⏝ࡉࢀ࡚࠸ࡿ㸬᭱㏆࡛ࡣ㸪⏕ά⩦័⑓ࡢண㜵ຠᯝ࡞࡝㸪ᾏ⸴ᡂศࡢ೺ᗣ⥔ᣢᶵ⬟ ࡀ ὀ ┠ ࡉ ࢀ ࡚ ࠸ ࡿ 㸬 ࢫ ࢧ ࣅ ࣀ ࣜ 㸦 Porphyra
yezoensis㸧ࡣ⣚⸴㢮࡟ᒓࡍࡿᾏ⸴ࡢ୍✀࡛㸪ᖸ‶ࡢᕪࡀ⃭ࡋ࠸ᒾ♋ᆅᖏ࡟⮬⏕ࡍࡿࡀ㸪᭷᫂ᾏࢆጞࡵ࡜
ࡍࡿᰤ㣴ሷࡀ㇏ᐩ࡞ᖸ₲ࡸ‴ෆ࡛┒ࢇ࡟㣴Ṫࡉࢀ࡚࠾ࡾ㸪᪥ᮏᅜෆ࡟࠾ࡅࡿᖺ㛫ࡢ⏕⏘㔞ࡣ 35 ୓ࢺࣥ࡟㐩ࡍࡿ 6)㸬
ࡇࢀࡲ࡛㸪ࢫࢧࣅࣀࣜࢆᵓᡂࡍࡿᡂศࡢᦤྲྀ࡟ࡼ
ࡿᵝࠎ࡞⏕⌮స⏝ࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸬ᡃࠎࡣࡇࢀࡲ
࡛࡟㸪ࢫࢧࣅࣀࣜ⏤᮶」ྜ⬡㉁࡟㸪LPS ่⃭࡟ࡼࡿ࣐ࢡࣟࣇ࢓࣮ࢪࡢάᛶ໬ࢆᢚ࠼ࡿస⏝㸦ᢠ⅖⑕స⏝㸧ࡀ࠶ࡿࡇ࡜ࢆ᫂ࡽ࠿࡟ࡋࡓ 7)㸬ࢫࢧࣅࣀࣜ⏤᮶」ྜ⬡㉁ࡣ㸪࣐࢘ࢫ⏤᮶ RAW264.7 ⣽⬊࡟࠾ࡅࡿ LPS ㄏᑟᆺάᛶ໬ࢆᢚ࠼㸪iNOS, COX-2 ࡞ࡽࡧ࡟ TNF-DࡢⓎ⌧ㄏᑟࢆᢚไࡍࡿ㸬ࡲࡓ㸪ࡇࡢ⏬ศࡣ୺࡜ࡋ࡚ࣜࣥ⬡㉁࡜⢾⬡㉁࡛ᵓᡂࡉࢀ㸪ࡑࡢ࠺ࡕࣜࣥ⬡㉁⏬ศ࡟ࡣ phosphatidylglycerol (PG)㸪phosphatidylethanolamine (PE)㸪phosphatidylcholine (PC) ࡀྵࡲࢀࡿࡇ࡜
ࡀ♧၀ࡉࢀ࡚࠸ࡿ㸬Araki ࡽࡢ◊✲ 8),9)࡟ࡼࢀࡤ㸪ࢫࢧࣅࣀࣜ⬡㉁ࡢ⬡⫫㓟⤌ᡂࡣ㸪palmitic acid ୪ࡧ࡟Ⅳ䋭 㪈㪐 䋭

ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠ

⣲ᩘ 20 ࡢ୙㣬࿴⬡⫫㓟ࡀ࡯࡜ࢇ࡝ࢆ༨ࡵ㸪୰࡛ࡶ㨶Ἔ࡟ከࡃྵࡲࢀࡿ eicosapentaenoic acid㸦EPA㸧ࡀ⥲⬡⫫㓟ࡢ 50%௨ୖࢆ༨ࡵࡿ 10)኱ኚࣘࢽ࣮ࢡ࡞ࡶࡢ࡛
࠶ࡿ㸬ࡋ࠿ࡋ࡞ࡀࡽ㸪ࢫࢧࣅࣀࣜ」ྜ⬡㉁⏤᮶ࡢᢠ
⅖⑕స⏝ᛶ⬡㉁ࡢ≉ᐃ໬ࡸ㸪ࡑࡢᵓ㐀Ỵᐃ࡟ࡘ࠸࡚
ࡣ࠸ࡲࡔ㐍ᒎࡀぢࡽࢀ࡞࠸㸬ࡇࡢࡼ࠺࡞⫼ᬒ࠿ࡽ㸪ᮏ◊✲࡛ࡣ㸪ࢫࢧࣅࣀࣜ⏤᮶」ྜ⬡㉁⏬ศ࡟ྵࡲࢀࡿᢠ⅖⑕ᛶ⬡㉁ࡢ≉ᐃ໬ࢆᚿྥࡋ㸪」ྜ⬡㉁ࡢศ⏬࣭⢭〇࡜ᢠ⅖⑕స⏝ࡢホ౯ࢆᐇ᪋ࡋࡓ㸬ࡇࢀࡲ࡛ࡢ◊✲࡛㸪≉࡟ᢠ⅖⑕స⏝ࡀᙉ࠸ࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗ࡚࠸ࡿࣜࣥ⬡㉁⏬ศࢆ㉳Ⅼ࡜ࡋ㸪ᅛ┦ᢳฟࡸࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮ࢆ⏝࠸࡚ࣜࣥ⬡㉁ࢆࢡࣛࢫࡈ࡜㸪ศᏊ✀ࡈ࡜࡟⢭〇ࡋࡓ㸬⢭〇ࡋࡓࡶࡢࡢ࠺ࡕ㸪᭱ࡶᢠ⅖⑕స⏝ࡀᙉ࠸ࣜࣥ⬡㉁ࡢศ
Ꮚᵓ㐀ࢆ᥎ᐃࡋࡓ⤖ᯝ࡟ࡘ࠸࡚ሗ࿌ࡍࡿ㸬㸰㸬ᐇ㦂᪉ἲ
 ᮦᩱཬࡧヨ⸆ࢫࢧࣅࣀࣜ࡜ࡋ࡚ 2011 ᖺᗘ᭷᫂ᾏ⏘ᯈᾏⱏࢆ⏝࠸ࡓ㸬ヨᩱࢆ㉸㐲ᚰ⢊○ᶵ㸦ZM-1; Retsch 〇㸧࡛⢊○ࡋ㸪⢏Ꮚᚄ 125 Pm ௨ୗࡢᚤ⢊ᮎࢆᚓࡓ㸬ရ㉁ࡢ
ຎ໬ࢆ㜵ࡄࡓࡵ㸪ᚤ⢊ᮎࡣᐦᑒୗ-18Υ࡛ಖ⟶ࡋࡓ㸬ᮏ◊✲࡟࠾ࡅࡿヨ⸆ࡣ௨ୗࡢࡶࡢࢆ⏝࠸ࡓ㸸
penicillin/streptomycin㸦Life Technologies Gibco㸧, ࢘ࢩ ⫾ ඣ ⾑ Ύ (FBS; Thermo Scientific HyClone),
Dulbecco's Modified Eagle Medium (DMEM; ࿴ග⣧⸆ᕤᴗ) , LPS㸦Escherichia coli 0111:B4 ⏤᮶; ࿴ග⣧⸆ᕤᴗ), mouse TNF-D/IL-6 ELISA Kit (Mouse ELISA
Ready-SET- Go!®; Affymetrix eBioscience), Cell
Counting Kit-8 (ྠோ໬Ꮫ◊✲ᡤ), ࢡ࣐ࣟࢺࢢࣛࣇ࢕
࣮⏝ࢩࣜ࢝ࢤࣝ (Wakogel C-300HG; ࿴ග⣧⸆ᕤᴗ),1,2-dipalmitoyl-phosphatidylglycerol(diP-PG),
1palmitoyl-2-arachidonoyl-phosphatidylglycerol (PA-PG),
1-palmitoyl-2-oleoyl-phosphatidylglycerol(PO-PG),
1-palmitoyl-2-oleoyl-phosphatidylethanolamine (PO-PE),
1-palmitoyl-2-oleoyl-phosphatidic acid (PO-PA),
1palmitoyl-2-oleoyl-phosphatidylserine(PO-PS),
1palmitoyl-2-oleoyl-phosphatidylcholine (PO-PC), 1palmitoyl-2-hydroxy-phosphatidylcholine (P-LPC) ( ௨ୖ 8 Ⅼ⥲࡚ Avanti Polar Lipid), 1,2-dieicosapentaenoyl
phosphatidylcholine, methyl palmitoleate,
methylpalmitate, methyl stearate, methyl oleate, methyl
linoleate,
methyleicosapentaenoate,
methylarachidonate (௨ୖ 8 Ⅼ⥲࡚ Sigma-Aldrich), methyl
3-hydroxypalmitate (Matreya LLC), quantitative GC
mixture, GLC-60 (containing 4 C20-series fatty acidmethyl esters, eicosanoate, cis-11- eicosenoate, cis11,14-eicosadienoate, and cis-11,14,17-eicosatrienoate;
Matreya LLC), phospholipase A2 (Naja mossambica
mossambica ⏤ ᮶ ; Sigma-Aldrich) ,  TMS-HT(HMDS/TMCS/pyridine=3/1/9 (v/v/v); ᮾி໬ᡂᕤᴗ).
ࡲࡓ㸪᭷ᶵ⁐፹ཬࡧ㓟࣭࢔ࣝ࢝ࣜヨ⸆ࡣ㸪≉࡟グ㍕ࡋ࡞࠸㝈ࡾ࿴ග⣧⸆ᕤᴗ〇ࡢ≉⣭ヨ⸆, HPLC ⏝ヨ⸆ࡶࡋࡃࡣ LC-MS ⏝ヨ⸆ࢆ⏝࠸ࡓ㸬 ⣽⬊ᇵ㣴ヨ㦂 ࣐࢘ࢫ⏤᮶࣐ࢡࣟࣇ࢓࣮ࢪᵝ⣽⬊ᰴ RAW264.7
㸦⌮◊ࣂ࢖࢜ࣜࢯ࣮ࢫࡼࡾᥦ౪㸧ࢆ⏝࠸࡚⣽⬊ヨ㦂ࢆ⾜ࡗࡓ㸬⣽⬊ࡣ 1% penicillin/streptomycin, 10% FBS
ྵ᭷ DMEM ᇵᆅ࡟ࡼࡾ 5%CO2 Ꮡᅾୗ 37Υ࡛⥅௦ᇵ㣴ࡋࡓ㸬LPS ㄏᑟᆺ NO ཬࡧ TNF-D/IL-6 ࡢ⏘⏕ヨ㦂ࡣ㸪᪤ሗ 7)࡟ᚑࡗࡓ㸬 ヨᩱࡣ⁐፹ࢆ᏶඲࡟␃ཤࡋࡓ
ᚋ㸪ᇵᆅ࡟ᠱ⃮ࡉࡏ㸪㉸㡢Ἴฎ⌮࡟ࡼࡾศᩓࡉࡏࡓ㸬TNF-D⃰ᗘཬࡧ IL-6 ⃰ᗘࡣᕷ㈍࢟ࢵࢺࢆ⏝࠸ࡓ
sandwich ELISA ࡟ࡼࡾồࡵࡓ㸬 NO ⏘⏕㔞ࡣ㸪Griessἲ 11)࡟ࡼࡾᇵ㣴ୖΎ࡟⵳✚ࡋࡓள◪㓟࢖࢜ࣥ㸦NO2-㸧⃰ᗘࢆ ᐃࡍࡿࡇ࡜࡟ࡼࡾ⟬ฟࡋࡓ㸬ࡇࢀࡽࡢ⅖⑕ᛶ≀㉁ࡢ⏘⏕㜼ᐖ⋡ࡣ㸪ᘧ㸦1㸧࡟ࡼࡾ⟬ฟࡋࡓ㸬⏘⏕㜼ᐖ⋡ (%)
= {1–('Aa–'Ab)/('Ac– 'Ab)}×100࣭࣭࣭㸦㸯㸧ࡇࡇ࡛㸪'Aa㸸ヨᩱῧຍᇵᆅࡢ྾ගᗘᕪ
'Ab㸸ࣈࣛࣥࢡࡢ྾ගᗘᕪ
'Ac㸸ࢥࣥࢺ࣮ࣟࣝࡢ྾ගᗘᕪ㸬ᚲせ࡟ᛂࡌ㸪ྛヨᩱῧຍ༊ࡢࢥࣥࢺ࣮ࣟࣝ༊࡟ᑐ
ࡍࡿ⏘⏕㜼ᐖ⋡ࡢᕪࢆ㸪Dunnett ࡢከ㔜ẚ㍑࡟ࡼࡾ᳨ᐃࡋࡓ㸬 ⣽⬊യᐖᛶ⣽⬊ヨ㦂࡟ࡼࡿ⅖⑕ᛶ࣓ࢹ࢕࢚࣮ࢱ࣮⏘⏕ᢚไࡢጇᙜᛶࢆ☜ㄆࡍࡿ࡭ࡃ㸪⣽⬊άᛶ ᐃ࢟ࢵࢺ㸦Cell
Counting Kit-8㸧ࢆ⏝࠸࡚㸪ヨᩱࡢ⣽⬊㞀ᐖᛶࢆ ᐃࡋࡓ㸬2. 2 ࡜ྠ୍ࡢᇵ㣴᮲௳࡛ヨᩱཬࡧ LPS ࡢᏑᅾୗ 20 ᫬㛫⣽⬊ᇵ㣴ࡋ㸪ᇵᆅࢆ୍᪦㝖ཤᚋ㸪Cell
Counting Kit-8 ࢆ ྵ ࡴ ᇵ ᆅ 㸦 2 PL reagent/98 PL
medium㸧࡛ 6 ᫬㛫ᇵ㣴ࡋ㸪࣐࢖ࢡࣟࣉ࣮ࣞࢺ࣮ࣜࢲ
࣮࡟࡚Ἴ㛗 450 nm ࡜ 650 nm ࡟࠾ࡅࡿ྾ගᗘᕪ㸦'A㸧ࢆ ᐃࡋࡓ㸬⣽⬊യᐖᛶࡢ⛬ᗘࡣ㸪ࢥࣥࢺ࣮ࣟࣝ༊
࡟࠾ࡅࡿ'A ࡟ᑐࡍࡿヨᩱῧຍᇵᆅ༊ࡢ'A ࡢẚ࡛⾲ࡋࡓ㸬 ⬡㉁ࡢᢳฟཬࡧศ⏬
Chloroform/methanol ΰྜ⁐፹㸦C/M=1/1,v/v㸧ࢆ⏝࠸࡚㸪ࢫࢧࣅࣀࣜ⢊ᮎࡼࡾ⬡㉁ࢆᢳฟࡋࡓ㸬1 kg ࡢ䋭 㪉㪇 䋭

ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠ

ࢫࢧࣅࣀࣜ⢊ᮎ࡟ 8 L ࡢ⁐፹ࢆῧຍࡋ㸪ᖖ ࡟࡚ 1᫬㛫᧠ᢾࡋࡓ㸬ࡇࢀࢆῶᅽℐ㐣࡟ࡼࡾᅛᾮศ㞳ࡋ㸪ᢳฟᾮࢆᅇ཰ࡋࡓ㸬ᢳฟṧ´࡟෌ᗘ 5 L ࡢ⁐፹ࢆຍ࠼㸪ᖖ ࡟࡚ 1 ᫬㛫᧠ᢾࡋࡓ㸬ῶᅽℐ㐣࡟ࡼࡾᢳฟ
ᾮࢆᚓ㸪➨୍ࡢᢳฟᾮ࡜ྜࢃࡏ࡚ࡇࢀࢆῶᅽ⃰⦰ࡋ㸪
⣙ 1.5 L ࡢࢫࢧࣅࣀࣜ⬡㉁⃰⦰ᾮࢆᚓࡓ㸬ࡇࢀ࡟
chloroform/methanol/0.9% KCl Ỉ⁐ᾮ=1/1/0.9 (v/v/v)
࡟࡞ࡿࡼ࠺ྛ⁐፹ࢆῧຍࡋ㸪᣺┞ᚋ㸪4Υ࡟୍࡚ᬌ㟼⨨ࡋ࡚ 2 ᒙศ㞳ࡉࡏࡓ㸬ୖᒙࢆៅ㔜࡟㝖ཤࡋ㸪ୗᒙࢆῶᅽ⃰⦰ࡋ࡚⁐፹ࢆ␃ཤࡋ㸪49.1 g ࡢࢫࢧࣅࣀࣜ
⏤᮶⬡㉁ࢆᚓࡓ㸬ᢳฟࡋࡓ⬡㉁ࡣⰍ⣲ࡸ༢⣧⬡㉁ࢆྵࡴࡓࡵ㸪ࢩࣜ࢝ࢤ࣒ࣝ࢝ࣛࢆ⏝࠸ࡓᅛ┦ᢳฟἲ 12)࡟ࡼࡾ」ྜ⬡㉁ࢆศ⏬ࡋࡓ㸬Chloroform ࡛ᖹ⾮໬ࡋࡓࢩࣜ࢝ࢤࣝ࢝
࣒ࣛ㸦ࢤࣝᐜ㔞 240 mL㸪࢞ࣛࢫ〇ࢡ࣐ࣟࢺ⟶㸦i.d.60
x length 500 mm㸧࡟ࣃࢵ࢟ࣥࢢ㸧࡟㸪chloroform ࡟⁐ゎࡋࡓ⬡㉁ 5 g ࢆ࢔ࣉࣛ࢖ࡋࡓ㸬࣒࢝ࣛᐜ㔞ࡢ 7
ಸ㔞ࡢ chloroform ࢆῧຍࡋ㸪chloroform ⁐ฟ⏬ศࢆᚓࡓ㸬ḟ࡟㸪࣒࢝ࣛᐜ㔞ࡢ㸵ಸ㔞ࡢ acetone ࢆ࢔ࣉࣛ࢖ࡋ㸪acetone ⁐ฟ⏬ศࢆᚓࡓ㸬᭱ᚋ࡟㸪࣒࢝ࣛᐜ㔞ࡢ 5 ಸ㔞ࡢ methanol ࢆ࢔ࣉࣛ࢖ࡋ㸪methanol ⁐ฟ⏬ศࢆᚓࡓ㸬
ࣜࣥ⬡㉁ࢆከࡃྵࡴ methanol ⁐ฟ⏬ศ 7) ࢆ㸪acetone/methanol㸦A/M㸧ΰྜ⁐፹ࢆ⏝࠸ࡓᅛ┦ᢳฟἲ࡟ࡼࡾࡉࡽ࡟ศ⏬ࡋࡓ㸬Chloroform ࡟⁐ゎࡋࡓ
methanol ⁐ฟ⏬ศࢆ㸪acetone ࡛ᖹ⾮໬ࡋࡓࢩࣜ࢝ࢤ
࣒ࣝ࢝ࣛ㸦ᐜ㔞㸸21.7 mL㸧࡟࢔ࣉࣛ࢖ࡋ㸪acetone, A/M= 9/1, A/M = 8/2, A/M = 7/3, ཬࡧ methanol ࡛㡰ḟ⁐ฟࡉࡏ㸦125 mL ࡎࡘ㸧㸪ྛࠎࡢ⏬ศࡢ⁐፹ࢆ␃ཤࡋ
࡚⣽⬊ヨ㦂࡟౪ࡋࡓ㸬 ᢠ⅖⑕⬡㉁ࡢ⢭〇⣽⬊ヨ㦂ࡢ⤖ᯝ㸪ᢠ⅖⑕స⏝ࡀ᭱ࡶᙉ࠿ࡗࡓ A/M= 8/2 ⁐ฟ⏬ศࢆ㸪Diol ࣒࢝ࣛࢆ⏝࠸ࡓ㡰┦㧗㏿ᾮయࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮㸦NP-HPLC㸧࡟ࡼࡾࡉࡽ࡟ศ⏬
ࡋࡓ㸬ࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮ࡢ᮲௳ࡣୗグࡢ࡜࠾ࡾ࡛࠶ࡿ㸸⿦⨨, ᪥ᮏศᯒᕤᴗ〇 NEXT; ࣒࢝ࣛ, Develosil
100 Diol-5 (i.d. 8 x length 250 mm, 㔝ᮧ໬Ꮫ); ⛣ື┦,
hexane/2-propanol/acetic acid/1 mmol/L ammonium
acetate = 68/20/0.07/3 (v/v/v/v); ὶ㏿, 3 mL/min; ᳨ฟ,UV at 214 nm; ὀධ㔞, 7.5 mg/150 PL; ศ⏬ࢫࢣࢪ࣮ࣗࣝ, ὀධ 3 ศᚋࡼࡾ 1 ศ㛫㝸࡛ 9 ᮏศྲྀ㸪ࡑࡢᚋ
12㹼49.5 ศࡲ࡛ 1.5 mL 㛫㝸࡛ 24 ᮏศྲྀ㸬ࢡ࣐ࣟࢺࢢ࣒ࣛ࡞ࡽࡧ࡟ྛࣇࣛࢡࢩࣙࣥࡢ TLC
㸦ࢹ࣮ࢱࡣ♧ࡉࡎ㸧࡟ࡼࡿࣂࣥࢻ఩⨨ࡼࡾ㸪ࣇࣛࢡࢩࣙࣥ A ࠿ࡽ H ࡲ࡛ࡢ 8 ࡘࡢ⏬ศ࡟ࡲ࡜ࡵࡓ㸬ྛࣇࣛࢡࢩࣙࣥࡣ㸪⁐፹ࢆ␃ཤᚋ࡟ 2 mL ࡢ C/M =1/1 ࡟෌⁐ゎࡉࡏ㸪ḟ㡯࡟♧ࡍᡭἲ࡟ࡼࡾ⬡⫫㓟ྵ㔞ࢆᐃࡋࡓ㸬⬡⫫㓟ྵ㔞ࡼࡾྛࣇࣛࢡࢩࣙࣥࡢࣔࣝ⃰ᗘࢆ᥎⟬ࡋ㸪⤊⃰ᗘ 10, 20 Pmol/L ࡛ RAW264.7 ⣽⬊࡟ῧຍࡋ࡚⣽⬊ヨ㦂ࢆ⾜ࡗࡓ㸬 ⬡㉁ࡢᐃᛶ࣭ᐃ㔞ⷧᒙࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮㸦TLC㸧࡟ࡼࡾ⬡㉁ࡢศ
⏬≧ែࢆ☜ㄆࡋࡓ㸬ᒎ㛤᮲௳ࡣ௨ୗࡢ㏻ࡾ࡛࠶ࡿ㸸ࣉ࣮ࣞࢺ, Silica gel-60 TLC aluminum plates (Merck
Germany);ヨᩱሬᕸ㔞, 150 Pg; ᒎ㛤⁐፹, chloroform/methanol/water/triethylamine = 30/35/7/35 (v/v/v/v);ⓎⰍ, Dittmer-Lester ヨ⸆㸬ศ⏬≀ࡢ⬡㉁ྵ㔞ࡣ㸪⬡⫫㓟ྵ㔞ࢆ࢞ࢫࢡ࣐ࣟࢺ
ࢢࣛࣇ࢕࣮-㉁㔞ศᯒ⿦⨨㸦GC-MS㸧࡟ࡼࡾᐃ㔞ࡍࡿࡇ࡜࡛᥎⟬ࡋࡓ㸬ヨᩱࡣ㸪0.5 mol/L Ỉ㓟໬ࢼࢺࣜ࢘
࣒-methanol ฎ⌮࡟⥆ࡃ BF3-methanol ⁐ᾮฎ⌮࡛࣓ࢳ࢚ࣝࢫࢸࣝ໬ࡋࡓᚋ㸪ࢩࣜࣝ໬๣ 㸦TMS-HT)࡛
TMS ໬ࡋ࡚ GC-MS ࡟ᑟධࡋࡓ㸬GC-MS ࡢ᮲௳ࡣୗ
グࡢ࡜࠾ࡾ࡛࠶ࡿ㸸⿦⨨, ᓥὠᰴᘧ఍♫〇 GC2010 + QP2010; ࣒࢝ࣛ,Sigma-Aldrich Co 〇 SP2380㸦length 100 m x i.d. 0.25mm x film thickness 0.2 Pm㸧; ࢟ࣕࣜ࢔࢞ࢫὶ㏿, 20
cm/sec㸦helium㸧; ࣒࢝ࣛ ᗘ, 140Υ for 5 min,140-205 Υ at 1.5 Υ /min, 205-240 Υ at 10 Υ /min,240Υ for 8 min; ὀධ, 1 PL; ࢫࣉࣜࢵࢺẚ 20:1 at
250Υ; ᳨ฟ, EI-MS㸬
 ᵓ㐀ゎᯒ
NP-HPLC ࡟ࡼࡿศ⏬≀ࡢ࠺ࡕ㸪ᢠ⅖⑕స⏝ࡀ᭱ࡶᙉ࠸⏬ศࢆ C/M =1/1 ࡟෌⁐ゎࡉࡏ㸪ࡉࡽ࡟ methanol࡛ 10 ಸᕼ㔘ࡋ࡚⃰ᗘ 100 Pg/mL ࡜ࡋ㸪࢚ࣞࢡࢺࣟࢫ
ࣉ࣮ࣞ࢖࢜ࣥ໬-࢖࢜ࣥࢺࣛࢵࣉᆺ㉁㔞ศᯒィ
㸦ESI-ITMS㸧࡟ᑟධࡋࡓ㸬ࡲࡓ㸪ࢢࣜࢭ࣮ࣟࣝ㦵᱁ࡢ 2 ఩࡟࢚ࢫࢸࣝ⤖ྜࡍࡿ⬡⫫㓟ࢆ≉␗ⓗ࡟ຍỈศゎࡍࡿ㓝⣲ phospholipase
A2 㸦PLA2㸧ࢆ⏝࠸࡚㸪ᵓᡂ⬡⫫㓟ࡢ⤖ྜ఩⨨ࢆ≉ᐃࡋࡓ㸬ศ⏬≀ 50 Pg ࢆ diethylether/methanol = 98/2
(v/v) 1 mL ࡟⁐ゎࡉࡏ㸪Naja mossambica mossambica⏤᮶ PLA2 ࢆ 200 mU ῧຍࡋ㸪ᖖ ࡛ 3 ᫬㛫᧠ᢾࡋࡓ㸬཯ᛂᾮࢆ㐲ᚰศ㞳㸦1,000 x g , 10 min㸧ࡋ࡚᭷ᶵ⁐፹ᒙࢆᅇ཰ࡋ㸪㐲ᚰ⃰⦰࡟ࡼࡾ⁐፹ࢆ␃ཤࡋࡓ㸬஝ᅛ≀࡟ methanol 0.2 mL ࢆῧຍࡋ㸪ESI-ITMS ࡟౪ࡋࡓ㸬ESI-ITMS ࡢ᮲௳ࡣୗグࡢ࡜࠾ࡾ࡛࠶ࡿ㸸⿦⨨, Bruker Daltonics 〇 HCT Ultra; ࢖࢜ࣥ※, ESI;࣏ࣛࣜࢸ࢕, ㈇࢖࢜ࣥ; ὶ㔞, 3 PL/min; ࢖࢜ࣥ※ᗘ, 300Υ; ࢿࣈࣛ࢖ࢨ࣮࢞ࢫᅽຊ, 10 psi; ࢿࣈࣛ࢖ࢨ࣮࢞ࢫὶ㔞, 5 L/min; ࢟ࣕࣆ࣮ࣛࣜ㟁఩, 4 kV; end䋭 㪉㪈 䋭

ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠ

plate offset, 0.5 kV; ࢫ࢟ࣕࣥ⠊ᅖ, 50-1500 m/z; ࢫ࢟ࣕࣥࢫࣆ࣮ࢻ, ẖ⛊ 8100 m/z㸬㸱㸬⤖ᯝศ⏬≀ࡢάᛶẚ㍑
 ࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁ࢆ A/M ΰྜ⁐፹ࢆ⏝࠸ࡓ SPE ࡟ࡼࡾ A, A/M = 9/1, A/M = 8/2, A/M = 7/3, ཬࡧ M ⁐ฟ⏬ศ࡟ 5 ศ⏬ࡋࡓ㸬ྛ⏬ศࡢ཰⋡ࢆ⾲ 1࡟㸪άᛶ໬࣐ࢡࣟࣇ࢓࣮ࢪࡢ⅖⑕ᛶ࣓ࢹ࢕࢚࣮ࢱ࣮
⏘⏕࡟ཬࡰࡍᙳ㡪ࢆ⾲ 2, 3 ཬࡧ 4 ࡟♧ࡍ㸬཰⋡ࡀ᭱ࡶ㧗࠿ࡗࡓࡢࡣ AM = 8/2 ⁐ฟ⏬ศ࡛࠶ࡗࡓ㸬ࡲࡓ㸪ྛῧຍ⃰ᗘ࡟࠾ࡅࡿ NO ⏘⏕㜼ᐖ㸪TNF-D⏘⏕㜼ᐖ㸪⾲㸯 ࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁ࡢSPE࡟࠾ࡅࡿ཰⋡Yield 1
PowderYield 2
wt%

Yield 3

Lipid class, idetified

100.0

Crude lipid

4.9

100.0

1.4

27.8

1st SPE
Methanol eluent

100.0

PLs, GLs

2nd SPE
Acetone

0.24

4.9

17.7

GLs

A/M=9/1

0.22

4.4

15.8

GLs, PG

A/M=8/2

0.42

8.5

30.6

PG, PE

A/M=7/3

0.12

2.4

8.5

PE

Methanol

0.22

4.4

16.0

PC, LPC

Yield 1, ࣀࣜ⢊ᮎࢆ100࡜ࡋࡓ࡜ࡁࡢ཰⋡; Yield 2, ⢒⬡㉁ࢆ100࡜ࡋࡓ࡜ࡁࡢ཰⋡; Yield 3, ࣓ࢱࣀ࣮ࣝ⁐ฟ⏬ศࢆ100࡜ࡋࡓ࡜ࡁࡢ཰⋡; PL, ࣜࣥ⬡㉁; GL, ⢾⬡㉁; PG, phosphatidylglycerol; PE,
phosphatidylethanolamine; PC, phosphatidylcholine; LPC, lysophosphatidylcholine⾲㸰ࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁ࡢSPE⏬ศ࡟࠾ࡅࡿᢠ⅖⑕స⏝㸦NO⏘⏕㜼ᐖ㸧ࡢẚ㍑

ヨᩱ⃰ᗘ(Pg/mL)(Pg/mL)NO
NO⏘⏕㜼ᐖ⋡⏘⏕㜼ᐖ⋡ (%)
Acetone

AM=9/1

A/M=8/2

A/M=7/3

Methanol

10.0

67.4 ±

2.5

69.7 ±

5.4

89.0 ±

0.2

59.4 ±

4.0

11.6 ±

1.0

21.8 ±

5.8

34.6 ±

1.3

33.5 ±

3.8

2.8 ±

2.8

-11.8 ± 24.0

0.1

12.8 ±

7.0

7.8 ±

6.1

16.3 ±

4.3-13.1 ±

5.5

-11.6 ± 13.4

⾲㸱

3.9

ࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁ࡢSPE⏬ศ࡟࠾ࡅࡿᢠ⅖⑕స⏝㸦TNF-D⏘⏕㜼ᐖ㸧ࡢẚ㍑

ヨᩱ⃰ᗘ(Pg/mL)(Pg/mL)TNF-D⏘⏕㜼ᐖ⋡ (%)
TNF-D⏘⏕㜼ᐖ⋡

Acetone

AM=9/1

A/M=8/2

A/M=7/3

Methanol

10.0

22.2 ±

1.8

27.1 ±

4.1

35.8 ±

6.2

18.1 ±

4.0

8.4 ±

1.0

5.1 ±

3.2

15.9 ±

8.9

17.0 ±

9.7

0.5 ±

6.0

18.4 ± 10.1

0.1

-13.9 ±

6.8

-2.4 ±

2.4

0.2 ±

4.3

7.6 ±

2.3

-2.2 ±

⾲㸲

4.7

5.8

ࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁ࡢSPE⏬ศ࡟࠾ࡅࡿᢠ⅖⑕స⏝㸦IL-6⏘⏕㜼ᐖ㸧ࡢẚ㍑

ヨᩱ⃰ᗘ(Pg/mL)(Pg/mL)IL-6 ⏘⏕㜼ᐖ⋡ (%)
Acetone

AM=9/1

A/M=8/2

50.7 ±

4.8

Methanol

10.0

77.5 ±

2.2

80.4 ±

3.3

94.9 ±

1.0

38.5 ±

8.8

38.1 ±

6.5

48.5 ± 10.2

-4.3 ± 12.2

28.6 ± 10.6

0.1

10.4 ± 10.1

-2.5 ± 14.7

16.1 ± 10.8

-0.6 ±-1.2 ±

䋭 㪉㪉 䋭

1.9

A/M=7/3

4.3

43.0 ±

6.8

8.3

ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠ

IL-6 ⏘⏕㜼ᐖ࠸ࡎࢀ࡟࠾࠸࡚ࡶ㸪᭱ࡶ㧗࠸㜼ᐖ⋡ࢆ♧ࡋࡓࡢࡣ㸪A/M = 8/2 ⁐ฟ⏬ศ࡛࠶ࡗࡓ㸬TLC ࡢ⤖ᯝ㸪A/M = 8/2 ⁐ฟ⏬ศ࡟࠾࠸᳨࡚ฟࡉࢀࡿ୺せ࡞ࣂࣥࢻࡢ Rf ್ࡣ㸪phospatidylglucerol (PG) ࡜࡯ࡰྠ୍࡛࠶ࡾ㸪ࡇࡢ⏬ศࡀ PG ࢆ୺ᡂศ࡜ࡋ࡚ྵࡴࣜࣥ⬡㉁࡛࠶ࡿࡇ࡜ࢆᙉࡃ♧၀ࡋ࡚࠸ࡓ㸦ᅗ 1㸧㸬࡞࠾㸪
ヨ㦂ࡋࡓ඲࡚ࡢ༊࡟࠾࠸࡚㸪Cell Counting Kit-8 ࡟ࡼࡿ⣽⬊യᐖᛶࡣㄆࡵࡽࢀ࡞࠿ࡗࡓ㸬ḟ࡟㸪A/M = 8/2 ⁐ฟ⏬ศࢆ Diol ࣒࢝ࣛ࡟ࡼࡿ㡰┦ࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮࡟ࡼࡾ⣽ศ⏬ࡋ㸪8 ࡘࡢ⏬ศ䋭 㪉㪊 䋭

ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠ㸦ࣇࣛࢡࢩࣙࣥ A㹼H㸧ࢆᚓࡓ㸦ᅗ 2㸧㸬ࣇࣛࢡࢩࣙ
ࣥ A ࡟ࡘ࠸࡚ࡣ㸪GC-MS ࡟ࡼࡿ⬡⫫㓟⤌ᡂゎᯒ࡟ࡼࡾ࡯࡜ࢇ࡝⬡㉁ࡀྵࡲࢀ࡚࠸࡞࠸ࡇ࡜ࡀุ᫂ࡋࡓ
ࡓࡵ㸪௨ᚋࡢホ౯࠿ࡽ㝖እࡋࡓ㸬ࣇࣛࢡࢩࣙࣥ B㹼
H ࡢ NO, TNF-D, IL-6 ⏘⏕ᢚไస⏝ࢆᅗ 3 ࡟♧ࡋࡓ㸬ྛ⏬ศࡢᢠ⅖⑕స⏝ࢆẚ㍑ࡋࡓ⤖ᯝ㸪ࣇࣛࢡࢩࣙࣥ
E ཬࡧ F ࡟ᙉ࠸ NO ⏘⏕ᢚไస⏝ࡀㄆࡵࡽࢀ㸪࡞࠿

Frc H

A

Frc G
Frc F
Frc E
Frc D

10 nmol/mL

Frc C

20 nmol/mL

Frc B
Initital
0

20

40
60
80
NO Inhibition 䠄 % 䠅

100

Frc H

120

B

Frc G
Frc F
Frc E
Frc D

10 nmol/mL

Frc C

20 nmol/mL

Frc B
Initital
0

20

40
60
80
TNF-D Inhibition 䠄 % 䠅100࡛ࡶࣇࣛࢡࢩࣙࣥ F ࡀ᭱ࡶస⏝ࡀᙉ࠿ࡗࡓ㸬ࡲࡓ㸪ࣇࣛࢡࢩࣙࣥ E ཬࡧ F ࡣ TNF-D, IL-6 ࡢ⏘⏕ᢚไస⏝ࡶᙉࡃ㸪ࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁࡟ྵࡲࢀࡿᢠ⅖⑕⬡㉁ࡢ୺యࡣ㸪ࡇࡢ⏬ศ࡟ศ㓄ࡉࢀ࡚࠸ࡿࡶࡢ࡜
ุ᩿ࡉࢀࡓ㸬࡞࠾㸪ヨ㦂ࡋࡓ඲࡚ࡢ༊࡟࠾࠸࡚㸪Cell
Counting Kit-8 ࡟ࡼࡿ⣽⬊യᐖᛶࡣㄆࡵࡽࢀ࡞࠿ࡗ
ࡓ㸬ᢠ⅖⑕⬡㉁ࡢᵓ㐀ゎᯒࣇࣛࢡࢩࣙࣥ F ࢆ C/M =1/1 ࡟෌⁐ゎࡉࡏ㸪
ESI-ITMS ࡟ᑟධࡋࡓ㸬ࡑࡢ⤖ᯝ㸪ᅗ 4 ࡟♧ࡍࡼ࠺࡟㸪୺せ࡞⬺ࣉࣟࢺࣥศᏊ [M-H] ࡛࠶ࡿ m/z 783.4 ࡞ࡽࡧ࡟ࡑࢀࡽࡢྠ఩యࣆ࣮ࢡࡀ᳨ฟࡉࢀࡓ㸬ࡲࡓ㸪m/z
783.4 ࢆࣉ࣮ࣞ࢝ࢧ࣮࡜ࡍࡿ MS/MS ࡟ࡼࡾ᳨ฟࡉࢀࡿ୺せ࡞ࣇࣛࢢ࣓ࣥࢺࡣ㸪m/z 271.0 ୪ࡧ࡟ m/z 529.1
࡛࠶ࡗࡓ㸬ࡉࡽ࡟㸪MS/MS ࣇࣛࢢ࣓ࣥࢺ m/z 529.1
ࢆࣉ࣮ࣞ࢝ࢧ࣮࡜ࡍࡿ MS3 ࡟ࡼࡾ᳨ฟࡉࢀࡿ୺せ࡞ࣇࣛࢢ࣓ࣥࢺࡣ m/z 257.0 ୪ࡧ࡟ m/z 301.0 ࡛࠶ࡗࡓ㸬௚᪉㸪⾲㸳ࡢ⬡⫫㓟⤌ᡂ࡟♧ࡍࡼ࠺࡟㸪ࣇࣛࢡࢩࣙࣥ F ࡣ㸪࡯ࡰᙜࣔࣝẚࡢ EPA ࡜ 3-hydroxypalmitic
acid (3-OH-PA)࡛ᵓᡂࡉࢀ࡚࠸ࡓ㸬 ࣇࣛࢡࢩࣙࣥ F ࡢ PLA2 ᾘ໬≀ࡢ MS ࢆᅗ 5 ࡟♧ࡍ㸬୺せ࡞⬺ࣉࣟࢺࣥศᏊ [M-H] ࡜ࡋ࡚㸪m/z 271.0࡞ࡽࡧ࡟ m/z 529.1 ࡀほᐹࡉࢀࡓ㸬 ௨ୖࡢ⤖ᯝࡼࡾ㸪ࣇࣛࢡࢩࣙࣥ F ࡣ㸪EPA ࡜
3-OH-PA ࢆᵓᡂ⬡⫫㓟࡜ࡋ㸪sn-2 ఩࡟㉁㔞 272.0 ࡢ⬡⫫㓟ࡀ⤖ྜࡍࡿ㸪ࣔࣀ࢔࢖ࢯࢺࣆࢵࢡ㉁㔞 784.4
ࡢࣜࣥ⬡㉁࡛࠶ࡿ࡜ุ᩿ࡉࢀࡿ㸬3-OH-PA ࡢࣔࣀ࢔࢖ࢯࢺࣆࢵࢡ㉁㔞ࡀ 272.2 ࡛࠶ࡿࡇ࡜࠿ࡽ㸪ࡇࡢࣜࣥ⬡㉁ࡢ sn-2 ఩࡟⤖ྜࡍࡿࡢࡣ 3-OH-PA ࡛࠶ࡿ㸬120

⾲㸳 ࣇࣛࢡࢩࣙࣥEཬࡧࣇࣛࢡࢩࣙࣥF࡟࠾ࡅࡿ
⬡⫫㓟⤌ᡂ

Frc H

C

Frc G

⬡⫫㓟⤌ᡂ 䠄mol%䠅
䝣䝷䜽䝅䝵䞁 E 䝣䝷䜽䝅䝵䞁 F

Frc F

Palmitate (C16:0)

29.2

1.6

trans -13-Hexadecanoate (C16:1n13t)

19.4

1.1

10 nmol/mL

3-OH-Palmitate (3-OH-C16:0)

N.D.

47.1

20 nmol/mL

Stearate (C18:0)

Tr.

N.D.

Oleate (C18:1n9c)

2.6

0.4

Linoleate (C18:2n6c)

Tr.

N.D.

Arachidate (C20:0)

N.D.

N.D.

cis -11-Eicosenoate (C20:1n9c)

N.D.

N.D.

cis -11,14-Eicosadienoate (C20:2n6c)

0.1

N.D.

ᅗ㸱 ࣐ࢡࣟࣇ࢓࣮ࢪࡢLPSㄏᑟᆺNOཬࡧ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥ⏘⏕࡟ཬࡰࡍࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁ศ⏬≀ࡢᙳ㡪cis -8,11,14-Eicosatrienoate (C20:3n6c)

N.D.

N.D.

Arachidonate (C20:4n6c)

0.1

N.D.

EPA (C20:5n3c)

48.5

49.8

A, NO; B, TNF-D; C, IL-6

Total

100.0

100.0

Frc E
Frc D
Frc C
Frc B
Initital
0

20

40
60
80
IL-6 Inhibition 䠄 % 䠅

100

120

䋭 㪉㪋 䋭

ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠ

ࡲࡓ㸪EPA ࡜ 3-hydroxypalmitic acid (3-OH-PA)࡛ᵓᡂ
ࡉࢀࡿ PG ࡢࣔࣀ࢔࢖ࢯࢺࣆࢵࢡ㉁㔞ࡀ㸪⌮ㄽୖ
784.4 ࡛࠶ࡾ㸪ᮏ◊✲࡟࠾ࡅࡿ TLC ୪ࡧ࡟ MS ࡢᐃ⤖ᯝ࡜ྜ⮴ࡍࡿ㸬௨ୖࡢ ᐃ⤖ᯝࡼࡾ㸪ࡇࡢࣜࣥ⬡㉁ࡣ㸪sn-1 ఩࡟Intens.
x108

EPA ࡀ㸪sn-2 ఩࡟ 3-OH-PA ࡀ࢚ࢫࢸࣝ⤖ྜࡍࡿ PG
࡛ ࠶ ࡿ 1-eicosapentaenoyl-2- (3-hydroxypalmitoyl)-
phosphatidylglycerol (EpPOH-PG; ᅗ 6) ࡜᥎ᐃࡉࢀࡓ㸬
MS/MS ࠾ࡼࡧ MS3 ࡟࠾ࡅࡿ ᐃ⤖ᯝࡶ㸪ࡇࡢࣜࣥ⬡㉁ࡀ EpPOH-PG ࡛࠶ࡿࡇ࡜࡜▩┪ࡋ࡞࠸⤖ᯝ࡛࠶
-MS, 0.1-0.5min #(3-28)

䝣䝷䜽䝅䝵䞁 F - MS

0.8

783.4

0.6
0.4
0.2
0.0
100

200

300

400

500

Intens.
x107䝣䝷䜽䝅䝵䞁 F – MS/MS

0.8

271.0

600

700

800

m/z

-MS2(783.4), 0.1-0.5min #(2-15)

0.6
529.1

0.4
0.2

765.3

0.0
100
Intens.
x105

200

300

400

500

600

700

800

m/z

-MS3(783.4->529.1), 0.1-0.5min #(2-12)䝣䝷䜽䝅䝵䞁 F – MS3
301.0

2
257.0

1
152.8

0
100

437.0

200

300

400

500

600

700

800

m/z

ᅗ㸲 ࣇࣛࢡࢩࣙࣥFࡢESI-ITMS࡟ࡼࡿ㉁㔞ศᯒ࠾ࡼࡧࣇࣛࢢ࣓ࣥࢺゎᯒ
Intens.
x107
6-MS, 0.0-1.0min #(2-57)

A

783.4

䝣䝷䜽䝅䝵䞁F

4
2
0
200
Intens.
x108

B

400

271.0

0.8

600

800

Eicosapentaenoyl-hydroxyl-PG,
deprotonated molecular

0.6

1000

m/z
-MS, 0.0-1.0min #(2-56)

3-Hydroxyhexadecanoic acid,deprotonated molecular䝣䝷䜽䝅䝵䞁F
PLA2ฎ⌮

0.4
529.1

0.2
419.1

0.0
200

400

565.3

600

823.4

800

1000

ᅗ㸳 ࣇࣛࢡࢩࣙࣥFཬࡧPLA2ᾘ໬≀ࡢESI-ITMS࡟ࡼࡿ㉁㔞ศᯒ䋭 㪉㪌 䋭

m/z

ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠ

arachidonoyl phosphatidylglycerol (PA-PG) 㸪 1,2dieicosapentaenoyl phosphatidylcholine (diEp-PC)ཬࡧ
EpPOH-PG ࡛࠶ࡾ㸪≉࡟㸪EpPOH-PG ཬࡧ PA-PG ࡟ᙉ࠸ᢚไస⏝ࡀぢࡽࢀࡓ㸬ࡲࡓ㸪⅖⑕ᛶࢧ࢖ࢺ࢝࢖
ࣥ⏘⏕࡟ཬࡰࡍᙳ㡪ࢆぢࡓ࡜ࡇࢁ㸪⾲㸵࡟♧ࡍࡼ࠺
࡟㸪NO ⏘⏕࡟ཬࡰࡍᙳ㡪࡜ྠᵝ㸪PO-PG, PA-PG,
diEp-PG ཬࡧ EpPOH-PG ࡟᫂☜࡞⏘⏕ᢚไస⏝ࢆㄆࡿ㸬ࣜࣥ⬡㉁࡟࠾ࡅࡿᢠ⅖⑕స⏝ࡢẚ㍑
EpPOH-PG ࢆྵࡴ✀ࠎࡢࣜࣥ⬡㉁࡟࠾ࡅࡿάᛶ໬
࣐ࢡࣟࣇ࢓࣮ࢪࡢ NO ⏘⏕ᢚไస⏝ࢆẚ㍑ࡋࡓ㸦⾲㸴㸧㸬ホ౯ࡋࡓࣜࣥ⬡㉁ࡢ࠺ࡕ㸪᫂☜࡞ NO ⏘⏕ᢚไࡀ ㄆ ࡵ ࡽ ࢀ ࡓ ࡢ ࡣ 㸪 1-palmitoyl-2-oleoylphosphatidylglycerol(PO-PG) 㸪 1-palmitoyl-2-

⾲㸴 ࣐ࢡࣟࣇ࢓࣮ࢪࡢLPSㄏᑟᆺNO⏘⏕࡟ཬࡰࡍ✀ࠎࡢࣜࣥ⬡㉁ࡢᙳ㡪

fatty acids
sn-1
sn-2
sn-3
% NO production inhibition
30 nmol/mL
10 nmol/mL
3 nmol/mL

PO-PG

diP-PG

palmitic acid

palmitic acidoleic acid

palmitic acidphosphoglycerol

phosphoglycerol

phosphoglycerol

59
14
1

±
±
±

4
5
5

-14
-4
2

PO-PC
fatty acids
sn-1
sn-2
sn-3
% NO production inhibition
30 nmol/mL
10 nmol/mL
3 nmol/mL

PA-PG

±
±
±

9
11
5

EpPOH-PG

PO-PApalmitic acideicosapentaenoic acid

palmitic acidarachidonic acid

3-hydroxyhexadecanoic acidoleic acidphosphoglycerol

phosphoric acid

95
43
17

±
±
±

3
5
10

100
90
30

±
±
±

2
3
6

-10
-8
-5

±
±
±

7
9
8

diEp-PC

PO-PS

POPE

P-LPC

palmitic acideicosapentaenoic acid

palmitic acid

palmitic acid

palmitic acidoleic acideicosapentaenoic acidoleic acidoleic acid-phosphocholine

phosphocholine

phosphoserine

phosphoethanolamine

phosphocholine

-10
-4
-1

±
±
±

8
8
8

62
29
11

±
±
±

4
4
4

-9
-11
-6

±
±
±

9
8
13

-17
-6
-6

±
±
±

10
8
8

-8
-2
0

±
±
±

13
10
7

⾲㸵 ࣐ࢡࣟࣇ࢓࣮ࢪࡢLPSㄏᑟᆺ⅖⑕ᛶࢧ࢖ࢺ࢝࢖ࣥ⏘⏕࡟ཬࡰࡍ✀ࠎࡢࣜࣥ⬡㉁ࡢᙳ㡪PO-PG% TNF-D production inhibition
30 nmol/mL
10 nmol/mL
3 nmol/mL% IL-6 production inhibition
30 nmol/mL
10 nmol/mL
3 nmol/mL% WST Activity
30 nmol/mL
10 nmol/mL
3 nmol/mL

diP-PG

EpPOH-PG

PO-PA

81
54
20

±
±
±

2
4
2

-26
6
11

±
±
±

18
16
6

97
52
24

±
±
±

2
13
21

105
88
58

±
±
±

1
6
7

-1
-7
0

±
±
±

2
18
7

99
87
64

±
±
±

0
2
3

13
22
7

±
±
±

4
4
27

94
84
38

±
±
±

2
7
5

92
97
81

±
±
±

3
2
7

2
-3
-2

±
±
±

2
2
20

103
102
98

±
±
±

1
2
4

98
99
98

±
±
±

1
4
4

104
103
104

±
±
±

2
1
1

105
105
104

±
±
±

2
2
2

89
90
99

±
±
±

12
7
3

PO-PC
% TNF-D production inhibition
30 nmol/mL
10 nmol/mL
3 nmol/mL% IL-6 production inhibition
30 nmol/mL
10 nmol/mL
3 nmol/mL% WST Activity
30 nmol/mL
10 nmol/mL
3 nmol/mL

PA-PG

diEp-PC

PO-PS

POPE

P-LPC

2
3
-5

±
±
±

6
13
1

48
26
8

±
±
±

9
7
12

-5
1
9

±
±
±

2
11
9

19
11
1

±
±
±

12
14
8

-11
-5
0

±
±
±

14
5
8

4
-6
1

±
±
±

39
15
12

71
36
-9

±
±
±

5
1
36

-11
-14
13

±
±
±

12
29
6

25
18
15

±
±
±

15
15
7

1
2
17

±
±
±

15
9
7

97
96
98

±
±
±

4
2
1

103
101
99

±
±
±

1
0
1

95
88
88

±
±
±

2
4
13

84
98
94

±
±
±

8
2
3

104
102
99

±
±
±

0
2
2

䋭 㪉㪍 䋭

ᐔᚑ 25 ᐕᐲ૒⾐⋵Ꮏᬺᛛⴚ࠮ࡦ࠲࡯⎇ⓥႎ๔ᦠ

OH

OH
O

P

OH
O

O
O
O
O

O

OH

ᅗ6 ᪂つᢠ⅖⑕ᛶࣜࣥ⬡㉁1-eicosapentaenoyl-2(3-hydroxypalmitoyl)-phosphatidylglycerol (EpPOHPG)ࡢศᏊᵓ㐀᥎ᐃࡵ㸪≉࡟ EpPOH-PG ࡛ࡑࡢస⏝ࡀᙉ࠿ࡗࡓ㸬ヨ㦂ࡋࡓ඲࡚ࡢ༊࡟࠾࠸࡚㸪Cell Counting Kit-8 ࡟ࡼࡿ⣽⬊യᐖᛶ㸦WST άᛶ㸧ࡣㄆࡵࡽࢀ࡞࠿ࡗࡓ㸬㸲㸬⪃ᐹᡃࠎࡣ㸪in vitro ⅖⑕ࣔࢹࣝ࡜ࡋ࡚ከ⏝ࡉࢀ࡚࠸ࡿ
࣐࢘ࢫ࣐ࢡࣟࣇ࢓࣮ࢪᵝ⣽⬊ᰴ RAW264.7 ࢆ⏝࠸㸪ࢫࢧࣅࣀࣜ⏤᮶ᢠ⅖⑕ᛶࣜࣥ⬡㉁ࡢ≉ᐃ໬ࢆヨࡳࡓ㸬ᢠ⅖⑕స⏝ࡢᣦᶆ࡜ࡋ࡚㸪LPS ࡟ࡼࡾㄏᑟࡉࢀࡿ 3
ࡘࡢ⅖⑕ᛶ࣓ࢹ࢕࢚࣮ࢱ㸦NO, TNF-D, IL-6㸧ࡢ⏘⏕ᢚไ⋡࡛ẚ㍑ࡋࡓ㸬ࢫࢧࣅࣀࣜ⏤᮶ࣜࣥ⬡㉁࠿ࡽ㸪SPE ཬࡧ HPLC ࡟ࡼࡾᵓ㐀ࡢ␗࡞ࡿ⬡㉁ࢆศ⏬ࡋ㸪ྛࠎࡢᢠ⅖⑕స⏝ࢆẚ㍑ࡋࡓ㸬ࡑࡢ⤖ᯝ㸪EPA ࡜
3-OH-PA ࡜࡛ᵓᡂࡉࢀࡿ PG ࡀ㸪ᙉ࠸ᢠ⅖⑕స⏝ࢆ᭷ࡍࡿࡇ࡜ࢆぢฟࡋࡓ㸬PLA2 ᾘ໬≀ࡢ MS ほᐹ⤖ᯝࡼࡾ㸪ࡇࡢ⬡㉁ࡢ sn-2 ఩࡟࢚ࢫࢸࣝ⤖ྜࡍࡿ⬡⫫㓟ࡣ 3-hydroxypalmitic acid ࡛࠶ࡿࡇ࡜ࢆ≉ᐃࡋࡓ㸬௨ୖࡢᐇ㦂⤖ᯝࡼࡾ㸪ࡇࡢᢠ⅖⑕ᛶࣜࣥ⬡㉁ࡣ㸪
1-eicosapentaenoyl-2-(3-hydroxypalmitoyl)- phosphatidylglycerol (EpPOH- PG) ࡛࠶ࡿ࡜᥎ᐃࡋࡓ㸦ᅗ 6㸧㸬
EpPOH-PG ࢆྵࡴ࠸ࡃࡘ࠿ࡢࣜࣥ⬡㉁࡟࠾ࡅࡿᢠ
⅖⑕స⏝ࢆẚ㍑ࡋࡓ࡜ࡇࢁ㸪⬡㉁ࢡࣛࢫࡢ୰࡛ࡶ㸪≉࡟ PG ࡟ᙉ࠸స⏝ࡀㄆࡵࡽࢀࡓࡀ㸪ࡑࡢస⏝ࡢ⛬ᗘࡣ㸪⤖ྜࡍࡿ⬡⫫㓟࡟ࡼࡗ࡚␗࡞ࡾ㸪ㄪᰝࡋࡓ⠊ᅖ࡛ࡣ㸪άᛶࡢᙉࡉࡣ EpPOH-PG > PA-PG > PO-PG
࡛㸪diP-PG ࡟ࡣ඲ࡃάᛶࡀぢࡽࢀ࡞࠿ࡗࡓ㸬ࡲࡓ㸪
phosphatidylcholine (PC) ࡟࠾࠸࡚ࡣ㸪PO-PC ࡟ࡣ඲ࡃస⏝ࡀㄆࡵࡽࢀ࡞࠿ࡗࡓࡀ㸪EPA ࡀ 2 ࡘ⤖ྜࡍࡿ
diEp-PC ࡟ࡣᢠ⅖⑕స⏝ࡀㄆࡵࡽࢀࡓ㸬ࡇࢀࡽࡢࡇ
࡜࠿ࡽ㸪άᛶ໬ࡋࡓ࣐ࢡࣟࣇ࢓࣮ࢪ࡟ᑐࡍࡿࣜࣥ⬡㉁ࡢᢠ⅖⑕స⏝ࡢᙉࡉࡣ㸪⤖ྜࡍࡿ⬡⫫㓟㸦ᑿ㒊㸧࡜ࣜࣥ㓟࢚ࢫࢸࣝࢆྵࡴ㢌㒊ࡢ✀㢮࡟ࡼࡾ␗࡞ࡿࡇ
࡜ࡀ♧၀ࡉࢀࡿ㸬ᚓࡽࢀࡓᐇ㦂⤖ᯝ࠿ࡽᢠ⅖⑕స⏝࡜ศᏊᵓ㐀ࡢ㛵ಀᛶࢆぢࡿ㝈ࡾ㸪ᑿ㒊ࡢ⬡⫫㓟ࡢ୙㣬࿴ᗘࡀ㧗࠸ࡇ࡜࡜㸪㢌㒊ࡀ phosphoglycerol ࡛࠶ࡿࡇ࡜ࡢ㸰Ⅼࡀ㸪ࣜࣥ⬡㉁࡟࠾ࡅࡿᢠ⅖⑕స⏝ࡢⓎ⌧࡟࡜ࡗ࡚㔜せ࡛࠶ࡿ࡜⪃࠼ࡽࢀࡓ㸬⫵⬊ࢧ࣮ࣇ࢓ࢡࢱࣥࢺ⏤᮶ࣜࣥ⬡㉁ࡢᢠ⅖⑕స⏝࡟㛵ࡍࡿ Kuronuma ࡽࡢ◊✲ 13)࡟ࡼࢀࡤ㸪PO-PG ࡸphosphatidylinositol ࡞࡝ࡢ㓟ᛶࣜࣥ⬡㉁ࡣ㸪ࣄࢺࣜࣥࣃ⭘⏤᮶⣽⬊ᰴ U937 ࡢ TLR-4 ࢆ௓ࡋࡓࢧ࢖ࢺ࢝࢖
ࣥ⏘⏕ࢆᢚ࠼ࡿࡇ࡜ࡀ᫂ࡽ࠿࡜࡞ࡗ࡚࠸ࡿ㸬ࡲࡓ㸪
Kandasamy14)ࡽࡣ㸪Mycoplasma pneumoniae ࡢ่⃭࡟ࡼࡿ TLR-2 ࢆ௓ࡋࡓ RAW264.7 ⣽⬊ࡢ࢚࢖ࢥࢧࣀ࢖
ࢻ⏘⏕ࢆ㸪PO-PG ࡀᢚ࠼ࡿࡇ࡜ࢆ᫂ࡽ࠿࡟ࡋ࡚࠸ࡿ㸬ࡇࡢࡼ࠺࡟㸪PO-PG ࡟࠾ࡅࡿ TLR-4 ࡸ TLR-2 ࡢ࢔ࣥࢱࢦࢽࢫࢺస⏝ࡀሗ࿌ࡉࢀ㸪ࡑࡢ⸆⌮ⓗຠᯝࡸస⏝ᶵᗎࡢゎ᫂࡟ὀ┠ࡀ㞟ࡲࡗ࡚࠸ࡿ㸬ᮏ◊✲࡟࠾ࡅࡿ EpPOH-PG ࡟ࡶ PO-PG ࡜ྠ➼௨ୖࡢຠᯝࡀᮇᚅ
ࡉࢀ㸪ࡇࡢ⬡㉁ࡢయෆ྾཰ᛶࡸືែ㸪࢔ࣥࢱࢦࢽࢫ
ࢺస⏝ࡢ࣓࢝ࢽࢬ࣒㸪ᐇ㦂ື≀ࢆ⏝࠸ࡓ in vivo ࡛ࡢస⏝ࡢ᫂♧࡞࡝ࡀ௒ᚋࡢ᳨ウㄢ㢟࡜࠸࠼ࡿ㸬ࡲࡓ㸪ᮏ◊✲ࡣ㸪ࢫࢧࣅࣀࣜ⏤᮶ࡢᡂศࡀᢠ⅖⑕స⏝ࢆ᭷ࡍࡿࡇ࡜ࢆ♧ࡍ᭷ព⩏࡞⤖ᯝ࡛࠶ࡿ㸬ࢫࢧࣅࣀࣜࡢࡇࢀࡲ࡛ࡢ㣗ရ࡜ࡋ࡚ࡢ㛗ࡃ㸪㇏ᐩ࡞㣗⤒㦂ࢆ⪃៖ࡍࢀࡤ㸪ᮏ◊✲ࡢ⤖ᯝࡸࡇࢀࡲ࡛◊✲ᡂᯝ࡟ᇶ࡙ࡁ㸪Ᏻ඲࡞ᢠ⅖⑕㣗ရࢆᥦ౪࡛ࡁࡿྍ⬟ᛶࡀ㧗࠸࡜⪃࠼ࡽࢀࡿ㸬ࡋ࠿ࡋ࡞ࡀࡽ㸪ᐇ㝿࡟㣗ࡋࡓ࡜
ࡁ࡟ᢠ⅖⑕స⏝ࡀⓎ⌧ࡍࡿ࠿࡝࠺࠿ࡣ᫂☜࡟ࡉࢀ࡚࠾ࡽࡎ㸪ࡑࡢ᭷ຠᛶ࡟ࡘ࠸࡚௒ᚋ㸪ៅ㔜࡟᳨ウࡋ࡚
࠸ࡃᚲせࡀ࠶ࡿ㸬㸳㸬࠾ࢃࡾ࡟
ᮏ◊✲࡛ࡣ㸪ᶵ⬟ᛶ⣲ᮦ࡜ࡋ࡚ࡢࢫࢧࣅࣀࣜࡢ⏝㏵㛤Ⓨࢆᚿྥࡋ࡚㸪ࢫࢧࣅࣀࣜ⏤᮶⬡㉁୰ࡢᢠ⅖⑕స⏝ࢆ᭷ࡍࡿࣜࣥ⬡㉁ࡢ≉ᐃ໬ࢆヨࡳࡓ㸬ࡑࡢ⤖ᯝ㸪1) άᛶ໬࣐ࢡࣟࣇ࢓࣮ࢪ࠿ࡽࡢ NO/ࢧ࢖ࢺ࢝࢖ࣥ
⏘⏕ࢆᢚไࡍࡿ᪂つࣜࣥ⬡㉁ 1-eicosapentaenoyl2-(3-hydroxypalmitoyl)-phosphatidylglycerol(EpPOH-PG)ࢆ≉ᐃࡋࡓ㸬2) ࣜࣥ⬡㉁ࢡࣛࢫࡢ୰࡛ࡣ㸪phosphatidylglycerol ࡟ᙉ࠸ᢠ⅖⑕స⏝ࢆㄆࡵ㸪ࡲࡓ㸪୙㣬࿴⬡⫫㓟ࡀ⤖ྜࡋࡓࣜࣥ⬡㉁ࡢ᪉ࡀ㸪ࡼࡾస⏝ࡀᙉ࠸ഴྥࡀㄆࡵࡽࢀࡓ㸬 ࡞࠾㸪ᮏ◊✲࡟౑⏝ࡋࡓ࣐࢖ࢡࣟࣉ࣮ࣞࢺ࣮ࣜࢲ
࣮㸪㧗㏿ᾮయࢡ࣐ࣟࢺࢢࣛࣇ㸪࢚ࣞࢡࢺࣟࢫࣉ࣮ࣞ
࢖࢜ࣥ໬-࢖࢜ࣥࢺࣛࢵࣉᆺ㉁㔞ศᯒ⿦⨨ཬࡧ࢞ࢫࢡ࣐ࣟࢺࢢࣛࣇ࢕࣮-㉁㔞ศᯒ⿦⨨ࡣ㟁※❧ᆅᆅᇦᑐ⟇஺௜㔠ࡢ⿵ຓ࡟ࡼࡿࡶࡢ࡛࠶ࡿ㸬䋭 㪉㪎 䋭

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