Title ウシ発情周期中に発育する第１卵胞波主席卵胞の生理学的 性質 ― 発育動態、機能および受胎性に及ぼす影響 ―( 本 文(Fulltext) ) Author(s) 三浦, 亮太朗 Report No.(Doctoral Degree) 博士(獣医学) 甲第440号 Issue Date 2015-03-13 Type 博士論文 Version ETD URL http://repository.lib.gifu-u.ac.jp/handle/123456789/51006 ※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。 ࢘ࢩⓎ࿘ᮇ୰Ⓨ⫱ࡍࡿ ➨ 1 ༸⬊Ἴᖍ༸⬊ࡢ⏕⌮Ꮫⓗᛶ㉁ 㸫Ⓨ⫱ືែࠊᶵ⬟࠾ࡼࡧཷ⫾ᛶཬࡰࡍᙳ㡪㸫 2014 ᖺ ᒱ㜧ᏛᏛ㝔㐃ྜ⋇་Ꮫ◊✲⛉ 㸦ᖏᗈ␆⏘Ꮫ㸧 ୕ᾆ ுኴᮁ ┠ḟ ➨㸯❶ ⥴ゝ 1 ➨㸰❶ ➨ 1 ༸⬊Ἴᖍ༸⬊㸦W1DF㸧ࡢⓎ⫱ືែ㸪༸๓ᚋࡢᶵ⬟࠾ࡼࡧ༸ᚋ ᙧᡂࡉࢀࡿ㯤యࡢᛶ㉁ 9 2㸫1. ᗎㄽ 10 2㸫2. ヨ㦂 1㸸༸⬊ἼⓎ⌧ࡽ 7 ᪥┠ࡲ࡛ࡢ W1DF W2DF ࡢⓎ⫱ືែ࠾ࡼࡧ 10 ༸⬊ቨ⾑ὶࡢẚ㍑ 2㸫3. ヨ㦂 2㸸Ⓨ࠾ࡼࡧ༸ㄏ㉳࠾ࡅࡿ W1DF W2DF ࡢⓎ⫱ືែ㸪 ༸⬊ቨ⾑ὶ࠾ࡼࡧ⾑୰ E2 ⃰ᗘࡢẚ㍑ 14 2㸫4. ヨ㦂 3㸸W1DF W2DF ࠾ࡅࡿ༸๓ࡢ༸⬊ෆࢫࢸࣟࢻ࣍ࣝࣔࣥ⃰ᗘ ࠾ࡼࡧ㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿᅉᏊࡢẚ㍑ 16 2㸫5. ヨ㦂 4㸸W1DF W2DF ࡢ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᙧែ㸪㯤య⾑ὶ 21 ࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ࡢẚ㍑ 2㸫6. ⤖ᯝ 23 2㸫7. ⪃ᐹ 26 2㸫8. ᑠᣓ 32 ᅗ࣭⾲ 33 i ➨㸱❶ ➨ 1 ༸⬊Ἴᖍ༸⬊㸦W1DF㸧ࡢⓎ⫱ࢆไᚚࡍࡿせᅉ 3㸫1. ᗎㄽ 57 58 3㸫2. ヨ㦂 1㸸༸⬊ἼⓎ⌧ࡽ 7 ᪥┠ࡢ W1DF ࡢ┤ᚄ⾑୰ P4 ⃰ᗘࡢ┦㛵㛵ಀ ࠾ࡼࡧ W1DF 㯤యࡢඹᏑࡲࡓࡣ㠀ඹᏑ㛵ಀࡀ W1DF ࡢ┤ᚄཬࡰࡍ 58 ᙳ㡪 3㸫3. ヨ㦂 2㸸W1DF 㯤యࡢඹᏑࡲࡓࡣ㠀ඹᏑ㛵ಀࡀ༸⬊ἼⓎ⌧᪥ࡽ 5 ᪥┠ ࡲ࡛ࡢ W1DF 㯤యࡢⓎ⫱ືែ㸪⾑୰ P4 ⃰ᗘ㸪࠾ࡼࡧ༸⬊ᾮࢫࢸࣟࢻ࣍ ࣝࣔࣥ⃰ᗘཬࡰࡍᙳ㡪 60 3㸫4. ⤖ᯝ 63 3㸫5. ⪃ᐹ 64 3㸫6. ᑠᣓ 66 68 ᅗ࣭⾲ ➨㸲❶ 㯤యྠ୍༸ᕢෆᏑᅾࡍࡿ➨ 1 ༸⬊Ἴᖍ༸⬊㸦W1DF㸧ࡢᛶ㉁࠾ࡼࡧ ཷ⫾ᛶཬࡰࡍᙳ㡪 4㸫1. ᗎㄽ 77 78 4㸫2. ヨ㦂 1㸸Ἢங∵࠾ࡅࡿ W1DF 㯤యࡢඹᏑ࠾ࡼࡧ㠀ඹᏑ㛵ಀࡀ W1DF 㯤యࡢⓎ⫱ືែ࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ཬࡰࡍᙳ㡪 ii 78 4㸫3. ヨ㦂 2㸸W1DF 㯤యࡢඹᏑ࠾ࡼࡧ㠀ඹᏑ㛵ಀࡀேᕤᤵ⢭ᚋࡢཷ⫾⋡ ཬࡰࡍᙳ㡪 80 4㸫4. ⤖ᯝ 82 4㸫5. ⪃ᐹ 84 4㸫6. ᑠᣓ 87 88 ᅗ࣭⾲ ➨㸳❶ ⥲ྜ⪃ᐹ 92 ㅰ㎡ 100 ཧ⪃ᩥ⊩ 101 iii ␎ㄒ P4㸸Progesterone㸦ࣉࣟࢪ࢙ࢫࢸࣟࣥ㸧 E2㸸Estradiol-17E㸦࢚ࢫࢺࣛࢪ࣮࢜ࣝ㸧 A4㸸Androstendion㸦ࣥࢻࣟࢫࢸࣥࢪ࢜ࣥ㸧 GnRH㸸Gonadotropin releasing hormone㸦ᛶ⭢่⃭࣍ࣝࣔࣥᨺฟ࣍ࣝࣔࣥ㸧 FSH㸸Follicle stimulating hormone㸦༸⬊่⃭࣍ࣝࣔࣥ㸧 LH㸸Luteinizing hormone㸦㯤యᙧᡂ࣍ࣝࣔࣥ㸧 W1DF㸸The first follicular wave dominant follicle㸦➨ 1 ༸⬊Ἴᖍ༸⬊㸧 W2DF㸸The second follicular wave dominant follicle㸦➨ 2 ༸⬊Ἴᖍ༸⬊㸧 PG㸸Prostaglandin㸦ࣉࣟࢫࢱࢢࣛࣥࢪࣥ㸧 BFA㸸Blood flow area㸦༸⬊ቨ⾑ὶ㠃✚㸧 BF%㸸The percentage of follicle circumference with blood flow signals㸦⾑ὶศᕸྜ㸧 EIA㸸Enzyme immuno assay㸦㓝⣲ච ᐃἲ㸧 BSA㸸Bovine serum albumin㸦∵⾑Ύࣝࣈ࣑ࣥ㸧 cDNA㸸complementary DNA㸦┦⿵ⓗ DNA㸧 PCR㸸Polymerase chain reaction LHr㸸LH receptor㸦LH ཷᐜయ㸧 CYP19A1㸸Aromatase㸦࣐ࣟࢱ࣮ࢮ㸧 COX-2㸸Cyclooxygenase-2 NO㸸Nitric oxide㸦୍㓟❅⣲㸧 iNOS㸸Inducible nitric oxide synthase㸦ㄏᑟᆺ୍㓟❅⣲ྜᡂ㓝⣲㸧 eNOS㸸Endotherial nitric oxide synthase㸦⾑⟶ෆ⓶ᆺ୍㓟❅⣲ྜᡂ㓝⣲㸧 VEGF㸸Vascular endotherial growth factor㸦⾑⟶ෆ⓶ቑṪᅉᏊ㸧 FGF㸸Fibroblast growth factor㸦⥺⥔ⱆ⣽⬊ቑṪᅉᏊ㸧 iv StAR㸸Steroidogenic acute regulatory protein P450-scc㸸Cytochrome P450 side chain cleavage enzyme㸦ࢳࢺࢡ࣮࣒ࣟ P450 ഃ㙐ษ᩿㓝⣲㸧 3E-HSD㸸3E-hydroxysteroid dehydrogenase㸦3E-Ỉ㓟ᇶ⬺Ỉ⣲㓝⣲㸧 GAPDH㸸Glyceraldehyde-3-phosphate dehydrogenase㸦ࢢࣜࢭࣝࣝࢹࣄࢻ 3 ࣜࣥ㓟⬺Ỉ⣲㓝⣲㸧 W1CL㸸Corpus luteum formed after ovulation of W1DF㸦W1DF ༸ᚋ㯤య㸧 W2CL㸸Corpus luteum formed after ovulation of W2DF㸦W2DF ༸ᚋ㯤య㸧 TMR㸸Total mixed ration BCS㸸Body condition score㸦࣎ࢹ࣭ࢥࣥࢹࢩ࣭ࣙࣥࢫࢥ㸧 v ➨㸯❶ ⥴ゝ 1 ࢘ࢩࡢᐙ␆ࡣ⣙ 9,000 ᖺ๓ࡽ㛤ጞࡋࡓ⪃࠼ࡽࢀ࡚࠾ࡾ㸪ᆅ⌫ୖࡢ࠸ࡓࡿࡇࢁ ⏕ᜥࡋ࡚࠸ࡓཎ∵㸦࣮࢜ࣟࢵࢡࢫ㸪Bos primigenius㸧ࢆᇶࡋ࡚㸪すࢪᆅᇦࢆ୰ᚰ ᐙ␆ࡀ㐍⾜ࡋࡓࡉࢀࡿࠋึᮇ࠾࠸࡚㸪࢘ࢩࡣ⫗ࡢ⏝ࡸ᐀ᩍⓗᘧ⏝࠸ࡿࡇ ࢆ┠ⓗᐙ␆ࡉࢀ㸪ࡑࡢᚋࡣ㎰⪔࠾࠸࡚ࡶᚲせྍḞ࡞ᐙ␆࡞ࡗ࡚࠸ࡗࡓࠋࡋ ࡋ࡞ࡀࡽ㸪ྂ௦㑇㊧ࡽⓎぢࡉࢀࡿ㔝⏕ື≀ࡢ㦵ࡀ࢘ࢩࡢᐙ␆ࢆቃࡋ࡚ῶᑡࡍࡿ ࡇࡽ㸪ᐙ␆࢘ࢩࡢせ࡞┠ⓗࡀ㣗ᩱࢆ☜ಖࡍࡿࡓࡵ࡛࠶ࡗࡓ⪃࠼ࡽࢀ࡚࠸ࡿࠋFAO ࡢ 2007 ᖺࡢ⤫ィࡼࢀࡤ㸪 ୡ⏺࡛㣫⫱ࡉࢀ࡚࠸ࡿ࢘ࢩࡢ㢌ᩘࡣ 13 ൨ 6,000 㢌࡛࠶ࡾ㸪 ࡑࡢศᕸࡣࢪᆅᇦ 31㸣㸪୰ኸ࠾ࡼࡧ༡࣓ࣜ࢝ᆅᇦ 29㸣㸪ࣇࣜ࢝ 19㸣㸪 ࣮ࣚࣟࢵࣃ 10㸣㸪࣓ࣜ࢝ 8㸣㸪࢜ࢭࢽ 3㸣࡞ࡗ࡚࠸ࡿࠋᅜ༢࡛ࡣࣈ ࣛࢪࣝ 2 ൨㢌㸪ࣥࢻ 1 ൨ 8,000 㢌㸪࣓ࣜ࢝ྜ⾗ᅜ 9,700 㢌㸪୰ᅜ 8,200 㢌㸪ࣝࢮࣥࢳࣥࡀ 5,100 㢌⥆ࡁ㸪᪥ᮏࡣ 440 㢌ࡀᏑᅾࡍࡿࠋࡇࡢࡼ࠺ୡ ⏺୰࡛ከࡃ㣫㣴ࡉࢀ࡚࠸ࡿ࢘ࢩࡽ㸪⫗㸪⏕ஙࡉࡽຍᕤங〇ရ࡞ࡀ⏕⏘ࡉࢀ㸪࢘ࢩ ࡣ⌧௦࠾࠸࡚᭱ࡶ㔜せ࡞ᐙ␆ࡢ୍ࡘ࡛࠶ࡿࡉࢀ࡚࠸ࡿࠋ ᪥ᮏ࠾ࡅࡿᐙ␆ࡋ࡚ࡢ࢘ࢩࡢከࡃࡣ㸪⫗⏕⏘ࢆ⾜࠺⫗⏝∵㸪࠶ࡿ࠸ࡣ⏕ஙࡸங〇 ရࢆ〇㐀ࡍࡿࡓࡵࡢஙࢆ⏕⏘ࡍࡿங⏝∵ࡋ࡚㣫㣴ࡉࢀ࡚࠸ࡿࠋ⫗⏝∵ࡢ⏕⏘ࡢࡓࡵ㸪 ࡲࡓங⏝∵ࡀἪஙࡋங⏕⏘ࢆ࠾ࡇ࡞࠺ࡓࡵࡣ㸪㞤∵ࡀཷ⫾ࡋศፔࡍࡿᚲせࡀ࠶ࡾ㸪␆ ⏘ᴗࡀ⤒῭ⓗᡂ❧ࡍࡿࡓࡵࡣ㸪㞤∵ࡀຠ⋡ⓗ㡰ㄪ࡞ࢧࢡ࡛ࣝཷ⫾ࡋศፔࡋ࡚࠸ ࡃᚲせࡀ࠶ࡿࠋ ㏆ᖺ㸪᪥ᮏࢆྵࡴඛ㐍ㅖᅜ࡛ࡣ␆⏘㎰ᐙ㸪≉㓗㎰ᐙ 1 ᡞᙜࡓࡾࡢ㣫㣴㢌ᩘࡀቑຍࡋ ࡚࠸ࡿࠋ᪥ᮏࢆ࠶ࡆࡿ㸪1 ᡞᙜࡓࡾࡢங∵㣫㣴㢌ᩘࡣ 1960 ᖺࡣ 2.0 㢌࡛࠶ࡗࡓ ࡢᑐࡋ࡚㸪1980 ᖺࡣ 18.1 㢌㸪1990 ᖺࡣ 32.5 㢌㸪2000 ᖺࡣ 52.5 㢌ࡑࡋ࡚ 2012 ᖺࡣ 72.1 㢌࡞ࡾ㸪ࡇࡢ 50 ᖺ࡛ᛴ⃭࡞㣫㣴つᶍᣑࡀࡳࡽࢀࡿ㸦␆⏘⤫ィㄪᰝ㸪㎰ ᯘỈ⏘┬㸧 ࠋࡲࡓ㸪࢘ࢩࡢ㑇ఏᨵⰋࡸ㣫㣴⟶⌮ᢏ⾡ࡢྥୖࡼࡾ㸪ங⏝∵ࡢಶయங㔞ࡣ ୡ⏺ⓗቑຍഴྥ࠶ࡿࠋ࣓ࣜ࢝ྜ⾗ᅜ࡛ࡣ 1951 ᖺࡽ 1996 ᖺࡅ࡚ 5,000 kg 2 ࡽ 9,000 kg ௨ୖᖺ㛫ᖹᆒಶయங㔞ࡀቑຍࡋ࡚࠸ࡿ㸦18㸧 ࠋ᪥ᮏ࠾ࡅࡿங⏝∵ࡢᖺ㛫 ᖹᆒಶయங㔞ࡣ㸪1975 ᖺࡣ 5,826 kg ࡛࠶ࡗࡓࡢᑐࡋ࡚㸪1980 ᖺࡣ 6,339 kg㸪1990 ᖺࡣ 7,798 kg㸪2000 ᖺࡣ 8,794 kg ࡑࡋ࡚ 2010 ᖺࡣ 9,286 kg ࡞ࡾⴭࡋࡃቑຍࡋ࡚ ࠸ࡿ㸦ங⏝∵⩌⬟ຊ᳨ᐃᡂ⦼ࡢࡲࡵ㸫ᖹᡂ 24 ᖺᗘ㸫㸪ᐙ␆ᨵⰋᴗᅋ㸧 ࠋࡲࡓ㸪⫗⏝ ∵࠾࠸࡚ࡶ 1 ᡞᙜࡓࡾࡢ㣫㣴㢌ᩘࡣ 1960 ᖺ࡛ࡣ 1.2 㢌࡛࠶ࡗࡓࡢᑐࡋ࡚㸪1980 ᖺ ࡣ 5.9 㢌㸪1990 ᖺࡣ 11.6 㢌㸪2000 ᖺࡣ 24.2 㢌ࡑࡋ࡚ 2012 ᖺࡣ 41.8 㢌࡞ࡾ㸪ங ⏝∵ྠᵝ㣫㣴つᶍᣑࡀぢཷࡅࡽࢀࡿ㸦␆⏘⤫ィㄪᰝ㸪㎰ᯘỈ⏘┬㸧 ࠋ ⌧௦࠾࠸࡚㸪ከࡃࡢ㞤∵ࡣேᕤᤵ⢭ࡼࡾཷ⫾㸪ศፔࡋ࡚࠸ࡿࡇࡀ▱ࡽࢀ࡚࠾ࡾ㸪 ேᕤᤵ⢭ࡢᬑཬ⋡㛵ࡋ࡚ࡣ㸪࣓ࣜ࢝ྜ⾗ᅜ࡛ࡣங⏝∵ࡢ 72㸣㸦92㸧 㸪᪥ᮏ࡛ࡣங⏝ ∵㸪⫗⏝∵ࡶ 95㸣௨ୖ㸦44㸧࡞ࡗ࡚࠸ࡿࠋࡇࡢࡓࡵ㸪ຠ⋡ⓗࡘ㐺ษ࡞㞤∵ࡢⓎ Ⓨぢࡀ⦾Ṫᡂ⦼ྥୖࡢ㔜せ࡞せᅉ࡞ࡗ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽ㸪ᛴ⃭࡞㣫㣴つᶍࡢᣑ క࠺㸪ⓎⓎぢ⋡ࡢపୗࡀ✵⫾ᮇ㛫ࡢᘏ㛗ࡘ࡞ࡀࡾ㸪⦾Ṫᡂ⦼ࡢపୗࢆࡶࡓࡽࡋ ࡚࠸ࡿࠋࡲࡓ㸪ங⏝∵㛵ࡋ࡚ࡣ㧗Ἢஙక࠸㸪⫢⮚࡛ࡢࢫࢸࣟࢻ௦ㅰஹ㐍㸦100㸧 ࡀᣦࡉࢀ࡚࠾ࡾ㸪ࡑࢀక࠺Ⓨᚤᙅ࠾ࡼࡧⓎⓎ⌧㛫ࡢ▷⦰㸦55㸧㉳ᅉࡍࡿ ⓎⓎぢ⋡࡞ࡢపୗࡸ㸪㐺ᮇᤵ⢭ࡀᐇ࡛ࡁ࡞࠸ࡇࡼࡿཷ⫾ᛶࡢపୗࡀࡁ࡞ၥ 㢟࡞ࡗ࡚࠸ࡿࠋ ࡑࡢࡓࡵ㸪࣓ࣜ࢝ྜ⾗ᅜࢆ୰ᚰࡋࡓඛ㐍ᅜ࡛ࡣ㸪⦾Ṫᡂ⦼ྥୖࢆ┠ⓗྛ✀࣍ࣝ ࣔࣥ〇ࢆ⏝࠸࡚㸪࢜ࣈࢩࣥࢡἲ㸦72㸧࡞ⓎⓎぢࢆᚲせࡋ࡞࠸ᐃேᕤᤵ⢭ࣉࣟ ࢢ࣒ࣛࡢ㛤ⓎᨵⰋࢆ⾜ࡗ࡚ࡁࡓ㸦32㸪50㸪60㸪77㸪90㸧 ࠋࡋࡋ࡞ࡀࡽ㸪ࡇࢀࡽࡢࣉࣟ ࢢ࣒ࣛࢆ⏝࠸ࡿࡇ࡛㸪ᤵ⢭⋡ࢆྥୖࡉࡏࡿࡇࡣᡂຌࡋࡓࡀ㸪ཷ⫾⋡ࡢྥୖ㛵ࡋ ࡚ࡣ⌧ᅾ⮳ࡿࡲ࡛༑ศ㐩ᡂ࡛ࡁ࡚࠸࡞࠸ࠋࡇࡢࡇࡣ㸪࢘ࢩࡢ⦾Ṫ⏕⌮ᑐࡍࡿ ⌮ゎࡸ㸪ዷፎᡂ❧㛵ࢃࡿせᅉࡢゎ᫂ࡉࡽࡣཷ⫾⋡ࡢྥୖࢆࡶࡓࡽࡍෆศἪ≧ែࢆస ฟࡍࡿᤵ⢭ࣉࣟࢢ࣒ࣛࡢ㛤Ⓨࡀ☜❧ࡋ࡚࠸࡞࠸ࡇࢆ♧၀ࡋ࡚࠸ࡿࠋࡑࡢࡓࡵ㸪࢘ࢩࡢ ⦾Ṫ⏕⌮ᑐࡋ࡚ࡲ࡛ὀ┠ࡉࢀ࡚࠸࡞ࡗࡓⅬࡘ࠸࡚ࡢᇶ♏ⓗ࡞ㄪᰝࡸ◊✲ࢆ⾜ࡗ 3 ࡚࠸ࡃᚲせࡀ࠶ࡿࠋ 㞤ࡀ㓄ࡢࡓࡵ㞝ࢆチᐜࡍࡿ≧ែࡀⓎᐃ⩏ࡉࢀ㸪ࡇࡢⓎࡣ࿘ᮇⓗ⧞ࡾ㏉ࡉ ࢀ࡚࠾ࡾ㸪ࡇࡢ࿘ᮇࢆⓎ࿘ᮇ࠸࠺ࠋ࢘ࢩࡣ࿘ᖺ⦾ṪࡢከⓎື≀࡛࠶ࡾ㸪6㹼12 ࣧ ᭶㱋࡛ᛶᡂ⇍㐩ࡋࡓᚋ㸦30㸧 㸪ዷፎࡋ࡞࠸㝈ࡾ 1 ᖺࢆ㏻ࡋ࡚࿘ᮇⓗⓎࢆ⧞ࡾ㏉ࡍࡼ ࠺࡞ࡿࠋⓎ࿘ᮇ୰ࡣ༸ᕢෆᵓ㐀≀࡛࠶ࡿ㯤య༸⬊ࡢⓎ⫱㛢㙐࣭㏥⾜ࡢ࿘ᮇⓗ ኚࡀࡳࡽࢀࡿࠋ࢘ࢩࡢⓎ࿘ᮇࡢᖹᆒ᪥ᩘࡣ 21 ᪥࡛࠶ࡿ㸦30㸧ࡀ㸪Ⓨ࿘ᮇࡢ᪥ᩘ ࡣ 18㹼24 ᪥ᖜࡢ࠶ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ㸦30㸧 ࠋⓎ࿘ᮇࡣࡁࡃ㯤యᮇ㸦14㹼18 ᪥ 㛫㸧༸⬊ᮇ㸦4㹼6 ᪥㛫㸧ࡢ 2 ࡘศ㢮ࡉࢀࡿ㸦30㸧 ࠋ㯤యᮇࡣⓎᚋ༸ᕢෆ࡛ᡂ⇍ ༸⬊ࡀ༸ࡋࡓ┤ᚋࡽጞࡲࡾ㸪㯤యࡀᡂ⇍ࡋࡓᚋ㸪㯤య㏥⾜ࡀ㛤ጞࡍࡿࡲ࡛ࡢᮇ㛫࡛ ࠶ࡾ㸪㯤య㏥⾜ࡀ㛤ጞࡋ࡚ࡽ༸⮳ࡿࡲ࡛ࡢᮇ㛫ࢆ༸⬊ᮇ࠸࠺㸦30㸧 ࠋ༸⬊ᮇࡢᡂ ⇍༸⬊ࡽ࢚ࢫࢺࣛࢪ࣮࢜ࣝ㸦E2㸧ࡀ⏘⏕࠾ࡼࡧศἪࡉࢀࡿ㸦30㸧ࡇ࡛⾑୰ E2 ⃰ᗘࡀ ୖ᪼ࡋ㸪ṇࡢࣇ࣮ࢻࣂࢵࢡస⏝ࡼࡾどᗋୗ㒊࡛ᛶ⭢่⃭࣍ࣝࣔࣥᨺฟ࣍ࣝࣔࣥ 㸦GnRH㸧ࡢࢧ࣮ࢪ≧ศἪࡀㄏ㉳ࡉࢀ㸪ࡑࢀࡀ⬻ୗᆶయ๓ⴥస⏝ࡍࡿࡇ࡛㸪㯤యᙧ ᡂ࣍ࣝࣔࣥ㸦LH㸧ࡢࢧ࣮ࢪ≧ࡢᨺฟࡀ㉳ࡇࡾ㸦80㸧 㸪ᡂ⇍༸⬊ࡢ༸ࢆㄏᑟࡍࡿ㸦94㸧 ࠋ ᡂ⇍༸⬊ࢆᵓᡂࡋ࡚࠸ࡿ㢛⢏ᒙ⣽⬊ෆ༸⬊⭷⣽⬊ࡀ LH ࢧ࣮ࢪࡼࡾ่⃭ࡉࢀ㸪ࡑࢀ ࡒࢀᆺ㯤య⣽⬊ᑠᆺ㯤య⣽⬊ኚࡋ㸪ࡑࢀࡽࡀቑṪࡍࡿࡇ࡛ᛴ㏿㯤యࡀᙧᡂ ࡉࢀ࡚࠸ࡃ㸦8㸧 ࠋ㯤యࡣ㸪Ⓨ࿘ᮇࡢ 7 ᪥┠ࡈࢁࡲ࡛Ⓨ⫱㸪ᡂ㛗ࡋ࡚࠸ࡁ㸪ࡑࡢᚋࡣ㯤 య㏥⾜㛤ጞࡲ࡛ᶵ⬟ⓗ㯤యࡋ࡚Ꮡᅾࡍࡿࠋዷፎࡀᡂ❧ࡋ࡞࠸ሙྜ㸪Ꮚᐑࡽࣉࣟࢫࢱ ࢢࣛࣥࢪࣥ㸦PG㸧F2Dࡀᨺฟࡉࢀࡿ㸦54㸧ࡇ࡛㯤య㏥⾜ࡀ㛤ጞࡉࢀ㸪㯤యࡣᛴ⃭⦰ ᑠࡍࡿ㸦11㸪51㸪67㸧 ࠋ㯤యࡽࡣ㸪ࣉࣟࢪ࢙ࢫࢸࣟࣥ㸦P4㸧ࡀ⏘⏕㸪ศἪࡉࢀ࡚࠾ࡾ㸦68㸧 㸪 㯤యࡢⓎ⫱క࠸⾑୰ P4 ⃰ᗘࡣୖ᪼ࡋ࡚࠸ࡁ㸪㯤యᙧᡂࡀࡍࡿ㯤య㏥⾜㛤ጞࡲ࡛ ⾑୰ P4 ⃰ᗘࡣ㧗ࡃ⥔ᣢࡉࢀ㸪 㯤య㏥⾜㛤ጞక࠸ᛴ⃭⾑୰ P4 ⃰ᗘࡣపୗࡋ࡚࠸ࡃ 㸦57㸧 ࠋ ࡇࡢࡼ࠺࡞㯤యࡢⓎ⫱㸪⥔ᣢ࠾ࡼࡧ㏥⾜㸪༸⬊ࡢᡂ⇍༸ࡀ⧞ࡾ㏉ࡉࢀ㸪Ⓨ࿘ᮇ ࡀႠࡲࢀࡿࠋࡇࡢⓎ࿘ᮇᮇ㛫୰ࡣ༸ᕢෆ࡛ࡣ 2㹼3 ᅇࡢ༸⬊⩌ࡢⓎ⫱ࡀほᐹࡉࢀ㸪ࡑ 4 ࢀࡒࢀࡢ༸⬊⩌ࡽ༸⮳ࡽ࡞࠸༸⬊ࡀ 1㹼2 ࡘⓎ⫱ࡍࡿࡇࡀ▱ࡽࢀ࡚࠾ࡾ㸪ࡇࡢ༸ ⬊Ⓨ⫱ࡢ୍⩌ࢆ༸⬊Ἴ࠸࠺㸦41㸧 ࠋ ࢘ࢩ࠾࠸࡚Ⓨ࿘ᮇ୰Ⓨ⌧ࡍࡿ᭱ึࡢ༸⬊Ἴࡣ㸪Ⓨ⤊ᚋࡢ༸ᕢෆ࡛ 3㹼4mm ⛬ࡢᑠ༸⬊ࡢ୍⩌㸦8㹼41 ಶ㸧ࡢⓎ⫱ࡀ㛤ጞࡉࢀ㸪ࡇࢀࢆ➨ 1 ༸⬊Ἴ࠸࠺㸦41㸧 ࠋࡑࡢ ୍⩌ࡢ༸⬊ࡢ୰࡛᭱ࡢ༸⬊ࡀ 1 ࡘࡔࡅ㑅ᢤࡉࢀ㸪ࡑࢀ௨እࡢᑠ༸⬊ࡣ㛢㙐ࡍࡿࠋࡇࡢ 㑅ᢤࡉࢀࡓ༸⬊ࢆᖍ༸⬊࠸࠸㸪 ➨ 1 ༸⬊Ἴࡢᖍ༸⬊ࢆ➨ 1 ༸⬊Ἴᖍ༸⬊ 㸦W1DF㸧 ࠸࠺ࠋW1DF ࡣⓎ࿘ᮇࡢ 8㹼11 ᪥㡭ࡲ࡛Ⓨ⫱ࡍࡿࡀ㸪㯤యࡽ P4 ࡀ⏘⏕ศἪࡉࢀ࡚ ࠸ࡿᮇᙜࡓࡿࡓࡵ㸪ᡂ⇍ࡍࡿࡇࡀ࡛ࡁࡎ㸪ࡇࡢ W1DF ࡣ༸⮳ࡽࡎ㛢㙐ࡋ࡚ࡋ ࡲ࠺㸦41㸧 ࠋࡑࢀక࠸᪂ࡓ࡞༸⬊ἼࡢⓎ⌧ࡀ㛤ጞࡉࢀ㸪ࡇࢀࢆ➨ 2 ༸⬊Ἴ࠸࠺ࠋ࢘ࢩ ࡣ༸⬊Ἴࢆ 2 ࡘᣢࡘࡶࡢ㸪3 ࡘᣢࡘࡶࡢࡀᏑᅾࡋ㸪ࡑࢀࡒࢀࡢⓎ⏕⋡ࡣヨ㦂⏝࠸ ࡿ࢘ࢩࡢရ✀࠾ࡼࡧᖺ㱋ࡼࡾ␗࡞ࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸦7㸪16㸪17㸪19㸪28㸪39㸪 69㸪74㸪88㸪108㸧 ࠋ༸⬊Ἴࢆ 3 ࡘᣢࡘ࢘ࢩࡢẚ⋡ࡣపᰤ㣴≧ែ㸦66㸪76㸧ࡸࣄ࣮ࢺࢫࢺ ࣞࢫ᮲௳ୗ㸦10㸧࡛ቑຍࡍࡿ࠸࠺ሗ࿌ࡀ࠶ࡿࠋW1DF ࡢ⏕Ꮡᮇ㛫ࡀ༸⬊Ἴࡢᩘᙳ㡪 ࢆ࠼ࡿྍ⬟ᛶࢆ᥎ ࡋ࡚࠸ࡿሗ࿌㸦48㸧ࡶ࠶ࡿࡀ㸪ヲ⣽ࡣ᫂ࡽࡉࢀ࡚࠸࡞࠸ࠋ༸ ⬊Ἴࢆ 2 ࡘᣢࡘ࢘ࢩ࡛ࡣ㸪➨ 2 ༸⬊ἼࡢⓎ⌧᪥ࡣⓎ࿘ᮇࡢ 9㹼11 ᪥㡭࡛࠶ࡾ㸪༸⬊Ἴ ࢆ 3 ࡘᣢࡘ࢘ࢩ࡛ࡣ 8㹼9 ᪥㡭࡛࠶ࡿࡉࢀ࡚࠸ࡿ㸦4㸧 ࠋ༸⬊Ἴࢆ 3 ࡘᣢࡘ࢘ࢩ࡛ࡣ㸪➨ 3 ༸⬊ἼࡢⓎ⌧᪥ࡀ 15㹼16 ᪥㡭࡛࠶ࡿ㸦4㸧 ࠋࡲࡓ㸪༸⬊Ἴࢆ 2 ࡘᣢࡘ࢘ࢩ 3 ࡘᣢࡘ࢘ ࢩ࡛㯤య㏥⾜㛤ጞᮇⓎ࿘ᮇ᪥ᩘࢆẚ㍑ࡋࡓࡇࢁ㸪㯤య㏥⾜㛤ጞᮇࡣ༸⬊Ἴࢆ 2 ࡘᣢࡘ࢘ࢩ࡛᪩ᮇ㛤ጞࡉࢀ㸦2 ༸⬊Ἴ vs. 3 ༸⬊Ἴ㸻16 ᪥┠ vs. 19 ᪥┠㸧 㸪ࡑࡢ⤖ᯝ㸪 Ⓨ࿘ᮇ᪥ᩘࡀ▷⦰ࡍࡿ㸦2 ༸⬊Ἴ vs. 3 ༸⬊Ἴ㸻19㹼20 ᪥┠ vs. 22㹼23 ᪥┠㸧 㸦4㸧 ࠋ༸ ⬊Ἴࢆ 2 ࡘ࠶ࡿ࠸ࡣ 3 ࡘᣢࡘ࢘ࢩࡢࡕࡽࡢሙྜ࠾࠸࡚ࡶ㸪➨ 1 ༸⬊ἼࡢⓎ⌧ᮇࡣ ༸┤ᚋ࡛࠶ࡾ㸪㑅ᢤࡉࢀࡓᖍ༸⬊ࡣ༸ࡏࡎ㸪㛢㙐ࡍࡿ࠸࠺Ⅼ࡛ඹ㏻ࡋ࡚࠸ࡿࡓ ࡵ㸪➨ 1 ༸⬊Ἴࢆ⏝࠸ࡓ༸⬊Ⓨ⫱㛵ࡍࡿ◊✲ࡣከ࠸ࠋࡋࡋ㸪ࡇࡢ➨ 1 ༸⬊Ἴࡀ࢘ࢩ ࡢ⦾Ṫ⏕⌮࠾࠸࡚ᯝࡓࡍᙺࡸព⩏㛵ࡍࡿ◊✲ࡣᑡ࡞࠸ࠋ 5 㞤ࡢ⏕ṪෆศἪ⣔ࡣどᗋୗ㒊㸫ୗᆶయ㸫༸ᕢ㍈ࢆ୰ᚰᵓᡂࡉࢀ࡚࠾ࡾ㸪どᗋୗ㒊࡛ ࡣ GnRH ࡢศἪࡀ⾜ࢃࢀ㸪ୗᆶయ๓ⴥ࡛ࡣ༸⬊่⃭࣍ࣝࣔࣥ㸦FSH㸧 LH ࡀศἪࡉࢀ ࡿ㸦80㸧 ࠋどᗋୗ㒊ࡢṇ୰㝯㉳㒊ࡽᨺฟࡉࢀࡓ GnRH ࡣୗᆶయ㛛⬦ධࡾ㸦65㸧 㸪ୗᆶ య๓ⴥస⏝ࡋ㸪FSH ࠾ࡼࡧ LH ࡢྜᡂศἪࡀಁ㐍ࡉࢀࡿࠋFSH ࡣ༸⬊ࡢⓎ⫱ࡸ༸⬊ ᩘࡢㄪ⠇㛵ࡍࡿ࡛࣍ࣝࣔࣥ࠶ࡾ㸪༸⬊ἼⓎ⌧㛵ࢃࡿ㸦6㸧 ࠋLH ࡢࣃࣝࢫ≧ศἪࡣ GnRH ࡢᢿືᛶศἪ㐃ືࡋ࡚࠾ࡾ㸦107㸧 㸪LH ࡣᖍ༸⬊ࡢ㑅ᢤᖍ༸⬊ࡢⓎ⫱㸪ࡉ ࡽᡂ⇍㛵ࢃࡿ࡛࣍ࣝࣔࣥ࠶ࡿ㸦36㸧 ࠋどᗋୗ㒊ࡢάືࡣ㸪༸ᕢࡽศἪࡉࢀࡿᛶࢫࢸ ࣟࢻ࣍ࣝࣔࣥ㸦E2 ࠾ࡼࡧ P4㸧ࡼࡾࣇ࣮ࢻࣂࢵࢡㄪ⠇ࡉࢀ࡚࠸ࡿࠋ㐣ཤࡢሗ࿌࠾ ࠸࡚㸪⾑୰ P4 ⃰ᗘࡀప࠸ LH ࡢࣃࣝࢫ㢖ᗘࡀቑຍࡍࡿࡇ㸪୍᪉࡛⾑୰ P4 ⃰ᗘࡀ㧗 ࠸ LH ࡢࣃࣝࢫ㢖ᗘࡀῶᑡࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ㸦15㸪52㸧 ࠋLH ࡢࣃࣝࢫ≧ศἪ ࡣᖍ༸⬊ࡢⓎ⫱ᚲせ࡛࠶ࡿࡓࡵ㸪⾑୰ P4 ⃰ᗘࡀ㧗⃰ᗘ࡞ࡿ㯤యᮇ࡛ࡣ㸪ᖍ༸⬊ ࡢⓎ⫱ࡣᢚไࡉࢀ༸⮳ࡿࡇࡣ࡞ࡃ㛢㙐ࡋ࡚࠸ࡃࠋࡋࡋ࡞ࡀࡽ㸪㯤యࡀ㏥⾜ࡋ⾑ ୰ P4 ⃰ᗘࡀప࠸≧ἣ࡛ࡣ㸪LH ࣃࣝࢫ㢖ᗘࡀୖ᪼ࡍࡿࡇ࡛㸪༸⬊ࡢⓎ⫱ࡀಁࡉࢀ㸪ᡂ ⇍༸⬊࡛⏘⏕ࡉࢀࡓ E2 ࡀศἪࡉࢀ㸪⾑୰ E2 ⃰ᗘࡀୖ᪼ࡍࡿࠋ⾑୰ E2 ⃰ᗘࡢୖ᪼どᗋ ୗ㒊ࡀᛂࡋ GnRH ࢧ࣮ࢪࡀᘬࡁ㉳ࡇࡉࢀ㸪ࡑࢀᛂࡋ࡚ୗᆶయ๓ⴥ࡛ࡣ LH ࢧ࣮ࢪ ࡀㄏ㉳ࡉࢀ㸪ᡂ⇍༸⬊ࡣ༸ࡍࡿ㸦81㸧 ࠋ ⫾⏕ᮇ࠾ࡼࡧᶵⓎື๓ࡢ࢘ࢩࡢ༸ᕢ࠾࠸୍࡚ḟ༸ẕ⣽⬊ࡢ࿘ᅖ༸⬊ୖ⓶⣽⬊࡛ ⿕ࢃࢀࡓཎጞ༸⬊ࡀᙧᡂࡉࢀࡿࠋࡑࡢᚋ㸪ᶵⓎືక࠸㸪ཎጞ༸⬊ࡣⓎ⫱ࢆ㛤ጞࡋ㸪 ࡇࡢⓎ⫱ࢆ㛤ጞࡋࡓ༸⬊ࡣ୍ḟ༸⬊ࡼࡤࢀࡿࠋ୍ḟ༸ẕ⣽⬊ࢆྲྀࡾᅖࢇ࡛࠸ࡿ༸⬊ୖ ⓶⣽⬊ࡣయ✚ࡀቑࡋ❧᪉≧࡞ࡿ㸦63㸧 ࠋࡉࡽ❧᪉ࡋࡓୖ⓶⣽⬊ࡀቑṪ㸪㔜ᒙࡍࡿ ࡇ࡛㢛⢏ᒙ⣽⬊࡞ࡾ㸪ࡉࡽࡑࡢእഃࢆෆ༸⬊⭷⣽⬊ࡀྲྀࡾᅖࢇࡔ≧ែ࡞ࡾḟ ༸⬊࡞ࡿ㸦49㸧 ࠋࡇࡢẁ㝵ࡲ࡛ࡣ㸪༸⬊ࡣᛶ⭢่⃭࣍ࣝࣔࣥ㸦FSH ࠾ࡼࡧ LH㸧౫Ꮡ ࡍࡿࡇ࡞ࡃⓎ⫱ࡀ㐍⾜ࡍࡿࠋᆺ༸⬊ࡢᶵ⬟ࡢᾘኻ㸪ࡘࡲࡾⓎᚋࡢ༸ࡸ➨ 1 ࠶ࡿ ࠸ࡣ➨ 2 ༸⬊Ἴᖍ༸⬊㸦W2DF㸧ࡢ㛢㙐ࡀ㉳ࡇࡿ㸪ᖍ༸⬊ࡢ㢛⢏ᒙ⣽⬊࡛⏘⏕ࡉ 6 ࢀࡿࣥࣄࣅࣥࡢ⾑୰⃰ᗘࡀపୗࡍࡿࠋࣥࣄࣅࣥࡣୗᆶయ๓ⴥస⏝ࡋ FSH ࡢᨺฟࢆ ᢚไࡍࡿࡓࡵ㸪༸ᚋࡸᖍ༸⬊㛢㙐㛤ጞࡶ FSH ࡢࢧ࣮ࢪ≧ศἪࡀ㉳ࡇࡿ㸦62㸧 ࠋ ࡑࢀࢆཷࡅ࡚ḟ༸⬊ࡢ୰ࡽᩘࡋ࡚ 8㹼41 ಶ㸪┤ᚄ 3㹼4mm ⛬ࡢ༸⬊ࡢⓎ⫱ࡀ㛤ጞ ࡉࢀ㸪༸⬊ἼࡢⓎ⌧࡞ࡿ㸦17㸪34㸧 ࠋ༸⬊Ἴࡋ࡚ືဨࡉࢀࡓ༸⬊ࡣᛶ⭢่⃭࣍ࣝࣔࣥ ౫ᏑⓗⓎ⫱ࢆ⥆ࡅࡿࠋࡇࡢ୍⩌ࡢ༸⬊ࡣ 1 ᪥ᙜࡓࡾ 1.5㹼2.0 mm ࡢ㏿ᗘ࡛Ⓨ⫱ࡋ࡚࠸ ࡃ㸦37㸧 ࠋ༸⬊ἼࡢⓎ⌧ࡽ 2㹼3 ᪥⤒㐣ࡍࡿ㸪୍⩌ࡢ୰࡛᭱༸⬊ࡢ㢛⢏ᒙ⣽⬊࡚ ࠋE2 ࡣୗᆶయ๓ⴥస⏝ࡋ FSH LH ཷᐜయࡀⓎ⌧ࡋ㸪E2 ⏘⏕ࡀ⾜ࢃࢀࡿࡼ࠺࡞ࡿ㸦13㸧 ࡢᨺฟࢆᢚไࡍࡿ㸦38㸧 㸪ࡑࡢ⤖ᯝ㸪༸⬊Ἴࡢ༸⬊⩌ࡢ୰࡛ᖍ༸⬊ࡋ࡚㑅ᢤࡉࢀ࡞ ࡗࡓḟᖍ௨ୗࡢ FSH ౫Ꮡᛶࡢ༸⬊ࡣⓎ⫱ࡀ⥔ᣢ࡛ࡁࡎ㛢㙐ࡋ࡚࠸ࡃࠋࡋࡋ࡞ࡀࡽ㸪᭱ ༸⬊ࡣ㢛⢏ᒙ⣽⬊ LH ཷᐜయࢆⓎ⌧ࡍࡿࡇ࡛㸪ࡑࡢⓎ⫱ࢆ FSH ౫Ꮡᛶࡽ LH ౫ Ꮡᛶࢩࣇࢺࡍࡿࡓࡵ㸪LH ࣃࣝࢫࡢ่⃭ࢆཷࡅ࡚㸪㛢㙐ࡍࡿࡇ࡞ࡃⓎ⫱ࡀ⥔ᣢࡉࢀ ࡿࠋࡇࡢ㐣⛬ࢆ㑅ᢤ࠸࠸㸪㑅ᢤࡉࢀࡓ᭱༸⬊ࢆᖍ༸⬊࠸࠺㸦43㸧 ࠋࡇࡢ㑅ᢤ ࠾ࡅࡿ᭱༸⬊ࡢᖹᆒ┤ᚄࡣ 8.5mm ࡛࠶ࡾ㸪ḟᖍ༸⬊ࡢᖹᆒ┤ᚄࡣ 7.7mm ࡉࢀ࡚࠸ ࡿ㸦35㸧 ࠋW1DF ࡣ㸪㯤యࡀᙧᡂࡉࢀ㸪⾑୰ P4 ⃰ᗘࡀୖ᪼ࡍࡿᮇⓎ⫱ࡍࡿࡓࡵ㸪 ࢇࡀ༸⮳ࡿࡇ࡞ࡃ㛢㙐ࡋ࡚ࡋࡲ࠺ࠋ୍᪉࡛㸪W2DF ࡛ࡣ㸪Ⓨ⫱ࡋ࡚࠸ࡿᮇ㛫 㯤యࡢ㏥⾜ࡀ㉳ࡇࡾ⾑୰ P4 ⃰ᗘࡀపୗࡋ㸪LH ࣃࣝࢫ㢖ᗘࡀୖ᪼ࡍࡿࡇ࡛ᖍ༸⬊ ࡢⓎ⫱ࡀ⥔ᣢࡉࢀ㸪༸⬊ࡽከ㔞ࡢ E2 ࡀ⏘⏕࠾ࡼࡧศἪࡉࢀࡿࠋࡑࡢ⤖ᯝ㸪LH ࢧ࣮ࢪ ࡀᘬࡁ㉳ࡇࡉࢀ㸪W2DF ࡣᡂ⇍ࡋ༸⮳ࡿࠋ ࡇࡢ➨ 1 ༸⬊Ἴ࠾࠸࡚㸪ࡑࡢⓎ⫱ືែࡸ࣍ࣝࣔࣥືែ㛵ࡋ࡚ヲ⣽ㄪᰝࡉࢀ࡚ࡣ ࠸ࡿࡀ㸪W1DF ⮬㌟ࡢᛶ㉁ࡸ㸪ࡑࢀࡀ⦾Ṫ⏕⌮ࡸཷ⫾ᛶཬࡰࡍᙳ㡪㛵ࡋ࡚ࡣ༑ศ ᳨ウࡉࢀ࡚ࡣ࠸࡞࠸ࠋ᪤㏙ࡓࡼ࠺㸪➨ 1 ༸⬊Ἴࡣ㯤యᙧᡂඹⓎ⫱ࡍࡿࡓࡵ㸪 W1DF ࡣ⾑୰ P4 ⃰ᗘࡀప್ࡽᚎࠎୖ᪼ࡍࡿᮇᡂ㛗ࡍࡿࠋ୍᪉㸪➨ 2 ༸⬊Ἴࡣ㯤 యࡀ༑ศᡂ⇍ࡋ࡚ࡽⓎ⫱ࢆ㛤ጞࡍࡿࡓࡵ㸪W2DF ࡣ⾑୰ P4 ⃰ᗘࡀ㧗್࡞ᮇᡂ㛗 ࡍࡿࠋᖍ༸⬊ࡢⓎ⫱ࡣ⬻ୗᆶయ๓ⴥࡽศἪࡉࢀࡿ LH ࡢࣃࣝࢫ≧ࡢศἪࡀᚲ㡲࡛ 7 ࠶ࡿࡓࡵ㸪W1DF W2DF ࡀⓎ⫱ࡍࡿᮇ㛫࡛ࡢ⾑୰ P4 ⃰ᗘ᥎⛣ࡀ␗࡞ࡿࡇࡼࡿ㸪 LH ࣃࣝࢫ㢖ᗘࡢ㐪࠸ࡀⓎ⫱ࡍࡿᖍ༸⬊ࡢᙧែࡸᶵ⬟ᕪ␗ࢆࡶࡓࡽࡍ⪃࠼ࡽࢀࡿࠋ 㐣ཤࡢ◊✲ࢆࡳ࡚ࡶ㸪W1DF W2DF ࡢᛶ㉁ࡸᶵ⬟ࡢ㐪࠸ࢆ༑ศẚ㍑ࡋࡓ◊✲ࡣ ࢇ⾜ࢃࢀ࡚ࡇ࡞ࡗࡓࠋࡑࡇ࡛㸪W1DF ࡢ≉᭷ࡢᛶ㉁ࡸᶵ⬟ࢆ᫂ࡽࡍࡿࡓࡵ㸪 W1DF W2DF ࡢᛶ㉁ࢆẚ㍑ࡋࡓࠋ ࡲࡓ㸪W1DF ࡢⓎ⫱ࢆไᚚࡍࡿせᅉࢆ᫂ࡽࡍࡿࡓࡵ㸪➨ 1 ༸⬊ἼⓎ⫱ᮇ㛫୰ ࠾ࡅࡿᛶࢫࢸࣟࢻ࣍ࣝࣔࣥືែࡸ㸪༸ᕢෆ࡛Ⓨ⫱ࡍࡿ㯤యࡢ⨨㛵ಀ࡞ࡢせᅉ W1DF ࡢᛶ㉁ࢆẚ㍑ࡍࡿࡇࡋࡓࠋࡉࡽ㸪㐣ཤࡢሗ࿌ࡽ㯤యࡀᏑᅾࡍࡿ༸ᕢྠ ഃࡢᏊᐑゅࡢෆ⭷୰ࡢ P4 ⃰ᗘࡀ㧗࠸ࡇ㸦20㸧 㸪༸๓༸⬊ࡀᏑᅾࡍࡿ༸ᕢྠഃࡢ༸ ⟶୰ࡢ E2 ⃰ᗘࡀ㧗࠸ࡇ㸦99㸧ࡀ▱ࡽࢀ࡚࠾ࡾ㸪ᒁᡤ࡛ࡢ༸ᕢᏊᐑゅࡢᐦ᥋࡞㛵㐃ᛶ ࡀᏑᅾࡍࡿࡇࡀ♧၀ࡉࢀࡿࠋࡋࡋ࡞ࡀࡽ㸪W1DF ࡀྠ୍༸ᕢෆࡢ㯤యࡸᏊᐑゅᑐ ࡋ࡚ᒁᡤⓗࡢࡼ࠺࡞ᙳ㡪ࢆ࠼ࡿࡢࢆㄪᰝࡋࡓ◊✲ࡣぢᙜࡓࡽ࡞࠸ࠋ ᮏ◊✲࠾࠸࡚㸪➨ 2 ❶࡛ࡣ㸪W1DF ࡢᛶ㉁࠾ࡼࡧᶵ⬟ࢆ᫂ࡽࡍࡿࡓࡵ㸪㠀Ἢங ∵ࢆ⏝࠸࡚㸪༸⬊ࡢⓎ⫱ືែ㯤య㏥⾜ㄏ㉳ᚋࡢ༸⬊ࡢᡂ⇍࠾ࡼࡧ༸⮳ࡿ㐣⛬ࡢ༸ ⬊ቨ⾑ὶ㸪ࢫࢸࣟࢻ⃰ᗘ࠾ࡼࡧ㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿᅉᏊ㸪ࡉࡽࡣ༸ᚋᙧ ᡂࡉࢀࡿ㯤యⓎ⫱ࢆ W2DF ẚ㍑ࡋࡓࠋ ➨ 3 ❶࡛ࡣ㸪W1DF ࡢⓎ⫱ࢆไᚚࡍࡿせᅉࢆ᫂ࡽࡍࡿࡓࡵ㸪W1DF ࡢⓎ⫱ࡀ㸪Ⓨ ⫱ᮇ㛫୰ࡢ⾑୰ P4 ⃰ᗘ࠾ࡼࡧྠ୍༸ᕢෆ࡛ࡢ㯤యࡢ᭷↓ࡼࡿ༸ᕢᒁᡤෆ࡛ࡢᙳ㡪ࢆ ཷࡅࡿࡢࢆㄪࡓࠋ ➨ 4 ❶࡛ࡣ㸪Ἢங∵ࢆ⏝࠸㸪W1DF ࡀ⦾Ṫ⏕⌮ࡢࡼ࠺࡞ᙳ㡪ࢆ࠼ࡿࢆ┠ⓗ㸪 ᖍ༸⬊ࡀ㯤యྠഃࡢ༸ᕢᏑᅾࡍࡿሙྜᑐഃᏑᅾࡍࡿሙྜࡢཷ⫾ᛶࢆẚ㍑ࡋ ࡓࠋ 8 ➨㸰❶ ➨ 1 ༸⬊Ἴᖍ༸⬊㸦W1DF㸧ࡢⓎ⫱ືែ㸪༸๓ᚋࡢ ᶵ⬟࠾ࡼࡧ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᛶ㉁ 9 2㸫1㸬ᗎㄽ ࢘ࢩࡢ➨ 1 ༸⬊ἼࡣⓎ࣭༸⤊┤ᚋࡽ༸ᕢෆ࡛Ⓨ⌧ࡍࡿࡇࡀ▱ࡽࢀ࡚࠾ࡾ㸪 ㏻ᖖࡣ㸪㯤యࡀᏑᅾࡍࡿ᮲௳ୗ࡛Ⓨ⫱ࡍࡿࡓࡵ㸪W1DF ࡣⓎ࿘ᮇࡢ 8㹼10 ᪥ࡲ࡛㛢 㙐ࡋ㸪༸⮳ࡿࡇࡣ࡞࠸㸦41㸧 ࠋ࢘ࢩࡢ༸⬊ࡢⓎ⫱ືែ㸪Ⓨ⫱࠾ࡼࡧ㑅ᢤ㛵ࢃࡿෆ ศἪ࣍ࣝࣔࣥືែࢆ᫂ࡽࡍࡿࡇࢆ┠ⓗ㸪ࡇࢀࡲ࡛ W1DF ὀ┠ࡋ࡚ヲ⣽◊✲ ࡀ⾜ࢃࢀ࡚ࡁࡓ㸦9㸪53㸪93㸪103㸧ࡀ㸪W1DF ⮬㌟ࡢᛶ㉁㸪ᶵ⬟࠾ࡼࡧ༸ㄏ㉳ࡢ≉ ᚩࡘ࠸࡚ࡣ༑ศ᳨ウࡉࢀ࡚ࡇ࡞ࡗࡓࠋ➨ 2 ༸⬊ἼࡣⓎ࿘ᮇࡢ 9㹼10 ᪥Ⓨ⌧࣭ Ⓨ⫱ࡋ㸪㯤యࡢ㏥⾜క࠸ᖍ༸⬊ࡣᡂ⇍㸪Ⓨࢆㄏ㉳ࡋ༸⮳ࡿ㸦88㸧 ࠋ➨ 1 ༸⬊Ἴ ࡣ㯤యᙧᡂඹⓎ⫱ࡍࡿࡓࡵ㸪W1DF ࡣ P4 ⃰ᗘࡀప್ࡽᚎࠎୖ᪼ࡍࡿᮇᡂ㛗 ࡍࡿࠋ୍᪉㸪➨ 2 ༸⬊Ἴࡣ㯤యࡀ༑ศᡂ⇍ࡋ࡚ࡽⓎ⫱ࢆ㛤ጞࡍࡿࡓࡵ㸪W2DF ࡣ⾑ ୰ P4 ⃰ᗘࡀ㧗್࡞ᮇᡂ㛗ࡍࡿࠋࡘࡲࡾ㸪୧⪅ࡣ⾑୰ P4 ⃰ᗘ᥎⛣ࡀ␗࡞ࡿ≧ἣୗ࡛ Ⓨ⫱ࡍࡿࡓࡵ㸪ࡑࢀࡒࢀࡢᖍ༸⬊ࡢⓎ⫱ືែ㸪ࢫࢸࣟࢻ⏘⏕⬟㸪༸క࠺ኚ࠾ ࡼࡧ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᙧែࡸᶵ⬟ࡀ␗࡞ࡿྍ⬟ᛶࡀ࠶ࡿࠋ ࡑࡇ࡛㸪ᮏ❶࡛ࡣ㸪W1DF ࡢᛶ㉁࠾ࡼࡧᶵ⬟ࢆ᫂ࡽࡍࡿࡓࡵ㸪㠀Ἢங∵ࢆ⏝࠸࡚㸪 ༸⬊ࡢⓎ⫱ືែ㯤య㏥⾜ㄏ㉳ᚋࡢᡂ⇍㸪༸⮳ࡿ㐣⛬ࡢ༸⬊ቨ⾑ὶ㸪ࢫࢸࣟࢻ⃰ ᗘ࠾ࡼࡧ㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿᅉᏊࡉࡽࡣ༸ᚋᙧᡂࡉࢀࡿ㯤యⓎ⫱ࢆ W2DF ẚ㍑ࡋㄪࡓࠋ 2㸫2㸬ヨ㦂 1㸸༸⬊ἼⓎ⌧ࡽ 7 ᪥┠ࡲ࡛ࡢ W1DF W2DF ࡢⓎ⫱ືែ࠾ࡼࡧ ༸⬊ቨ⾑ὶࡢẚ㍑ 2㸫2㸫1㸬౪ヨື≀ 10 ⮫ᗋⓗᗣ࡛㸪ṇᖖ࡞Ⓨ࿘ᮇࢆ᭷ࡍࡿ࣍ࣝࢫࢱࣥ✀㠀Ἢங∵ 8 㢌㸦ᖹᆒᖺ㱋㸸 5.0 r 1.8 ṓ㸪ᖹᆒsᶆ‽ㄗᕪ㸧ࢆ⏝࠸ࡓࠋ ᑐ㇟∵ࡣᖏᗈ␆⏘Ꮫ␆⏘ࣇ࣮ࣝࢻ⛉Ꮫࢭ ࣥࢱ࣮ෆࡢࣃࢻࢵࢡ࡚㸪ࢥ࣮ࣥࢧ࣮ࣞࢪ㸪ⲡ࠾ࡼࡧ⃰ཌ㣫ᩱࡀΰྜࡉࢀࡓ TMR ࡀ⤥ࡉࢀ㸪⮬⏤㣧Ỉ࡛ࡁࡿ≧ἣ࡛㣫㣴ࡉࢀ࡚࠸ࡓࠋᐇ㦂ෆᐜࡣᖏᗈ␆⏘Ꮫࡢᐇ㦂 ⌮ጤဨ࡚ᢎㄆࡉࢀ࡚࠸ࡿࠋ 2㸫2㸫2㸬ᐇ㦂᪉ἲ 8 㢌ࡢ࢘ࢩࡣ➨ 1 ༸⬊Ἴヨ㦂⩌㸦n㸻4㸧➨ 2 ༸⬊Ἴヨ㦂⩌㸦n㸻4㸧ศࡅࡓࠋࡑࢀ ࡒࢀࡢ⩌ࡣヨ㦂⤊ 2 ᭶ᚋ㸪ࣛࢸࣥ᱁Ꮚἲࢆ⏝࠸࡚ࢡࣟࢫ࣮࢜ࣂ࣮ヨ㦂ࢆ⾜ࡗࡓࠋ ࡑࡢࡓࡵ㸪ࡑࢀࡒࢀࡢ⩌ࡣ 8 㢌ࡢ࢘ࢩࡀ࡚ྵࡲࢀ࡚࠸ࡿ㸦n㸻8㸧 ࠋ ➨ 1 ༸⬊Ἴヨ㦂⩌࡛ࡣ㸪⮬↛Ⓨ㛤ጞᚋ 12 㛫ࡈ㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚༸ ᕢࢆほᐹࡋࡓࠋ༸๓༸⬊ࡢᾘኻࢆ☜ㄆࡋࡓⅬࢆ Day 0 ࡋࡓࠋ༸⬊㑅ᢤࡀ⤊ࡋࡓ ⪃࠼ࡽࢀࡿ Day 3 ࡽ 5 ࡲ࡛ࡢᮇ㛫㸪㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚ᖍ༸⬊ࡢほᐹࢆ ⾜ࡗࡓࠋࡲࡓ Day 6 ࡣ࣮࢝ࣛࢻࣉࣛ⏬ീデ᩿⨨ࢆ⏝࠸࡚ᖍ༸⬊ࡢᖹᆒ┤ᚄ࠾ࡼࡧ ༸⬊ቨ⾑ὶࢆ ᐃࡋࡓࠋࡲࡓ Day 0㹼6 ࡢ㛫㸪1 ᪥ 1 ᅇṇ୰ᑿ㟼⬦ࡼࡾ᥇⾑ࢆ⾜ࡗࡓࠋ ➨ 2 ༸⬊Ἴ⩌࡛ࡣ㸪⮬↛Ⓨ⤊ᚋࡢ༸☜ㄆ᪥ࢆ Day 㸫7 ࡋࡓࠋ➨ 2 ༸⬊ἼⓎ⌧ ࢆㄏ㉳ࡍࡿࡓࡵ㸪Day 㸫2 W1DF ࡢ༸ㄏ㉳ࢆ┠ⓗ GnRH㸦100 μg 㓑㓟ࣇ࢙ࣝࢳ ࣞࣜࣥ㸪➽⫗ෆᢞ 㸹 ࢥࣥࢭ࣮ࣛࣝ㸹 ࣥࢱ࣮࣋ࢵࢺᰴᘧ♫㸪㜰㸪᪥ᮏ㸧ࢆᢞ ࡋࡓࠋGnRH ᢞ 36 㛫ᚋ㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚ W1DF ࡀ༸ࡋ࡚࠸ࡿࡇ ࢆ☜ㄆࡋ㸪ࡑࡢⅬࢆ Day 0 ࡋࡓࠋࡑࡢᚋ㸪➨ 1 ༸⬊Ἴ⩌ྠᵝ༸⬊㑅ᢤࡀ⤊ࡋ ࡓ⪃࠼ࡽࢀࡿ Day 3 ࡽ 5 ࡲ࡛ࡢᮇ㛫㸪㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚ᖍ༸⬊ࡢᖹᆒ ┤ᚄࢆ ᐃࡋࡓࠋࡲࡓ Day 6 ࡣ࣮࢝ࣛࢻࣉࣛ⏬ീデ᩿⨨ࢆ⏝࠸࡚ᖍ༸⬊ࡢᖹᆒ┤ 11 ᚄ࠾ࡼࡧ༸⬊ቨ⾑ὶࡢ ᐃࢆ⾜ࡗࡓࠋࡲࡓ Day 0㹼6 ࡢ㛫㸪1 ᪥ 1 ᅇṇ୰ᑿ㟼⬦ࡼࡾ᥇ ⾑ࢆ⾜ࡗࡓࠋヨ㦂 1 ࡢࣉࣟࢺࢥ࣮ࣝࡢᶍᘧᅗࢆ Fig. 2.1.♧ࡋࡓࠋ 2㸫2㸫3㸬༸⬊ᙧែࡢほᐹ Day 3ࡽ5ࡲ࡛ࡢᖍ༸⬊ࡢᖹᆒ┤ᚄࡣ㸪5.0MHzࡢࣜࢽࣉ࣮ࣟࣈࡀᒓࡋࡓ⤒┤⭠㉸ 㡢Ἴ⏬ീデ᩿⨨㸦HS̺101V㸪ᮏከ㟁Ꮚ㸪ឡ▱㸪᪥ᮏ㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋࣉ࣮ࣟࣈࢆ ᡭᣦࡶ┤⭠ෆᤄධࡋ㸪ᕥྑ୧ഃ༸ᕢࡢ᩿ᒙീࢆほᐹࡋࡓࠋᖍ༸⬊ࡢ༸⬊᩿ᒙ⏬ ീࡀ᭱࡞ࡿࡼ࠺㟼Ṇ⏬ࢆグ㘓ࡋ㸪ࣃࢯࢥࣥୖ㟼Ṇ⏬ࢆ⛣ືࡉࡏ㸪ᖍ༸⬊ࡣ㛗ᚄ ▷ᚄࡢᖹᆒ್ࢆᖹᆒ┤ᚄࡋ㸪ࢹ࣮ࢱࡋ࡚⏝࠸ࡓࠋ Day 6࠾ࡅࡿᖍ༸⬊ࡢ┤ᚄ࠾ࡼࡧ༸⬊ቨ⾑ὶࡣ㸪8.0MHzࡢࣜࢽࣉ࣮ࣟࣈ㸦1739̺ RSࣉ࣮ࣟࣈ㸹GEᶓἙ࣓ࢹ࢝ࣝࢩࢫࢸ࣒㸪ᮾி㸪᪥ᮏ㸧ࢆ᥋⥆ࡋࡓ࣮࢝ࣛࢻࣉࣛ㉸㡢Ἴ ⏬ീデ᩿⨨㸦LOGIQ Book XP㸪GE࣓ࢹ࢝ࣝࢩࢫࢸ࣒㸪ᮾி㸪᪥ᮏ㸧ࢆ⏝࠸࡚࣮࢝ࣛ ࣮ࣔࢻࡼࡾほᐹࢆ⾜ࡗࡓࠋ༸⬊ቨ⾑ὶࡢ᳨ᰝ᪉ἲࡣࣉ࣮ࣟࣈࢆᡭᣦࡶ┤⭠ෆᤄ ධࡋ㸪ᖍ༸⬊ࡢ᩿ᒙീࢆほᐹࡋࡓࠋ༸⬊┤ᚄࡀ࡛᭱ࡘ⾑ὶ㠃✚ࡀ᭱࡞ࡿࡼ࠺࡞ 㟼Ṇ⏬ࢆグ㘓ࡋ㸪ᖹᆒ┤ᚄ㸪༸⬊ቨ⾑ὶ㠃✚㸦BFA㸧࠾ࡼࡧ⾑ὶศᕸྜ㸦BF%㸧ࢆ⟬ฟ ࡋࡓࠋ BFAࡢ⟬ฟࡣ㸪グ㘓ࡋࡓᙳ⏬ീࢆࣃࢯࢥࣥෆྲྀࡾ㎸ࡳ㸪Photoshop 5.5㸦Adobe Systems Inc.㸪San Jose㸪CA㸪USA㸧ࢆ⏝࠸࡚⾑ὶ㡿ᇦࢆⓑ㯮⏬ീኚࡋ㸪Pop Imaging㸦ࢹࢪࢱ ࣭ࣝࣅ࣮ࣥࢢ࣭࢟ࢵࢬ㸪⚄ዉᕝ㸪᪥ᮏ㸧ࢆ⏝࠸࡚⾑ὶ㠃✚ࢆ⟬ฟࡋ㸪BFAࡋ࡚ࢹ࣮ࢱ ࢆ⏝࠸ࡓ㸦Fig. 2.2.㸧ࠋ BF㸣ࡣ㸪ᙳࡋࡓ⏬ീࢆ⏝࠸࡚㸪༸⬊࿘ᑐࡍࡿ⾑ὶศᕸࡢྜࢆホ౯ࡍࡿࡶࡢ࡛࠶ ࡾ㸪༸⬊ቨෆ࡛ࡢ⾑⟶ศᕸࡢ⛬ᗘࢆ♧ࡍᣦᶆࡋ࡚⏝࠸ࡓࠋPhotoshop 5.5ࢆ⏝࠸࡚༸⬊ 12 ࿘⾑ὶศᕸࡀࡳࡽࢀࡿ༸⬊ቨࡢ㛗ࡉࢆ⟬ฟࡋ㸪⾑ὶศᕸࡀࡳࡽࢀࡿ༸⬊ቨࡢ㛗ࡉࢆ༸⬊ ࿘࡛㝖ࡋࡓࡶࡢࢆBF㸣ࡋ࡚ࢹ࣮ࢱࢆ⏝࠸ࡓࠋ 2㸫2㸫4㸬᥇⾑ ヨ㦂ᮇ㛫୰ࡢDay 0ࡽ621G᥇⾑㔪࣊ࣃࣜࣥຍ┿✵᥇⾑⟶㸦࣋ࣀࢪ࢙ࢡࢺϩ┿✵᥇ ⾑⟶㸪TERUMO㸪ᮾி㸧ࢆ⏝࠸࡚ᑿ㟼⬦ࡼࡾ10mlࡢ᥇⾑ࢆ⾜ࡗࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣịỈ ୰࡚ᛴ෭ᚋ㸪4Υ࡛2,000 g㸪20ศ㛫㐲ᚰࡋ㸪⾑₢ศ㞳ᚋ㸪⾑୰P4⃰ᗘ ᐃࡲ࡛㸫30Υ ࡚⤖ಖᏑࡋࡓࠋ 2㸫2㸫5㸬⾑୰P4⃰ᗘࡢ ᐃ ⾑୰P4⃰ᗘࡢ ᐃࡣ2ᢠయἲࡼࡿ㓝⣲ච ᐃἲ㸦EIA㸧ࡼࡾ ᐃࡋࡓࠋEIAࡢᡭ㡰 ࡣ௨๓ࡢሗ࿌ᇶ࡙࠸࡚⾜ࡗࡓ㸦64㸧ࠋ⾑₢ࡼࡾP4ࢆᢳฟࡍࡿࡓࡵ㸪⾑₢200μlࢪ࢚ࢳ ࢚࣮ࣝࢸࣝ㸦㛵ᮾᏛ㸪ᮾி㸪᪥ᮏ㸧1.0mlࢆຍ࠼㸪ᨩᢾᶵ㸦Mix-EVR㸪ࢱࢸࢵࢡ㸪ᇸ ⋢㸪᪥ᮏ㸧ࢆ⏝࠸࡚30ศ㛫࣎ࣝࢸࢡࢫࡋ㸪ࡑࡢᚋᐊ ࡚40ศ㛫㟼⨨ࡋࡓࠋࡑࡢᚋ㸪㸫30Υ ࡢ෭ᗜ⛣ືࡋ㸪24㛫ࡅ࡚⤖ࡉࡏࡓࠋ⤖ᚋ㸪ᾮ┦㒊ศࢆ5mlヨ㦂⟶ࢹ ࢝ࣥࢺࡋ㸪ࢻࣛࣇࢺෆࡢ࢛࣮࢘ࢱ࣮࣭ࣂࢫᾐࡋ㸪40Υࡲ࡛ୖ᪼ࡉࡏ࡞ࡀࡽࢪ࢚ࢳ࢚࣮ࣝ ࢸࣝࢆⓎࡉࡏࡓᚋ㸪50Υࡲ࡛ୖ᪼ࡉࡏ↓⮯࡞ࡿࡲ࡛ⓎࡉࡏࡓࠋⓎᚋ㸪ヨ㦂 ⟶ࢵࢭ࣭ࣂࢵࣇ࣮㸦7.12g㸹NA2HPO4㸪8.5g㸹NaCl㸪1.0g㸹BSA㸪pH 7.2㸹1N-HCL ࡛ㄪᩚ㸧ࢆ200μlຍ࠼㸪20ศ㛫࣎ࣝࢸࢡࢫࡋࡓࠋࡇࡢ ᐃἲࡼࡿP4ࡢᢳฟຠ⋡84㸣࡛࠶ࡗ ࡓࠋEIAࡣᢠ࢘ࢧࢠIgGᢠయ㸦12.5μg/ml㸹⏕Ꮫᕤᴗᰴᘧ♫㸪ᮾி㸪᪥ᮏ㸧࡛⿕ࢃࢀࡓ 96✰ELISAࣉ࣮ࣞࢺ㸦Corning Glass Works㸪Corning㸪New York㸪USA㸧 ࢆ⏝࠸࡚⾜ࡗࡓࠋ ⃰ᗘ ᐃࡣ㸪 ࢫࢱࣥࢲ࣮ࢻ࠾ࡼࡧࢧࣥࣉࣝ15μl100μlࡢP4࣏ࣜࢡ࣮ࣟࢼࣝᢠయ⁐ᾮ 㸦OK-1㸧 㸦1㸸300,000㸧100μlࡢhorseradish peroxidase 㸦1㸸50,000㸧ࢆΰࡋ㸪4Υ࡚20㛫௨ 13 ୖ┞ࡉࡏࡓࠋP4ࡢᶆ‽᭤⥺ࡣ0.05㹼50ng/mlࡢ⠊ᅖ࡛㸪 ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑ ࢀࡒࢀ5.2㸣࠾ࡼࡧ7.4%࡛࠶ࡗࡓࠋ 2㸫2㸫6㸬⤫ィฎ⌮ ⤖ᯝࡣࡍ࡚ᖹᆒ್sᶆ‽ㄗᕪ࡛♧ࡋࡓࠋ⾑୰ P4 ⃰ᗘ᥎⛣ࡢᕪࡢ᳨ᐃࢆ⾜࠺ᙜࡓࡗ ࡚㸪2 ඖ㓄⨨ศᩓศᯒࢆ⾜࠸㸪యࡢኚࡢ᭷ពᛶࢆㄪ㸪స⏝ࡀ᭷ព࡛࠶ࡗࡓሙ ྜ㸪Bartlet ᳨ᐃࢆ⾜࠸ศᩓࡢᆒ୍ᛶࢆ᳨ᐃࡋࡓᚋ㸪Post-hoc ᳨ᐃࡋ࡚ Schefe ࡢከ⩌ẚ ㍑᳨ᐃࢆ⏝࠸ࡓࠋDay 6 ࠾ࡅࡿ༸⬊ቨ⾑ὶࡢẚ㍑ࡣ㸪F ᳨ᐃࢆ⾜࠸ศᩓࡢᆒ୍ᛶࢆ☜ ㄆࡋࡓᚋ㸪Student ࡢ t ᳨ᐃࢆ⏝࠸ࡓࠋ༴㝤⋡ 10㸣ᮍ‶㸦P < 0.1㸧ࢆഴྥ࠶ࡾࡋࠊ༴㝤 ⋡ 5㸣ᮍ‶㸦P < 0.05㸧࡛᭷ពᕪ࠶ࡾࡋࡓࠋ⤫ィฎ⌮ࡣ⤫ィฎ⌮ࢯࣇࢺ࡛࠶ࡿ StatView 㸦Abacus Concepts㸪Inc.㸪Berkeley㸪CA㸪USA㸧ࢆ⏝࠸ࡓࠋ 2㸫3㸬ヨ㦂 2㸸Ⓨ࠾ࡼࡧ༸ㄏ㉳࠾ࡅࡿ W1DF W2DF ࡢⓎ⫱ືែ㸪༸⬊ቨ⾑ὶ ࠾ࡼࡧ⾑୰ E2 ⃰ᗘࡢẚ㍑ 2㸫3㸫1㸬౪ヨື≀ 2㸫2㸬❶ྠᵝࡢື≀ࢆ౪ヨࡋࡓࠋ 2㸫3㸫2㸬ᐇ㦂᪉ἲ ヨ㦂 1 ྠࡌࣉࣟࢺࢥ࣮࡛ࣝ➨ 1 ༸⬊Ἴヨ㦂⩌➨ 2 ༸⬊Ἴヨ㦂⩌ࢆ‽ഛࡋࡓࠋ ࡑࡢᚋ㸪Ⓨ࠾ࡼࡧ༸ㄏ㉳ࢆ⾜࠺ࡇࢆ┠ⓗ㸪Day 6 PGF2α 㸦25 mg ࢺ࣓ࣟࢱ ࣑ࣥࢪࣀࣉࣟࢫࢺ㸪➽⫗ෆᢞ㸪ࣉࣟࢼࣝࢦࣥ F㸪ࢰ࢚ࢸࢫ㸪ᮾி㸪᪥ᮏ㸧ࢆᢞࡋ 㯤య㏥⾜ࢆㄏ㉳ࡋ㸪48 㛫ᚋࡢ Day 8 GnRH ࢆᢞࡋ༸ㄏ㉳ࢆ⾜ࡗࡓࠋDay 6㸪8 14 ࠾ࡼࡧ 9 ࣮࢝ࣛࢻࣉࣛ⏬ീデ᩿⨨ࢆ⏝࠸࡚ᖍ༸⬊ࡢᖹᆒ┤ᚄ࠾ࡼࡧ༸⬊ቨ⾑ὶࡢ ᐃࢆ⾜ࡗࡓࠋࡲࡓ Day 6㹼9 ࡢ㛫㸪1 ᪥ 1 ᅇṇ୰ᑿ㟼⬦ࡼࡾ᥇⾑ࢆ⾜ࡗࡓࠋヨ㦂 2 ࡢ ࣉࣟࢺࢥ࣮ࣝࡢᶍᘧᅗࢆ Fig. 2.5.♧ࡋࡓࠋ 2㸫3㸫3㸬 ༸⬊ᙧែࡢほᐹ Day 6㸪8࠾ࡼࡧ9࠾ࡅࡿᖍ༸⬊ࡢ┤ᚄ࠾ࡼࡧ༸⬊ቨ⾑ὶࡣ㸪8.0MHzࡢࣜࢽࣉ࣮ࣟ ࣈ㸦1739̺RSࣉ࣮ࣟࣈ㸹GEᶓἙ࣓ࢹ࢝ࣝࢩࢫࢸ࣒㸪ᮾி㸪᪥ᮏ㸧ࢆ᥋⥆ࡋࡓ࣮࢝ࣛࢻ ࣉࣛ㉸㡢Ἴ⏬ീデ᩿⨨㸦LOGIQ Book XP㸪GE࣓ࢹ࢝ࣝࢩࢫࢸ࣒㸪ᮾி㸪᪥ᮏ㸧ࢆ⏝ ࠸࡚࣮࣮࢝ࣛࣔࢻࡼࡾほᐹࢆ⾜ࡗࡓࠋ༸⬊ቨ⾑ὶࡢ᳨ᰝ᪉ἲࡣࣉ࣮ࣟࣈࢆᡭᣦࡶ ┤⭠ෆᤄධࡋ㸪ᖍ༸⬊ࡢ᩿ᒙീࢆほᐹࡋࡓࠋ༸⬊┤ᚄࡀ࡛᭱ࡘ⾑ὶ㠃✚ࡀ᭱ ࡞ࡿࡼ࠺࡞㟼Ṇ⏬ࢆグ㘓ࡋ㸪ヨ㦂1ྠᵝࡢ᪉ἲ࡛ᖹᆒ┤ᚄ㸪BFA࠾ࡼࡧBF%ࢆ⟬ฟࡋࡓࠋ 2㸫3㸫4㸬᥇⾑ ヨ㦂ᮇ㛫୰ࡢDay 6㹼921G᥇⾑㔪࣊ࣃࣜࣥຍ┿✵᥇⾑⟶㸦࣋ࣀࢪ࢙ࢡࢺϩ┿✵᥇⾑ ⟶㸪TERUMO㸪ᮾி㸪᪥ᮏ㸧ࢆ⏝࠸࡚ᑿ㟼⬦ࡼࡾ10mlࡢ᥇⾑ࢆ⾜ࡗࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣ ịỈ୰࡚ᛴ෭ᚋ㸪4Υ࡛2,000 g㸪20ศ㛫㐲ᚰࡋ㸪⾑₢ศ㞳ᚋ㸪⾑୰P4⃰ᗘ࠾ࡼࡧ⾑୰E2 ⃰ᗘ ᐃࡲ࡛㸫30Υ࡚⤖ಖᏑࡋࡓࠋ 2㸫3㸫5㸬⾑୰P4࠾ࡼࡧE2⃰ᗘࡢ ᐃ ⾑୰P4⃰ᗘࡣヨ㦂1ྠᵝࡢᡭἲ࡛EIAࡼࡾ ᐃࡋࡓࠋࡇࡢ ᐃἲࡼࡿP4ࡢᢳฟຠ⋡ ࡣ80㸣࡛࠶ࡗࡓࠋ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀࡒࢀ4.7㸣࠾ࡼࡧ6.5㸣࡛࠶ࡗࡓࠋ⾑ ୰E2⃰ᗘࡣDay 8ࡢࡳ ᐃࢆ⾜ࡗࡓࠋ⾑₢ࡼࡾE2ࢆᢳฟࡍࡿࡓࡵ㸪⾑₢2.0mlࢪ࢚ࢳࣝ ࢚࣮ࢸࣝ㸦㛵ᮾᏛ㸪ᮾி㸪᪥ᮏ㸧6.0mlࢆຍ࠼㸪ᨩᢾᶵ㸦DM-301㸪ࢬ࣡ࣥ㸪㜰㸪᪥ 15 ᮏ㸧ࢆ⏝࠸࡚60ศ㛫࣎ࣝࢸࢡࢫࡋ㸪ࡑࡢᚋᐊ ࡚60ศ㛫㟼⨨ࡋࡓࠋࡑࡢᚋ㸫30Υࡢ෭ ᗜ⛣ືࡋ㸪24㛫ࡅ࡚⤖ࡉࡏࡓࠋ⤖ᚋ㸪ᾮ┦㒊ศࢆ10mlヨ㦂⟶ࢹ࢝ࣥࢺ ࡋ㸪ࢻࣛࣇࢺෆࡢ࢛࣮࢘ࢱ࣮࣭ࣂࢫᾐࡋ㸪40Υࡲ࡛ୖ᪼ࡉࡏ࡞ࡀࡽࢪ࢚ࢳ࢚࣮ࣝࢸࣝࢆ Ⓨࡉࡏࡓᚋ㸪50Υࡲ࡛ୖ᪼ࡉࡏ↓⮯࡞ࡿࡲ࡛ⓎࡉࡏࡓࠋⓎᚋ㸪ヨ㦂⟶ ࢵࢭ࣭ࣂࢵࣇ࣮ࢆ200μlຍ࠼㸪20ศ㛫࣎ࣝࢸࢡࢫࡋࡓࠋࡇࡢᡭἲࡼࡾ⾑₢୰ࡢE2ࢆ10 ಸ⃰⦰ࡋࡓࠋࡇࡢ ᐃἲࡼࡿE2ࡢᢳฟຠ⋡ࡣ80㸣࡛࠶ࡗࡓࠋ⃰ᗘ ᐃࡣ㸪ࢫࢱࣥࢲ࣮ࢻ ࠾ࡼࡧࢧࣥࣉࣝ15μl100μlࡢE2࣏ࣜࢡ࣮ࣟࢼࣝᢠయ⁐ᾮ㸦AS-A㸧 㸦1㸸20,000㸧ࢆΰࡋ4Υ ࡚24㛫┞ࡉࡏࡓࠋࡑࡢᚋ㸪100μlࡢhorseradish peroxidase㸦1㸸150,000㸧ࢆΰࡋ㸪4Υ ࡚2㛫┞ࡉࡏࡓࠋE2ࡢᶆ‽᭤⥺ࡣ2㹼2,000 pg/mlࡢ⠊ᅖ࡛㸪 ᐃෆኚື࠾ࡼࡧ ᐃ㛫 ኚືࡣࡑࢀࡒࢀ6.5㸣࠾ࡼࡧ7.6%࡛࠶ࡗࡓࠋ 2㸫3㸫6㸬⤫ィฎ⌮ ⤖ᯝࡣࡍ࡚ᖹᆒ್sᶆ‽ㄗᕪ࡛♧ࡋࡓࠋ⾑୰ P4 ⃰ᗘ㸪ᖍ༸⬊ࡢᖹᆒ┤ᚄ࠾ࡼࡧ༸ ⬊ቨ⾑ὶ᥎⛣ࡢẚ㍑ࡣ㸪2 ඖ㓄⨨ศᩓศᯒࢆ⾜࠸㸪యࡢኚࡢ᭷ពᛶࢆㄪ㸪 స⏝ࡀ᭷ព࡛࠶ࡗࡓࡶࡢᑐࡋ࡚㸪Bartlet ᳨ᐃࢆ⾜࠸ศᩓࡢᆒ୍ᛶࢆ᳨ᐃࡋࡓᚋ㸪 Post-hoc ᳨ᐃࡋ࡚ Schefe ࡢከ⩌ẚ㍑᳨ᐃࢆ⾜ࡗࡓࠋDay 8 ࠾ࡅࡿ⾑୰ E2 ⃰ᗘࡢ᳨ᐃ ࡣ F ᳨ᐃࡼࡾศᩓࡢᆒ୍ᛶࢆ᳨ᐃࡋ㸪Student ࡢ t ᳨ᐃࢆ⾜ࡗࡓࠋP < 0.1 ࢆഴྥ࠶ࡾ ࡋࠊP < 0.05 ࡛᭷ពᕪ࠶ࡾࡋࡓࠋ ⤫ィฎ⌮ࡣ⤫ィฎ⌮ࢯࣇࢺ࡛࠶ࡿ StatView㸦Abacus Concepts㸪Inc.㸪Berkeley㸪CA㸪USA㸧ࢆ⏝࠸ࡓࠋ 2㸫4㸬ヨ㦂 3㸸W1DF W2DF ࠾ࡅࡿ༸๓ࡢ༸⬊ෆࢫࢸࣟࢻ࣍ࣝࣔࣥ⃰ᗘ࠾ࡼࡧ 㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿᅉᏊࡢẚ㍑ 16 2㸫4㸫1㸬౪ヨື≀ ⮫ᗋⓗᗣ࡛㸪ṇᖖ࡞Ⓨ࿘ᮇࢆ᭷ࡍࡿ࣍ࣝࢫࢱࣥ✀㠀Ἢங∵ 7 㢌㸦ᖹᆒᖺ㱋㸸 4.0 r 1.1 ṓ㸪ᖹᆒsᶆ‽ㄗᕪ) ࢆ⏝࠸ࡓࠋ ࢘ࢩࡣᖏᗈ␆⏘Ꮫ␆⏘ࣇ࣮ࣝࢻ⛉Ꮫࢭࣥ ࢱ࣮ෆࡢࣃࢻࢵࢡ࡚㸪ࢥ࣮ࣥࢧ࣮ࣞࢪ㸪ⲡ࠾ࡼࡧ⃰ཌ㣫ᩱࡀΰྜࡉࢀࡓ TMR ࡀ ⤥ࡉࢀ㸪⮬⏤㣧Ỉ࡛ࡁࡿ≧ἣ࡛㣫㣴ࡉࢀ࡚࠸ࡓࠋᐇ㦂ෆᐜࡣᖏᗈ␆⏘Ꮫࡢᐇ㦂⌮ ጤဨ࡚ᢎㄆࡉࢀ࡚࠸ࡿࠋ 2㸫4㸫2 㸬ᐇ㦂᪉ἲ ᐇ㦂ࡢࣉࣟࢺࢥ࣮ࣝࡣヨ㦂 2 ྠᵝ࡛࠶ࡿࠋᮏヨ㦂࡛ࡣ LH ࢧ࣮ࢪ┤๓㸦PGF2α ᢞ 40 㛫ᚋ㸸Day 8㸧࠾ࡼࡧ LH ࢧ࣮ࢪᚋ㸦GnRH ᢞ 24 㛫ᚋ㸸Day 9㸧ࡢᖍ༸⬊ࡢᛶ ㉁ࢆヲ⣽ホ౯ࡍࡿࡓࡵ㸪༸⬊ᾮ࠾ࡼࡧ㢛⢏ᒙ⣽⬊ࢆ᥇ྲྀࡋホ౯ࡋࡓࠋᮏヨ㦂࡛ࡣ௨ୗ 4 ヨ㦂⩌ࢆタࡅࡓ㸪㸯㸧LH ࢧ࣮ࢪ᭚㟢๓➨ 1 ༸⬊Ἴ⩌㸦n㸻7㸧 㸪㸰㸧LH ࢧ࣮ࢪ᭚㟢๓➨ 2 ༸⬊Ἴ⩌㸦n㸻7㸧 㸪㸱㸧LH ࢧ࣮ࢪ᭚㟢ᚋ➨ 1 ༸⬊Ἴ⩌㸦n㸻6㸧 㸪㸲㸧LH ࢧ࣮ࢪ᭚㟢ᚋ ➨ 2 ༸⬊Ἴ⩌㸦n㸻6㸧࡛࠶ࡿࠋ㸯㸧㸰㸧ࡢ⩌ࡣࢡࣟࢫ࣮࢜ࣂ࣮ヨ㦂ࡼࡾ 7 㢌ࡢ࢘ ࢩࡀ࡚ྵࡲࢀ࡚࠸ࡿࠋ㸱㸧㸲㸧ࡢ⩌㛵ࡋ࡚ࡣ 1 㢌ࡢ࢘ࢩ࡛㸪㢛⢏ᒙ⣽⬊ࡢᅇ㔞 ࡀᑡ࡞ࡃࠊࡑࡢ࢘ࢩࢆヨ㦂ࡼࡾ㝖እࡋࡓࡓࡵࠊ6 㢌ࢆ⏝࠸ࡓࢡࣟࢫ࣮࢜ࣂ࣮ヨ㦂࡞ࡗ ࡚࠸ࡿࠋࡲࡓ㸪ࡑࢀࡒࢀࡢ⩌ࡢ࢘ࢩࡢᖍ༸⬊ࡣู࡚ࡢⓎ࿘ᮇ࡛᥇ᮦࢆ⾜࡞ࡗ࡚࠸ ࡿࠋヨ㦂 3 ࡢࣉࣟࢺࢥ࣮ࣝࡢᶍᘧᅗࢆ Fig. 2.9.♧ࡋࡓࠋ 2㸫4㸫3㸬ᖍ༸⬊ࡢ྾ᘬ㢛⢏ᒙ⣽⬊ࡢᅇ ᖍ༸⬊ࡢ྾ᘬࡣ㸪㉸㡢Ἴ⏬ീデ᩿⨨㸦HS-1500㸪ᮏከ㟁Ꮚ㸪ᮾி㸪᪥ᮏ㸧ࢆ⏝࠸࡚ ㉸㡢Ἴ⏬ീほᐹୗ࡛ࡢ⤒⭐༸⬊✸่ࡼࡾ⾜ࡗࡓࠋ㉸㡢Ἴ⏬ീほᐹࡣ㸪7.5MHzࡢ⭐⏝ࣉ ࣮ࣟࣈ㸦HCV-3710MV㸪ᮏከ㟁Ꮚ㸪ᮾி㸪᪥ᮏ㸧ࢆ⏝࠸࡚⾜ࡗࡓࠋࣉ࣮ࣟࣈࡢୖ㒊ࡣ㸪 17 ⣙40cmࡢ✸่⏝ࡢ㔪㸦ࢹࢫ࣏࣮ࢨࣈࣝ᥇༸㔪 A-ࢱࣉ㸪࣑ࢧ࣡་⛉ᕤᴗ㸪Ⲉᇛ㸪᪥ᮏ㸧 ࢆ╔࡛ࡁࡿࡼ࠺࡞ࡗ࡚࠸ࡿࠋ༸⬊྾ᘬࡢ๓‽ഛࡋ࡚㸪⭐ෆ࠾ࡼࡧእ㝜㒊࿘㎶ࢆࡼࡃ Ὑίࡋࡓࠋḟࣉ࣮ࣟࣈࢆ⭐ࡼࡾᤄධࡋ㸪༸ᕢࢆ┤⭠ࡼࡾᤄධࡋࡓᡭ࡛⭐ቨ㉺ࡋᢲࡋᙜ ࡚ࡓࠋ㉸㡢Ἴ⏬ീデ᩿⨨ࡢ⏬㠃ୖࡣ㸪᥇༸㔪ࡢ࢞ࢻࣛࣥࡀ♧ࡉࢀ࡚࠸ࡿࡢ࡛㸪ࡇ ࡢ࢞ࢻࣛࣥୖ྾ᘬࡍࡿ༸⬊ࡀᥥࡉࢀࡿࡼ࠺ࡋࡓࠋ⏬ീࢆほᐹࡋ࡞ࡀࡽ㸪18Gࡢ 㔪ࢆ⭐ቨࢆࡋ࡚༸⬊⭍่ධࡋ㸪༸⬊ᾮ࠾ࡼࡧ㢛⢏ᒙ⣽⬊ࢆ᥇༸㔪㐃⤖ࡉࡏࡓࢩࣜ ࣥࢪࡼࡾ྾ᘬࡋࡓࠋ ༸⬊ᾮࡣ5.0mlࢳ࣮ࣗࣈධࢀ㸪ịỈ୰࡚ᛴ෭ᚋ㸪ᐇ㦂ᐊෆࡢᐇ య㢧ᚤ㙾ୗ࡛㢛⢏ᒙ⣽⬊ࢆᅇࡋࡓࠋ㢛⢏ᒙ⣽⬊ࡣTriPure Isolation Reagent㸦Roche Diagnostics GmbH㸪Mannheim㸪Germany㸧1.0mlࢆຍ࠼࡚⁐ゎࡉࡏ㸪㸫80Υ࡚mRNA ᢳฟࡲ࡛ಖᏑࡋࡓࠋ㢛⢏ᒙ⣽⬊ᅇᚋ㸪༸⬊ᾮࡣ4Υ㸪500 g࡛10ศ㛫㐲ᚰࢆ⾜࠸ୖΎࢆศ 㞳ࡋࡓᚋ㸪EIAࡼࡿP4㸪E2࠾ࡼࡧࣥࢻࣟࢫࢸࣥࢪ࢜ࣥ㸦A4㸧⃰ᗘ ᐃࡲ࡛㸫30Υ࡚ ⤖ಖᏑࡋࡓࠋ 2㸫4㸫4㸬༸⬊ᾮP4㸪E2࠾ࡼࡧA4⃰ᗘࡢ ᐃ ༸⬊ᾮࡼࡾࢫࢸࣟࢻ࣍ࣝࣔࣥࢆᢳฟࡍࡿࡓࡵ㸪⾑₢40μlࢪ࢚ࢳ࢚࣮ࣝࢸࣝ㸦㛵ᮾ Ꮫ㸪ᮾி㸪᪥ᮏ㸧1.0mlࢆຍ࠼㸪ᨩᢾᶵ㸦Mix-EVR㸪ࢱࢸࢵࢡ㸪ᇸ⋢㸪᪥ᮏ㸧ࢆ⏝࠸ ࡚30ศ㛫࣎ࣝࢸࢡࢫࡋ㸪ࡑࡢᚋᐊ ࡚40ศ㛫㟼⨨ࡋࡓࠋࡑࡢᚋ㸫30Υࡢ෭ᗜ࡚⛣ື ࡋ24㛫ࡅ࡚⤖ࡉࡏࡓࠋ⤖ᚋ㸪ᾮ┦㒊ศࢆ5mlヨ㦂⟶ࢹ࢝ࣥࢺࡋ㸪ࢻࣛࣇ ࢺෆࡢ࢛࣮࢘ࢱ࣮࣭ࣂࢫᾐࡋ㸪40Υࡲ࡛ୖ᪼ࡉࡏ࡞ࡀࡽࢪ࢚ࢳ࢚࣮ࣝࢸࣝࢆⓎࡉࡏࡓ ᚋ㸪50Υࡲ࡛ୖ᪼ࡉࡏ↓⮯࡞ࡿࡲ࡛ⓎࡉࡏࡓࠋⓎᚋ㸪ヨ㦂⟶ࢵࢭ࣭ࣂ ࢵࣇ࣮ࢆ200μlຍ࠼㸪20ศ㛫࣎ࣝࢸࢡࢫࡋࡓࠋࡇࡢᡭἲࡼࡾ༸⬊ᾮ୰ࡢࢫࢸࣟࢻ࣍ࣝ ࣔࣥࢆ5ಸᕼ㔘ࡋࡓࠋࡇࡢ ᐃἲࡼࡿP4㸪E2࠾ࡼࡧA4ࡢᢳฟຠ⋡ࡣࡑࢀࡒࢀ82㸣㸪81㸣 ࠾ࡼࡧ85㸣࡛࠶ࡗࡓࠋ P4࠾ࡼࡧE2ࡢ⃰ᗘ ᐃࡣ❶ࡢ2㸫2㸫5.࠾ࡼࡧ2㸫3㸫5.࡛グࡋࡓ࠾ࡾ 18 ࡛࠶ࡿࠋP4ࡢ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀࡒࢀ5.5%࠾ࡼࡧ7.4%㸪E2ࡢ ᐃෆኚື࠾ ࡼࡧ ᐃ㛫ኚືࡣ6.2㸣࠾ࡼࡧ8.6%࡛࠶ࡗࡓࠋA4ࡢ⃰ᗘ ᐃࡣࢫࢱࣥࢲ࣮ࢻ࠾ࡼࡧࢧࣥࣉ ࣝ15μl100μlࡢA4࣏ࣜࢡ࣮ࣟࢼࣝᢠయ⁐ᾮ㸦A-3-CMO-BSA㸪ࢥࢫ࣭ࣔࣂ࢜㸪ᮾி㸪᪥ ᮏ㸧㸦1㸸750,000㸧100μlࡢA-3-CMO-horseradish peroxidase㸦ࢥࢫ࣭ࣔࣂ࢜㸪ᮾி㸪᪥ ᮏ㸧㸦1㸸500,000㸧ࢆΰࡋ4Υ࡚24㛫┞ࡉࡏࡓࠋA4ࡢᶆ‽᭤⥺ࡣ7.8㹼8,000 pg/ml ࡢ⠊ᅖ࡛㸪 ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀࡒࢀ4.3㸣࠾ࡼࡧ5.2%࡛࠶ࡗࡓࠋ 2㸫4㸫5㸬RNAᢳฟࣜࣝࢱ࣒PCR Total RNAࡢᢳฟࡣ௨ୗࡢᡭ㡰࡛⾜ࡗࡓࠋゎࡋࡓࢧࣥࣉࣝࢡ࣒ࣟࣟ࣍ࣝ⁐ᾮࢆ200μl ຍ࠼࡚㸪15⛊㛫࣎ࣝࢸࢡࢫࡋ3ศ㛫ᐊ ࡛㟼⨨ࡋࡓᚋ㸪4Υ㸪36,000 g࡚15ศ㛫㐲ᚰศ㞳 ࢆ⾜ࡗࡓࠋࢡ࣒ࣟࣟ࣍ࣝᾮ┦TriPure Isolation Reagent ⁐ᾮ┦ศ㞳ࡋ࡚࠸ࡿࡇࢆ☜ㄆ ࡋ㸪ࢡ࣒ࣟࣟ࣍ࣝᒙࢆูࡢ1.5mlࢳ࣮ࣗࣈ⛣ࡍࠋࡇࢀ500μlࡢࢯࣉࣟࣃࣀ࣮ࣝ⁐ᾮ 㸦3-Propanol㸧ࢆຍ࠼㸪15⛊㛫࣎ࣝࢸࢡࢫࡋ㸪10ศ㛫ᐊ ࡚㟼⨨ࡋࡓᚋ㸪4Υ㸪36,000 g ࡚15ศ㛫㐲ᚰศ㞳ࢆ⾜ࡗࡓࠋRNAࡢ࣌ࣞࢵࢺࡢᏑᅾࢆ☜ㄆࡋࡓᚋ㸪ࢯࣉࣟࣃࣀ࣮ࣝ⁐ ᾮࢆ㝖ཤࡋ㸪500μlࡢ80㸣࢚ࢱࣀ࣮ࣝࢆຍ࠼㸪Ὑίࡋ㸪4Υ㸪36,000 g࡚5ศ㛫㐲ᚰศ㞳ࢆ ⾜ࡗࡓࠋ࢚ࢱࣀ࣮ࣝࢆ㝖ཤᚋ㸪ᐊ ࡛5ศ㛫࣌ࣞࢵࢺࢆ⇱ࡉࡏ㸪6μlࡢࢪ࢚ࢳࣝࣆ࣮ࣟ࢝ ࣎ࢿ࣮ࢺ㸦DEPC㸧ฎ⌮Ỉࢆຍ࠼ࡓࠋ ㏫㌿ࡣSuperscript VIRO Reaction Mix 㸦Invitrogen.㸪Carlsbad㸪CA㸪USA㸧ࢆ⏝࠸ࡓࠋ ࡲࡎ㸪Nano-Drop 2000㸦Thermo scientific㸪Wilmington㸪DE㸪USA㸧ࢆ⏝࠸࡚RNA⃰ᗘࢆ⟬ ฟࡋࡓࠋḟ㸪RQ1 RNase-Free DNase (Promega㸪Madison㸪WI㸪USA) ࢆ⏝࠸࡚DNaseฎ ⌮ࢆ⾜ࡗࡓࠋRNAࡀ1μgྵࡲࢀࡿRNA⁐ᾮ㸪1μlࡢDNase 10×Reaction Buffer࠾ࡼࡧ2μlࡢ RNase-Free DNaseࢆΰࡋ㸪37Υ࡛30ศ㛫࣮ࣥ࢟ࣗ࣋ࢺࡋࡓࠋࡑࡢᚋ㸪1μlࡢDNase Stop Solutionࢆຍ࠼65Υ࡛10ศ㛫࣮ࣥ࢟ࣗ࣋ࢺࡋࡓࠋDNaseฎ⌮ࡋࡓࢧࣥࣉࣝ5×VILO 19 Reaction Mixࢆ4μl㸪10×Super Script Enzymeࢆ2μlຍ࠼㸪⥲㔞ࡀ20μl࡞ࡿࡼ࠺DEPCฎ⌮ Ỉࢆຍ࠼ࡓᚋ㸪25Υ࡛10ศ㛫㸪42Υ࡛60ศ㛫࠾ࡼࡧ85Υ࡛5ศ㛫࣮ࣥ࢟ࣗ࣋ࢺࡋ㸪cDNA ࢆసᡂࡋࡓࠋ 2㸫4㸫6㸬ࣜࣝࢱ࣒PCRࡼࡿmRNAⓎ⌧㔞ࡢ ᐃ ࣜࣝࢱ࣒PCRࡣTli RNaseH Plus㸦࢟ࢤࣥ㸪ᮾி㸪᪥ᮏ㸧ࢆ⏝ࡋ㸪Roter-Gene Q 㸦࢟ ࢤࣥ㸪ᮾி㸪᪥ᮏ㸧 ࢆ⏝࠸࡚⾜ࡗࡓࠋ7.5μlࡢSYBR Green Master Mix㸪5.75μlࡢDEPCฎ ⌮Ỉ㸪ForwardReverseࡢࣉ࣐࣮ࣛࢆྛ0.375μlຍ࠼㸪᭱ᚋ1.0μlࡢࢧࣥࣉࣝࢆຍ࠼࡚ ᛂࡉࡏࡓࠋ ᐃࡍࡿmRNAࡣ㸪LHཷᐜయ㸦LHr㸧㸪࣐ࣟࢱ࣮ࢮ㸦CYP19A1㸧㸪ࢩࢡࣟ࢜ ࢟ࢩࢤࢼ࣮ࢮ̺2㸦COX-2㸧㸪PGE2ྜᡂ㓝⣲㸦PGES㸧㸪PGF2αྜᡂ㓝⣲㸦PGFS㸧㸪ㄏᑟᆺ ୍㓟❅⣲ྜᡂ㓝⣲㸦iNOS㸧㸪⾑⟶ෆ⓶ᆺ୍㓟❅⣲ྜᡂ㓝⣲㸦eNOS㸧㸪⾑⟶ෆ⓶ቑṪ ᅉᏊ㸦VEGF㸧120㸪VEGF164㸪⥺⥔ⱆ⣽⬊ቑṪᅉᏊ㸦FGF㸧-2㸪Steroidogenic acute regulatory protein㸦StAR㸧㸪ࢳࢺࢡ࣮࣒ࣟP450ഃ㙐ษ᩿㓝⣲㸦cytochrome P450 cholesterol side-chain cleavage enzymes㸹 P450-scc㸧㸪3β-Ỉ㓟ᇶ⬺Ỉ⣲㓝⣲㸦3β-hydroxy steroid dehydrogenase㸹 3β-HSD㸧࠾ࡼࡧࢢࣜࢭࣝࣝࢹࣄࢻ3ࣜࣥ㓟⬺Ỉ⣲㓝⣲㸦glyceraldehyde-3-phosphate dehydrogenase㸹GAPDH㸧࡛࠶ࡿࠋGAPDHࡣෆ㒊ᶆ‽㑇ఏᏊࡋ࡚⏝࠸㸪ࡑࡢⓎ⌧ࡢ ྜ࡛ồࡵࡓࠋࣉ࣐࣮ࣛࡣPrimer-3ࢆ⏝࠸࡚సᡂࡋࡓࠋࣜࣝࢱ࣒PCRࡢᛂ᮲௳ࡣ94Υ ࡛10ศ㛫ฎ⌮ࡋࡓᚋ㸪94Υ࡛30⛊㸪ྛࠎࡢࢽ࣮ࣜࣥࢢ ᗘ࡛30⛊࠾ࡼࡧ72Υ࡛30⛊ࡢࢧ ࢡࣝࢆ40ᅇ⾜ࡗࡓࠋࡑࢀࡒࢀࡢmRNAࡢࣉ࣐࣮ࣛࡢࢩ࣮ࢡ࢚ࣥࢫࢆ♧ࡋࡓ⾲ࢆTable 2.3.♧ࡋࡓࠋLHࢧ࣮ࢪ᭚㟢๓ࡢ㢛⢏ᒙ⣽⬊ᑐࡋ࡚ࡣLHr࠾ࡼࡧCYP19A1 mRNAⓎ⌧㔞 ࢆ ᐃࡋࡓࠋLHࢧ࣮ࢪ᭚㟢ᚋࡢ㢛⢏ᒙ⣽⬊ᑐࡋ࡚ࡣCOX-2㸪PGES㸪PGFS㸪iNOS㸪eNOS㸪 VEGF120㸪VEGF164㸪FGF-2㸪StAR㸪P450-scc࠾ࡼࡧ3β-HSD mRNAⓎ⌧㔞ࢆ ᐃࡋࡓࠋ 20 2㸫4㸫7㸬⤫ィฎ⌮ ⤖ᯝࡣࡍ࡚ᖹᆒ್sᶆ‽ㄗᕪ࡛♧ࡋࡓࠋ༸⬊ᾮ୰ࡢᛶࢫࢸࣟࢻ࣍ࣝࣔࣥ⃰ᗘ࠾ࡼ ࡧ㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡍࡿ mRNA Ⓨ⌧㔞ࡢẚ㍑㛵ࡋ࡚ࡣ㸪ࡲࡎ F ᳨ᐃࡼࡾศᩓࡢᆒ୍ ᛶࢆ᳨ᐃࡋ㸪➼ศᩓࡀㄆࡵࡽࢀࡓሙྜ㸪Student ࡢ t ᳨ᐃࢆ⾜࠸㸪➼ศᩓࡀㄆࡵࡽࢀ࡞ ࡗࡓሙྜࡣ Mann–Whitney ࡢ U ᳨ᐃࢆ⾜ࡗࡓࠋP < 0.1 ࢆഴྥ࠶ࡾࡋࠊP < 0.05 ࡛᭷ព ᕪ࠶ࡾࡋࡓࠋ⤫ィฎ⌮ࡣ⤫ィฎ⌮ࢯࣇࢺ࡛࠶ࡿ StatView㸦Abacus Concepts㸪Inc.㸪 Berkeley㸪CA㸪USA㸧ࢆ⏝࠸ࡓࠋ 2㸫5㸬ヨ㦂 4㸸W1DF W2DF ࡢ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᙧែ㸪㯤య⾑ὶ࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ࡢẚ㍑ 2㸫5㸫1㸬౪ヨື≀ ⮫ᗋⓗᗣ࡛㸪ṇᖖ࡞Ⓨ࿘ᮇࢆ᭷ࡍࡿ࣍ࣝࢫࢱࣥ✀㠀Ἢங∵ 6 㢌㸦ᖹᆒᖺ㱋㸸 4.2 r 2.1 ṓ㸪ᖹᆒsᶆ‽ㄗᕪ) ࢆ⏝࠸ࡓࠋ ࢘ࢩࡣᖏᗈ␆⏘Ꮫ␆⏘ࣇ࣮ࣝࢻ⛉Ꮫࢭࣥ ࢱ࣮ෆࡢࣃࢻࢵࢡ࡚㸪ࢥ࣮ࣥࢧ࣮ࣞࢪ㸪ⲡ࠾ࡼࡧ⃰ཌ㣫ᩱࡀΰྜࡉࢀࡓ TMR ࡀ ⤥ࡉࢀ㸪⮬⏤㣧Ỉ࡛ࡁࡿ≧ἣ࡛㣫㣴ࡉࢀ࡚࠸ࡓࠋᐇ㦂ෆᐜࡣᖏᗈ␆⏘Ꮫࡢᐇ㦂⌮ ጤဨ࡚ᢎㄆࡉࢀ࡚࠸ࡿࠋ 2㸫5㸫2 㸬ᐇ㦂᪉ἲ 6 㢌ࡢ࢘ࢩࡣ W1DF ༸ᚋ㯤య㸦W1CL㸹n㸻3㸧 W2DF ༸ᚋ㯤య㸦W2CL㸹n㸻3㸧 ศࡅࡓࠋࡑࢀࡒࢀࡢ⩌ࡣヨ㦂⤊ 2 ᭶ᚋ㸪ࣛࢸࣥ᱁Ꮚἲࢆ⏝࠸࡚ࢡࣟࢫ࣮࢜ࣂ࣮ ヨ㦂ࢆ⾜ࡗࡓࠋࡑࡢࡓࡵ㸪ࡑࢀࡒࢀࡢ⩌ࡣ 6 㢌ࡢ࢘ࢩࡀ࡚ྵࡲࢀ࡚࠸ࡿ㸦n㸻6㸧 ࠋ 21 ヨ㦂ࡢࣉࣟࢺࢥ࣮ࣝࡣヨ㦂 2 ྠᵝ࡛࠶ࡿࡀ㸪ᮏヨ㦂࡛ࡣᖍ༸⬊ࢆ྾ᘬࡏࡎ㸪༸ ࡉࡏ㸪ࡑࡢᚋᙧᡂࡉࢀࡿ㯤యࢆほᐹࡋࡓࠋ W1DF W2DF ࡶ㸪Day 8 GnRH ࢆᢞࡋ㸪ࡑࡢ 24 㛫࠾ࡼࡧ 36 㛫ᚋ㉸ 㡢Ἴ⏬ീデ᩿⨨࡛༸☜ㄆࢆ⾜࠸㸪24 㛫ᚋ༸ࡏࡎ㸪36 㛫ᚋࡲ࡛༸ࡀ☜ㄆ ࡉࢀࡓⅬࢆ Day 10 ࡋࡓࠋDay 9 ࠾ࡅࡿᖍ༸⬊ࡢᖹᆒ┤ᚄ࠾ࡼࡧ༸⬊ቨ⾑ὶ㸪Day 12㸪15㸪18 ࠾ࡼࡧ 21 ࠾ࡅࡿ㯤యࡢ㯤య᩿㠃✚࠾ࡼࡧ㯤య⾑ὶࢆ࣮࢝ࣛࢻࣉࣛ㉸㡢Ἴ ⏬ീデ᩿⨨ࢆ⏝࠸࡚ ᐃࡋࡓࠋ ࡲࡓ Day 10㹼21 ࡢ㛫㸪1 ᪥ 1 ᅇṇ୰ᑿ㟼⬦ࡼࡾ᥇⾑ࢆ⾜ࡗࡓࠋヨ㦂 4 ࡢࣉࣟࢺࢥ࣮ ࣝࡢᶍᘧᅗࢆ Fig. 2.15.♧ࡋࡓࠋ 2㸫5㸫3㸬༸⬊࠾ࡼࡧ㯤యᙧែࡢほᐹ Day 9࠾ࡅࡿᖍ༸⬊ࡢᖹᆒ┤ᚄ࠾ࡼࡧ༸⬊ቨ⾑ὶ㸪Day 12㸪15㸪18࠾ࡼࡧ21࠾ࡅ ࡿ㯤యࡢ᩿㠃✚࠾ࡼࡧ㯤య⾑ὶࡣ㸪8.0MHzࡢࣜࢽࣉ࣮ࣟࣈ㸦1739̺RSࣉ࣮ࣟࣈ㸹GEᶓ Ἑ࣓ࢹ࢝ࣝࢩࢫࢸ࣒㸪ᮾி㸪᪥ᮏ㸧ࢆ᥋⥆ࡋࡓ࣮࢝ࣛࢻࣉࣛ㉸㡢Ἴ⏬ീデ᩿⨨㸦LOGIQ Book XP㸪GE࣓ࢹ࢝ࣝࢩࢫࢸ࣒㸪ᮾி㸪᪥ᮏ㸧ࢆ⏝࠸࡚࣮࣮࢝ࣛࣔࢻࡼࡾほᐹࢆ⾜ࡗ ࡓࠋ༸⬊ࡢホ౯ࡣヨ㦂1ྠᵝࡢ᪉ἲ࡛⾜࠸㸪༸⬊ቨ⾑ὶࡢホ౯ࡣBF㸣ࢆ⏝࠸ࡓࠋ㯤య ࡢ᩿㠃✚ࡣ㸪㯤యࡢ᩿ᒙ⏬ീࡀ᭱࡞ࡿࡼ࠺㟼Ṇ⏬ࢆグ㘓ࡋ㸪ࣃࢯࢥࣥୖ㟼Ṇ⏬ࢆ⛣ ືࡉࡏ㸪㯤యࡢ㛗ᚄ▷ᚄࡢᖹᆒ್ࡽᖹᆒ┤ᚄࢆ⟬ฟࡋ㸪㸦ᖹᆒ┤ᚄ㸭2㸧2×3.14ᙜ࡚ ࡣࡵ㯤య᩿㠃✚ࢆ⟬ฟࡋ㸪ࢹ࣮ࢱ⏝࠸ࡓࠋ 㯤య⾑ὶࡢ᳨ᰝ᪉ἲࡣ㸪㯤యࡢ᩿ᒙീࢆほᐹࡋ㸪㯤య᩿㠃✚ࡀ࡛᭱ࡘ⾑ὶ㠃✚ࡀ᭱ ࡞ࡿࡼ࠺࡞㟼Ṇ⏬ࢆグ㘓ࡋ㸪㯤య⾑ὶ㠃✚ࢆ⟬ฟࡋࡓࠋ 㯤య⾑ὶ㠃✚ࡢ⟬ฟࡣ㸪グ㘓ࡋࡓᙳ⏬ീࢆࣃࢯࢥࣥෆྲྀࡾ㎸ࡳ㸪Photoshop 5.5 㸦Adobe Systems Inc.㸪San Jose㸪CA㸪USA㸧ࢆ⏝࠸࡚⾑ὶ㡿ᇦࢆⓑ㯮⏬ീኚࡋ㸪Pop 22 Imaging㸦ࢹࢪࢱ࣭ࣝࣅ࣮ࣥࢢ࣭࢟ࢵࢬ㸪⚄ዉᕝ㸧ࢆ⏝࠸࡚⾑ὶ㠃✚ࢆ⟬ฟࡋ㸪㯤య⾑ ὶ㠃✚ࡋ࡚ࢹ࣮ࢱࢆ⏝࠸ࡓࠋ 2㸫5㸫4㸬᥇⾑ ヨ㦂ᮇ㛫୰ࡢDay 10㹼2121G᥇⾑㔪࣊ࣃࣜࣥຍ┿✵᥇⾑⟶㸦࣋ࣀࢪ࢙ࢡࢺϩ┿✵᥇ ⾑⟶㸪TERUMO㸪ᮾி㸧ࢆ⏝࠸࡚ᑿ㟼⬦ࡼࡾ10mlࡢ᥇⾑ࢆ⾜ࡗࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣịỈ ୰࡚ᛴ෭ᚋ㸪4Υ࡛2,000 g㸪20ศ㛫㐲ᚰࡋ㸪⾑₢ศ㞳ᚋ㸪⾑୰P4⃰ᗘ ᐃࡲ࡛㸫30Υ ࡚⤖ಖᏑࡋࡓࠋ 2㸫5㸫5㸬⾑୰P4⃰ᗘࡢ ᐃ ⾑୰P4⃰ᗘࡢ ᐃࡣヨ㦂1ྠᵝࡢ᪉ἲ࡛⾜ࡗࡓࠋ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀ ࡒࢀ6.4㸣࠾ࡼࡧ8.6%࡛࠶ࡗࡓࠋ 2㸫5㸫6㸬⤫ィฎ⌮ ⤖ᯝࡣࡍ࡚ᖹᆒ್sᶆ‽ㄗᕪ࡛♧ࡋࡓࠋ㯤య᩿㠃✚㸪㯤య⾑ὶ㠃✚࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ࢆẚ㍑ࡍࡿᙜࡓࡗ࡚ࡣ㸪2 ඖ㓄⨨ศᩓศᯒࢆ⾜࠸㸪యࡢኚࡢ᭷ពᛶࢆㄪ 㸪స⏝ࡀ᭷ព࡛࠶ࡗࡓሙྜ㸪Bartlet ᳨ᐃࢆ⾜࠸ศᩓࡢᆒ୍ᛶࢆ᳨ᐃࡋࡓᚋ㸪Post-hoc ᳨ᐃࡋ࡚ Schefe ࡢከ⩌ẚ㍑᳨ᐃࢆ⏝࠸ࡓࠋ༸๓༸⬊ࡢᖹᆒ┤ᚄࢆẚ㍑ࡍࡿࡣ㸪ࡲ ࡎ F ᳨ᐃࡼࡾศᩓࡢᆒ୍ᛶࢆ☜ㄆࡋࡓᚋ㸪Student ࡢ t ᳨ᐃࢆ⾜ࡗࡓࠋP < 0.1 ࢆഴྥ࠶ ࡾࡋࠊP < 0.05 ࡛᭷ពᕪ࠶ࡾࡋࡓࠋ⤫ィฎ⌮ࡣ⤫ィฎ⌮ࢯࣇࢺ࡛࠶ࡿ StatView 㸦Abacus Concepts㸪Inc.㸪Berkeley㸪CA㸪USA㸧ࢆ⏝࠸ࡓࠋ 2㸫6㸬⤖ᯝ 23 2㸫6㸫1㸬ヨ㦂1ࡢ⤖ᯝ ⾑୰P4⃰ᗘ᥎⛣ࢆFig. 2.3.♧ࡋࡓࠋヨ㦂ᮇ㛫୰࠾࠸࡚W2⩌࡛W1⩌ẚ㍑ࡋ࡚᭷ព 㧗ࡃ᥎⛣ࡋࡓࠋ ᖍ༸⬊ࡢᖹᆒ┤ᚄࡢ᥎⛣ࢆFig. 2.4.♧ࡋࡓࠋW1DFࡢᖹᆒ┤ᚄࡣヨ㦂ᮇ㛫୰㸪W2DF ẚ㍑ࡋ࡚ࡁࡃⓎ⫱ࡋ㸪≉Day 6᭷ពࡁࡗࡓࠋࡲࡓ㸪W1DF⩌ෆ࡛ࡣ㸪Day 3 ࠾ࡼࡧ4ࡽDay 6ࡅ࡚᭷ពࡁࡃⓎ⫱ࡋࡓࡀ㸪W2DF⩌ෆ࡛ࡣ㸪Day 3ࡽDay 6 ࡅ࡚ࡢࡳ᭷ពࡁࡃ࡞ࡗࡓࠋ Day 6࠾ࡅࡿBFA࠾ࡼࡧBF㸣ࢆTable 2.1.♧ࡋࡓࠋBFA࠾ࡼࡧBF%ࡣW1DF࡛㧗࠸ഴ ྥࢆ♧ࡋࡓࠋ 2㸫6㸫2㸬ヨ㦂2ࡢ⤖ᯝ ᖍ༸⬊ࡢᖹᆒ┤ᚄࡢ᥎⛣ࢆFig. 2.6.♧ࡋࡓࠋᖹᆒ┤ᚄࡢ᥎⛣ࡣ㸪W1DFࡀW2DFẚ ㍑ࡋ࡚ヨ㦂ᮇ㛫୰ࢆ㏻ࡋ࡚᭷ព㧗ࡃ᥎⛣ࡋࡓࠋヨ㦂ᮇ㛫୰ࡢ⾑୰P4⃰ᗘ᥎⛣ࢆFig. 2.7. ♧ࡋࡓࠋDay 6࠾ࡅࡿ⾑୰P4⃰ᗘࡣW2DF࡛W1DFẚ㍑ࡋ࡚᭷ព㧗ࡗࡓࡀ㸪ࡑࡢ ᚋࡢ⾑୰P4⃰ᗘ᥎⛣୧⩌㛫࡛ᕪࡣ࡞ࡃ㸪PGF2Dᢞᚋᛴ⃭పୗࡋࡓࠋ ヨ㦂ᮇ㛫୰ࡢBFA࠾ࡼࡧBF%ࡢ᥎⛣ࢆFig. 2.8.♧ࡋࡓࠋBFAࡣ㸪W1DF࠾࠸࡚Day 9 ࡛W2DFẚ㍑ࡋ࡚᭷ព㧗ࡗࡓࡀ㸪Day 6࠾ࡼࡧ8࡛ࡣ୧⩌㛫࡛ᕪࡣ࡞ࡗࡓࠋࡲࡓ㸪 BFAࡢ᥎⛣ࢆ⩌ෆ࡛ẚ㍑ࡋࡓࡇࢁW1DF࠾ࡼࡧW2DFࡶDay 6ࡽ9ࡅ࡚᭷ព ቑຍࡋࡓࠋࡲࡓ㸪W1DF࠾࠸࡚ࡢࡳDay 8ࡽ9ࡅ࡚᭷ពቑຍࡋࡓࡀ㸪W2DF࡛ࡣ ᭷ព࡞ቑຍࡣࡳࡽࢀ࡞ࡗࡓࠋBF㸣ࡣ㸪W1DF࠾࠸࡚Day 9࡛W2DFẚ㍑ࡋ࡚᭷ព㧗 ࡗࡓࡀ㸪Day 6࠾ࡼࡧ8࡛ࡣ୧⩌㛫࡛ᕪࡣ࡞ࡗࡓࠋBF㸣᥎⛣ࢆ⩌ෆ࡛ẚ㍑ࡋࡓࡇࢁ 24 W1DF࡛ࡣDay 6ࡽ9࠾ࡼࡧDay 8ࡽ9ࡅ࡚᭷ពቑຍࡋࡓࡀ㸪W2DF࡛ࡣ᭷ព࡞ቑ ຍࡣࡳࡽࢀ࡞ࡗࡓࠋ Day 8࠾ࡅࡿ⾑୰E2⃰ᗘࢆTable 2.2.♧ࡋࡓࠋ⾑୰E2⃰ᗘࡣW1DF࡛᭷ព㧗ࡗࡓࠋ 2㸫6㸫3㸬ヨ㦂3ࡢ⤖ᯝ LHࢧ࣮ࢪ᭚㟢๓࠾ࡅࡿ༸⬊ᾮ୰E2࠾ࡼࡧA4⃰ᗘࡣ㸪W1DF࡛᭷ព㧗ࡗࡓࡀ㸪P4 ⃰ᗘᕪࡣ࡞ࡗࡓ㸦Table 2.4.㸧ࠋLHࢧ࣮ࢪ᭚㟢ᚋ࠾ࡅࡿ㸪༸⬊ᾮ୰P4࠾ࡼࡧE2⃰ᗘ ᕪࡣ࡞ࡗࡓ㸦Table 2.5.㸧ࠋ LHrCYP19A1 mRNAⓎ⌧㔞ࡢ⤖ᯝࢆFig. 2.10.♧ࡋࡓࠋLHrࡣW1DF࡛W2DFẚ㍑ࡋ ࡚᭷ព㧗ࡗࡓࡀ㸪CYP19AⓎ⌧㔞ᕪࡣ࡞ࡗࡓࠋCOX-2㸪PGES࠾ࡼࡧPGFS mRNA Ⓨ⌧㔞୧⩌㛫࡛ᕪࡣ࡞ࡗࡓ㸦Fig. 2.11㸧ࠋiNOS࠾ࡼࡧeNOS mRNAⓎ⌧㔞ࢆFig. 2.12. ♧ࡋࡓࠋiNOS ᕪࡣ࡞ࡗࡓࡀ㸪eNOSࡣW1DF࡛㧗࠸ഴྥࢆ♧ࡋࡓࠋVEGF120㸪 VEGF164࠾ࡼࡧFGF-2 mRNAⓎ⌧㔞ࡢ⤖ᯝࢆFig. 2.13.♧ࡋࡓࠋW1DF࠾࠸࡚㸪VEGF120 ࠾ࡼࡧFGF-2࡛㧗࠸ഴྥࢆ♧ࡋ㸪VEGF164ࡣ᭷ព㧗ࡗࡓࠋStAR㸪P450-scc࠾ࡼࡧ3β-HSD mRNAⓎ⌧㔞ࡢ⤖ᯝࢆFig. 2.14.♧ࡋࡓࠋW1DF࠾࠸࡚StAR࡛㧗࠸ഴྥࢆ♧ࡋ㸪P450-scc ࠾ࡼࡧ3β-HSD mRNAࡣ᭷ព㧗ࡗࡓࠋ 2㸫6㸫4㸬ヨ㦂4ࡢ⤖ᯝ Day 9࠾ࡅࡿW1DFW2DFࡢᖹᆒ┤ᚄBF㸣ࡢ⤖ᯝࢆTable 2.6.♧ࡋࡓࠋW1DF࠾ ࠸࡚ᖹᆒ┤ᚄ࠾ࡼࡧBF%ࡣࡁࡗࡓࠋ㯤య᩿㠃✚᥎⛣ࢆFig. 2.16.♧ࡋࡓࠋ㯤య᩿㠃✚ ࡣDay 15㸪18࠾ࡼࡧ21࡛W1CL࡛W2CLẚ㍑ࡋ࡚᭷ព㧗ࡗࡓࠋ 25 㯤య⾑ὶ㠃✚᥎⛣ࢆFig. 2.17.♧ࡋࡓࠋ㯤య⾑ὶ㠃✚ࡣDay 12࠾ࡼࡧ15࡛W1CL࡛W2CL ẚ㍑ࡋ࡚᭷ព㧗ࡗࡓࠋ ⾑୰P4⃰ᗘ᥎⛣ࢆFig .2.18.♧ࡋࡓࠋ⾑୰P4⃰ᗘࡣヨ㦂ᮇ㛫ࢆ㏻ࡌ࡚W1CL࡛W2CL ẚ㍑ࡋ࡚᭷ព㧗ࡗࡓࠋ 2㸫7㸬⪃ᐹ ᮏ❶࡛ࡢヨ㦂 1 ࠾ࡼࡧ 2 ࡽ㸪W1DF ࡣ W2DF ẚ㍑ࡋ࡚㸪ᖍ༸⬊ࡢᖹᆒ┤ᚄ㸪༸ ⬊ቨ⾑ὶ㸪LH ࢧ࣮ࢪ᭚㟢┤๓ࡢ⾑୰ E2 ⃰ᗘࡀ㧗ࡗࡓࠋࡲࡓ㸪ヨ㦂 3 ࡽ㸪༸⬊ᾮ E2 ⃰ᗘ࠾ࡼࡧ༸⬊ᾮ A4 ⃰ᗘ㸪㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿ LHr㸪⾑⟶᪂⏕ᅉᏊ㸪ࢫࢸࣟࢻ ྜᡂ㓝⣲ࡢ mRNA Ⓨ⌧㔞ࡀ W1DF ࡛㧗ࡗࡓࠋヨ㦂 4 ࡽ㸪W1DF ༸ᚋᙧᡂࡉࢀ ࡿ㯤యࡢ᩿㠃✚㸪⾑ὶ࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ࡀ㧗࠸ࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡇࢀࡽࡢ⤖ ᯝࡽ㸪W1DF ࡣ W2DF ẚ㍑ࡋ࡚༸⬊ࡋ࡚ࡼࡾᶵ⬟ⓗάຊࡀ㧗࠸ࡇࡀ⪃࠼ࡽࢀ ࡿࠋࡲࡓ㸪ࡇࢀࡽࡢᕪ␗ࡀ⏕ࡌࡿཎᅉࡋ࡚㸪W1DF Ⓨ⫱ᮇ㛫୰ࡢప࠸⾑୰ P4 ⃰ᗘక ࠺㧗࠸ LH ࣃࣝࢫ㢖ᗘࡼࡿ༸⬊⣽⬊ࡢ่⃭ࡀ᥎ ࡉࢀࡓࠋ ヨ㦂 1 ࡼࡾ㸪༸⬊ἼⓎ⌧㛤ጞࡽ 7 ᪥┠ࡲ࡛ࡢᖍ༸⬊ࡢⓎ⫱ࢆẚ㍑ࡍࡿ㸪༸⬊㑅 ᢤ㛤ጞᚋࡽ㸪Ⓨ⫱⋡ࡸᖍ༸⬊ࡢᖹᆒ┤ᚄࡣ W1DF ࡛ࡁࡃ࡞ࡿࡇࡀ♧ࡉࢀࡓࠋࡲ ࡓ㸪W1DF ࠾ࡼࡧ W2DF ࡑࢀࡒࢀࡢⓎ⫱ᮇ㛫୰ࡢ⾑୰ P4 ⃰ᗘࢆẚ㍑ࡍࡿ㸪Day 0 ࡽ Day 6 ࡅ࡚ W1DF ࡛᭷ពపࡃ᥎⛣ࡍࡿࡇࡀ᫂ࡽࡉࢀࡓࠋ㐣ཤࡢ◊✲࠾࠸ ࡚㸪㯤యᙧᡂᮇࡣ㯤య㛤ⰼᮇẚ㍑ࡋ࡚㸪LH ࣃࣝࢫ㢖ᗘࡀ㧗ࡃ࡞ࡿࡇࡀሗ࿌ࡉࢀ࡚ ࠸ࡿ㸦73㸧 ࠋࡲࡓ㸪⭐ෆ␃⨨ᆺ P4 〇ࢆ⏝࠸࡚㸪⾑୰ P4 ⃰ᗘࢆ㧗ࡃࡍࡿ LH ࣃࣝࢫ㢖 ᗘࡣపୗࡍࡿ୍᪉࡛㸪⾑୰ P4 ⃰ᗘࢆపࡃ⥔ᣢࡍࡿ LH ࣃࣝࢫ㢖ᗘࡣୖ᪼ࡍࡿࡇࡀ▱ ࡽࢀ࡚࠸ࡿ㸦15㸪52㸧 ࠋࡲࡓ㸪Adams et al.㸦5㸧ࡣ㸪⮬↛Ⓨᚋࡢ༸☜ㄆ⭐ෆ␃⨨ ᆺ P4 〇ࢆᤄධࡋ㸪ࡑࡢᚋࡢ༸ᕢෆ࡛Ⓨ⫱ࡍࡿ➨ 1 ༸⬊Ἴࡢ༸⬊Ⓨ⫱ືែࢆほᐹࡋࡓ 26 ࡇࢁ㸪ᤄධࡋ࡞ࡗࡓ∵ẚᖍ༸⬊ࡢ᭱㐩ࡋࡓᖹᆒ┤ᚄࡀᑠࡉ࠸ࡇࢆሗ࿌ࡋ ࡚࠸ࡿࠋ༸⬊Ἴࡢ୰࡛༸⬊ࡀᖍ༸⬊ࡋ࡚㑅ᢤࡉࢀ㸪ࡑࡢᚋⓎ⫱ࢆ⥅⥆ࡍࡿࡓࡵࡣ㸪 㢛⢏ᒙ⣽⬊࡛ LHr ࡢⓎ⌧ࡀᚲ㡲࡛࠶ࡿ㸦13㸧 ࠋヨ㦂 1 ࡛ࡳࡽࢀࡓ㸪W1DF ࡢ W2DF ẚ ࡚ᖹᆒ┤ᚄࡀࡁࡃ᥎⛣ࡋࡓࡇࡽ㸪W1DF ࡢⓎ⫱ᮇ㛫୰ࡢప࠸⾑୰ P4 ⃰ᗘ᥎⛣ ㉳ᅉࡍࡿ㧗࠸ LH ࣃࣝࢫ㢖ᗘࡼࡾᖍ༸⬊ࡢⓎ⫱ࡀಁࡉࢀࡓ᥎ ࡉࢀࡿࠋࡉࡽ㸪 ヨ㦂 1 ࡛ࡣ Day 6 ༸⬊ቨ⾑ὶࢆほᐹࡋࡓࡇࢁ㸪W1DF ࡣ W2DF ẚ㍑ࡋ࡚㧗࠸ഴྥ ࢆ♧ࡋࡓࠋ༸⬊ࡢⓎ⫱㐣⛬࠾࠸࡚㸪༸⬊ቨ࡛ࡢ⾑⟶ᙧᡂࡣ㔜せ࡛࠶ࡿࡇࡀ▱ࡽࢀ࡚ ࠸ࡿ㸦95㸧 ࠋ㐣ཤࡢ◊✲࠾࠸࡚㸪㑅ᢤᚋࡢᖍ༸⬊࡛ࡢ༸⬊ቨ⾑ὶࡀቑຍࡍࡿࡇࡀሗ ࿌ࡉࢀ࡚࠸ࡿ㸦42㸧 ࠋࡲࡓ㸪㛢㙐๓ᚋࡢ༸⬊ቨ⾑ὶࢆẚ㍑ࡍࡿ㛢㙐ᚋ༸⬊ቨ⾑ὶࡀほ ᐹࡉࢀ࡞ࡃ࡞ࡿ࠸࠺ሗ࿌ࡶ࠶ࡿ㸦2㸧 ࠋࡇࢀࡽࡢࡇࡽ㸪࣮࢝ࣛࢻࣉࣛࢆ⏝࠸ࡓ༸⬊ ቨ⾑ὶࡢホ౯ࡣ༸⬊ࡢάຊࡸᶵ⬟ࢆホ౯ࡍࡿࡢ᭷ຠ࡞ᣦᶆ࡞ࡿ⪃࠼ࡽࢀ㸪ᮏ◊✲ ࡛ࡳࡽࢀࡓW1DFW2DF ࠾ࡅࡿ༸⬊ቨ⾑ὶࡢ㐪࠸ࡣ༸⬊ࡢᶵ⬟ࡢᕪࢆ♧ࡋ࡚࠸ࡿ ⪃࠼ࡽࢀࡿࠋ ᮏ◊✲ࡢ⤖ᯝ࠾࠸࡚㸪W1DF ࡛ࡣ W2DF ẚⓎ⫱⋡㸪ᖹᆒ┤ᚄ࠾ࡼ ࡧ༸⬊ቨ⾑ὶࡀ㧗ࡗࡓࡇࡽ㸪ᶵ⬟ᛶࡀ㧗࠸༸⬊࡛࠶ࡿࡇࡀ᥎ ࡉࢀࡓࠋ ヨ㦂 2 ࡛ࡣ PGF2α ࠾ࡼࡧ GnRH ࢆ⏝࠸࡚㸪㯤య㏥⾜ㄏ㉳ᚋࡢ༸⬊ࡢᡂ⇍࠾ࡼࡧ༸ ⮳ࡿ㐣⛬ࡢ W1DF W2DF ࡢᛶ㉁ࢆㄪ㸪ẚ㍑ࡋࡓࠋᖍ༸⬊ࡢᖹᆒ┤ᚄࡣ Day 6 ࡽ Day 9 ࡅ࡚ᮇ㛫ࢆ㏻ࡋ࡚ W1DF ࡛ࡁࡃ᥎⛣ࡋࡓࠋ༸⬊ቨ⾑ὶࡢኚࢆẚ㍑ࡋࡓ ࡇࢁ㸪W1DF ࠾࠸࡚ GnRH ᢞ๓ᚋ࡛᭷ព࡞ቑຍࡀほᐹࡉࢀࡓࡀ㸪W2DF ࠾࠸࡚ࡣ ᭷ព࡞ቑຍࡣほᐹࡉࢀࡎ㸪Day 9 ࠾࠸࡚ W1DF ࡛༸⬊ቨ⾑ὶࡀ W2DF ẚ᭷ព ࡁࡗࡓࠋ ࢘ࢩ࡛ࡣ LH ࢧ࣮ࢪࡽ࠾ࡼࡑ 27 㛫ᚋ༸ࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ 㸦24㸧 ࠋ ࡲࡓ࢘ࢩ࡛ࡣ㸪༸┤๓㸦LH ࢧ࣮ࢪ᭚㟢࠾ࡼࡑ 19㹼21 㛫ᚋ㸧ࡢ༸⬊ቨ⾑ὶࡀቑຍࡍ ࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ 㸦85㸧 ࠋ༸⬊ࡀ LH ࢧ࣮ࢪ᭚㟢ࡉࢀࡿ㸪༸⬊ෆ࡛ PG 㢮㸪 ࣄࢫࢱ࣑ࣥ㸪ࢥࣛࢤࢼ࣮ࢮ㸪࢟ࢽࣥ࠾ࡼࡧࣉࣛࢫ࣑ࣀ࣮ࢤࣥࢡࢳ࣮࣋ࢱ࡞ࡢ⏘⏕ࡀ 27 ஹ㐍ࡋ༸⬊⾑⟶⥙࡛ࡢ㏱㐣ᛶஹ㐍ࡸ༸⬊ቨࡢ⼥ゎ㸪⳼ⷧࡀ㉳ࡇࡿࡇ࡛༸ࡀ㐍⾜ࡋ㸪 ࡑࡢᛂࡣ⅖⌧㇟㢮ఝࡋ࡚࠸ࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ㸦25㸧 ࠋࡑࡢࡓࡵ㸪LH ࢧ࣮ࢪ᭚ 㟢ᚋቑຍࡍࡿ༸⬊ቨ⾑ὶࡣ㸪ࡇࢀࡽࡢ⅖ᛶኚ㉳ᅉࡍࡿ⾑⟶ᶵ⬟ࡢኚࡼࡿ ⪃࠼ࡽࢀࡿࠋࡋࡋ࡞ࡀࡽ㸪ᮏヨ㦂࡛ࡣ GnRH ᢞ 24 㛫ᚋ࠾ࡅࡿ༸⬊ቨ⾑ὶࡢ᭷ ព࡞ቑຍࡣ W1DF ࡢࡳ࡛ࡋほᐹࡉࢀ࡞ࡗࡓࠋࡲࡓ㸪BF㸣࠾࠸࡚ࡶ Day 9 ࠾࠸࡚ W1DF ࡛㧗࠸ࡇࡽ㸪W1DF ࡢ⾑ὶࡢቑຍ࠾ࡼࡧ㧗್ࡢཎᅉࡋ࡚㸪༸⬊ቨෆ࡛ࡢ⾑ ⟶ࡢศᕸࡀࡼࡾᗈ⠊ᅖ࡛࠶ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋࡲࡓ㸪LH ࢧ࣮ࢪᑐࡍࡿᛂᛶࡀ 㧗࠸ࡇ࡛༸⬊ቨ࡛ࡢ⅖ᛶኚࡀࡼࡾࡁࡃ࡞ࡾ㸪⾑⟶ࡢ⾑ᾮ㑏ὶ㔞ࡀቑຍࡋ࡚࠸ ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋᮍ⤒⏘∵ࢆ⏝࠸࡚Ⓨㄏ㉳ࢆ⾜࡞ࡗࡓヨ㦂࠾࠸࡚㸪GnRH ᢞ 26 㛫ᚋࡢ༸⬊ቨ⾑ὶࡑࡢᐇࡋࡓேᕤᤵ⢭ᚋࡢཷ⫾ࡢ᭷↓ࢆẚ㍑ࡋࡓࡇ ࢁ㸪ዷፎࡋࡓ∵࡛༸⬊ቨ⾑ὶࡀ᭷ព㧗࠸ࡇࢆሗ࿌ࡋ࡚࠸ࡿ㸦84㸧 ࠋࡇࡢ⌧㇟ࡣ㸪࣐࢘ ࢆ⏝࠸ࡓヨ㦂࠾࠸࡚ࡶྠᵝࡢ⤖ᯝࡀᚓࡽࢀ࡚࠸ࡿ㸦87㸧 ࠋࡲࡓ㸪Ⓨࢆㄏ㉳ࡋ GnRH ᢞ 26 㛫ᚋࡢ༸๓༸⬊ࡽᅇࡋࡓ༸Ꮚᑐࡋ࡚㸪యእཷ⢭ࢆ⾜࠸ࡑࡢᚋࡢ⬇Ⓨ⫱ ࢆẚ㍑ࡋࡓ◊✲࡛ࡣ㸪༸Ꮚᅇࡢ༸๓༸⬊ࡢ༸⬊ቨ⾑ὶࡀࡁ࠸ሙྜ㸪⬇Ⓨ⫱ࡀⰋ ዲ࡛࠶ࡿࡇࡶ᫂ࡽࡉࢀ࡚࠸ࡿ㸦86㸧 ࠋࡇࢀࡽࡢࡇࡽ㸪༸๓༸⬊ࡢ༸⬊ቨ⾑ὶ ࡣ㸪ࡑࡢ༸⬊ࡢάຊࡢ⛬ᗘࢆ♧ࡍࡇࡀ࡛ࡁࡿ⪃࠼ࡽࢀࡿࠋࡲࡓ㸪ᮏ◊✲ࡽ㸪GnRH ᢞࡢ Day 8 ࠾ࡅࡿ⾑୰ E2 ⃰ᗘࡣ W1DF ࡛㧗࠸ࡇࡀ♧ࡉࢀࡓࠋ㐣ཤࡢᐃேᕤᤵ ⢭ࢆ⏝࠸ࡓヨ㦂࠾࠸࡚㸪GnRH ᢞࡢ⾑୰ E2 ⃰ᗘࡑࡢᚋࡢཷ⫾ྍ⬟ᛶࡢ㛫ࡣ ṇࡢ┦㛵㛵ಀࡀᏑᅾࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ㸦14㸧 ࠋࡲࡓ㸪ᐃேᕤᤵ⢭Ⓨᚩೃࢆ ♧ࡋ࡚࠸ࡓ࢘ࢩࡣ㸪Ⓨᚩೃࢆ♧ࡋ࡚࠸࡞ࡗࡓ࢘ࢩẚ㍑ࡋ࡚ཷ⫾⋡ࡀ㧗࠸ࡇࡀሗ ࿌㸦70㸧ࡉࢀ࡚࠾ࡾ㸪༸๓ࡢ⾑୰ E2 ⃰ᗘࡣཷ⫾ᛶࡸ༸⬊ࡢάຊࢆ♧ࡍᣦᶆ࡞ࡿࠋࡇ ࢀࡽࡢࡇࡽ㸪 W1DFࡣ༸⬊ቨ⾑ὶࡀࡁࡃ㸪 ༸┤๓ࡢ⾑୰E2 ⃰ᗘࡀ㧗࠸ࡇࡽ㸪 W2DF ẚάຊࡀ㧗࠸ࡇࡀ᥎ ࡉࢀࡿࠋ 28 ヨ㦂 3 ࡛ࡣ㸪ヨ㦂 2 ࡛ᚓࡽࢀࡓ㸪W1DF ࡢᛶ㉁ࢆࡉࡽヲ⣽ẚ㍑ࡍࡿࡓࡵ㸪LH ࢧ ࣮ࢪ᭚㟢๓ᚋࡢ༸⬊ᾮᛶࢫࢸࣟࢻ࣍ࣝࣔࣥࡸ㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿ mRNA Ⓨ⌧ 㔞ࢆẚ㍑ࡋࡓࠋ LH ࢧ࣮ࢪ᭚㟢๓ࡢᡂ⇍༸⬊ෆ࡛㸪ࢫࢸࣟࢻ࣍ࣝࣔࣥྜᡂ㐣⛬ࡣ㸪༸⬊ቨࡢෆ༸⬊ ⭷⣽⬊࠾࠸࡚㸪ࢫࢸࣟࢻ࣍ࣝࣔࣥࡢᇶ㉁࡞ࡿࢥࣞࢫࢸ࣮ࣟࣝࡀ⣽⬊ෆྲྀࡾ㎸ࡲ ࢀࡿࡇࡽጞࡲࡾ㸪ࡇࡢࢥࣞࢫࢸ࣮ࣟࣝࡣ StAR ࡼࡾ࣑ࢺࢥࣥࢻࣜෆྲྀࡾ㎸ࡲ ࢀ㸪⥆࠸࡚࣑ࢺࢥࣥࢻࣜෆ⭷ୖᏑᅾࡍࡿ P450-scc ࡼࡾࢥࣞࢫࢸ࣮ࣟࣝࡽࣉࣞࢢ ࢿࣀࣟࣥኚࡉࢀ㸪ࡑࡢᚋ㠃ᑠ⬊యୖᏑᅾࡋ࡚࠸ࡿ 3β-HSD ࡼࡾࣉࣞࢢࢿࣀࣟ ࣥࡽ P4 ኚࡉࢀࡿ㸦22㸧 ࠋP4 ࡣ C17 Ỉ㓟㓝⣲άᛶࡼࡾ 17α̺ࣄࢻࣟ࢟ࢩࣉࣟ ࢪ࢙ࢫࢸࣟࣥኚࡉࢀ㸪C17-20 ഃ㙐ษ᩿㓝⣲άᛶࡼࡾ A4 ኚࡉࢀࡿ㸦31㸪59㸧 ࠋ ࡉࡽ A4 ࡣ༸⬊⭷⣽⬊ࡽ㢛⢏ᒙ⣽⬊⛣⾜ࡋࡑࡇ࡛࣐ࣟࢱ࣮ࢮ㸦CYP19A1㸧ࡼ ࡾ E2 ኚࡉࢀࡿ㸦12㸧ࡇ࡛༸⬊ෆ࡛ࢫࢸࣟࢻࡀ⏘⏕ࡉࢀ࡚࠸ࡿࠋᮏヨ㦂࠾࠸࡚㸪 ༸⬊ᾮ୰ E2 ࠾ࡼࡧ A4 ⃰ᗘࡣ W1DF ࡛㧗ࡗࡓࡀ㸪CYP19A1 mRNA Ⓨ⌧㔞࠾࠸࡚ࡣ ୧⩌ᕪࡣ࡞ࡗࡓࠋ㐣ཤ⾜ࢃࢀࡓ◊✲࠾࠸࡚㸪༸⬊ෆ࡛ࡢ E2 ⏘⏕ࡢไ㝈せᅉࡣ㸪 ࣐ࣟࢱ࣮ࢮάᛶ࡛ࡣ࡞ࡃ㸪ෆ༸⬊⭷࡛⏘⏕ࡉࢀࡿ A4 ࡼࡿࡇࢁࡀࡁ࠸࠸࠺ሗ࿌ ࡀ࠶ࡿ㸦9㸧 ࠋࡇࢀࡽࡢࡇࡽ㸪W1DF ࠾ࡅࡿ༸⬊ᾮ E2 ⃰ᗘࡢ㧗್ࡣ W1DF ࡢෆ༸ ⬊⭷࡛ࡢ㧗࠸ A4 ⏘⏕⬟ࡼࡿࡶࡢ࡛࠶ࡿ᥎ ࡉࢀࡿࠋLH ࢧ࣮ࢪ᭚㟢๓ࡢ㢛⢏ᒙ⣽⬊ ࡛Ⓨ⌧ࡋ࡚࠸ࡿ LHr mRNA Ⓨ⌧㔞ࢆẚ㍑ࡍࡿ㸪W1DF ࡀ㧗࠸ࡇࡀ♧ࡉࢀࡓࠋ࢘ࢩࡢ 㢛⢏ᒙ⣽⬊࠾࠸࡚㸪LH ࡣ LHr mRNA Ⓨ⌧㔞ࢆቑຍࡉࡏࡿࡇࡀ▱ࡽࢀ࡚࠾ࡾ㸦56㸧 㸪 ヨ㦂 1 ࠾࠸࡚ W1DF Ⓨ⫱ᮇ㛫୰ࡢ⾑୰ P4 ⃰ᗘࡀపࡗࡓࡇࡽ㸪 W1DF Ⓨ⫱ᮇ㛫୰ ࡢ㧗࠸LHࣃࣝࢫ㢖ᗘࡀW1DF㢛⢏ᒙ⣽⬊୰࠾ࡅࡿLHrⓎ⌧㔞ࢆ㧗ࡵࡓ⪃࠼ࡽࢀࡿࠋ ༸⬊ࡀ LH ࢧ࣮ࢪᛂࡋ࡚༸࡛ࡁࡿࡼ࠺࡞ࡿࡓࡵࡣ㸪㢛⢏ᒙ⣽⬊࠾ࡅࡿ LHr Ⓨ⌧ࡀᚲ㡲࡛࠶ࡿࡇࡽ㸪W1DF ࡢ㢛⢏ᒙ⣽⬊ࡣ LH ࢧ࣮ࢪࢆཷࡅ࡚༸ྥ࠺ 29 ᛂᛶࡀࡼࡾ㧗࠸ࡇࡀ᥎ ࡉࢀ㸪ヨ㦂 2 ࠾ࡅࡿ Day 9 ࡛ࡢ W1DF ࡢ༸⬊ቨ⾑ὶࡢ㧗್ ࡣ㸪W1DF ࡢ LH ࢧ࣮ࢪᑐࡍࡿ༸๓ࡢ༸⬊ቨ⾑ὶቑຍ࠾ࡅࡿᛂᛶࡢ㧗ࡉࢆᫎ ࡋ࡚࠸ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡿࠋ ༸⬊ࡀ LH ࢧ࣮ࢪ᭚㟢ࡉࢀࡿ㸪༸⬊ࢆᵓᡂࡍࡿ⣽⬊࡛࠶ࡿ㢛⢏ᒙ⣽⬊ࡸ༸⬊⭷⣽ ⬊ࡣ㯤య⣽⬊ኚࡍࡿࡇ࡛㸪ࢫࢸࣟࢻྜᡂࡣ E2 ྜᡂࡽ P4 ྜᡂᛴ㏿⛣ ⾜ࡋ࡚࠸ࡃ㸦89㸧 ࠋࡲࡓ㸪LH ࢧ࣮ࢪ᭚㟢ᚋࡢ༸⬊ᾮ୰࠾࠸࡚㸪PG 㢮ࡢ⃰ᗘࡀᛴ⃭ ୖ᪼ࡋ࡚࠸ࡃ㸦1㸪3㸧 ࠋPG ࡣᒁᡤ࠾ࡅࡿ⾑⟶సືᛶస⏝㸦21㸪98㸧ࡸ⾑⟶㏱㐣ᛶస⏝ 㸦26㸧ࢆᣢࡘࡇࡀ▱ࡽࢀ࡚࠾ࡾ㸪༸┤๓ࡢ༸⬊య✚ቑຍ㸦61㸧ࡑࡢᚋࡢ༸⬊◚ 㸦26㸧㛵ࢃࡿࡉࢀ࡚࠸ࡿࠋࡉࡽ㸪༸┤๓࠾ࡅࡿ PGE2 ྜᡂࢆᢚไࡍࡿ㸪༸ ⬊ࡢ◚ࢆᢚไࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ㸦71㸧 ࠋ࢘ࢩ࠾࠸࡚㏥⾜㛤ጞࡢ㯤య࠾ࡅࡿ ⾑ὶࡢቑຍࡣ⾑⟶ᣑᙇᅉᏊ࡛࠶ࡿ୍㓟❅⣲㸦NO㸧ࡀ㛵ࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠾ࡾ 㸦83㸧 㸪࣐࢘ࢫ࠾࠸࡚༸⬊ෆ࡛ࡶⓎ⌧ࡀ☜ㄆࡉࢀ࡚࠸ࡿ㸦104㸧 ࠋVEGF ࡸ FGF ࡣ༸⬊ Ⓨ⫱㸦82㸧ࡸ㯤యᙧᡂ㸦105㸧࠾ࡅࡿ⾑⟶᪂⏕㔜せ࡞ᙺࢆᯝࡓࡍࡇࡀ▱ࡽࢀ࡚࠾ ࡾ㸪ࡇࢀࡽࡢᅉᏊࡢⓎ⌧㔞ࢆẚ㍑ࡍࡿࡇ࡛ LH ࢧ࣮ࢪ๓ᚋ༸⮳ࡿ㐣⛬࠾ࡅࡿ ༸⬊ᶵ⬟ࢆホ౯࡛ࡁࡿ⪃࠼ࡓࠋ ᮏヨ㦂࡛ࡣ㸪COX-2㸪PGES ࠾ࡼࡧ PGFS mRNA Ⓨ⌧㔞᭷ព࡞ᕪࡀ࡞ࡗࡓࡇ ࡽ㸪༸㐣⛬ᚲ㡲࡞ࡇࢀࡽࡢᅉᏊࡢⓎ⌧㔞ཬࡰࡍ㸪LH ࢧ࣮ࢪᑐࡍࡿᛂᛶࡀ W1DF W2DF ࡢ㛫࡛ᕪ␗ࡣ࡞࠸ࡇࢆ♧၀ࡋ࡚࠸ࡿ⪃࠼ࡽࢀࡿࠋࡘࡲࡾ㸪ࡇࢀࡽ ࡢᅉᏊࡢⓎ⌧㔞ࡢ㐪࠸ࡀ㸪༸⬊ቨ⾑ὶࡢᕪ␗ࢆࡶࡓࡽࡋࡓཎᅉ࡛࠶ࡿࡣ⪃࠼ࡽࢀ࡞࠸ࠋ ୍᪉࡛㸪eNOS mRNA Ⓨ⌧㔞ࡣ W1DF ࡛㧗࠸ഴྥࢆ♧ࡋࡓࡇࡽ㸪༸⬊ࡢ⾑⟶ᣑᙇస ⏝ W1DF W2DF 㛫࡛ᕪࡀ⏕ࡌ࡚࠸ࡿྍ⬟ᛶࡶ⪃࠼ࡽࢀࡿࠋࡋࡋ࡞ࡀࡽ㸪༸⬊ቨࡢ ⾑⟶⥙ࡣෆ༸⬊⭷ศᕸࡋ࡚࠾ࡾ㸪㢛⢏ᒙ⣽⬊ࡣ⾑⟶ࡀධࡋ࡚࠸࡞࠸㸦29㸧ࡇ ࡽ㸪ෆ༸⬊⭷࡛ࡢ⾑⟶⥙ࡢศᕸࡸෆ༸⬊⭷⣽⬊࡛ࡢ iNOS ࠾ࡼࡧ eNOS ࡞ࡢ⾑⟶సື 30 ᛶᅉᏊࡢⓎ⌧㔞ࢆẚ㍑ࡍࡿࡇ࡛㸪༸⬊ቨ⾑ὶࡢᕪ␗ࢆࡶࡓࡽࡋࡓཎᅉࡘ࠸࡚㏣ཬࡋ ࡚࠸ࡃᚲせࡀ࠶ࡿ⪃࠼ࡿࠋ P4 ⏘⏕㛵ࢃࡿࢫࢸࣟࢻྜᡂ㓝⣲࡛࠶ࡿ StAR㸪P450-scc ࠾ࡼࡧ 3β-HSD mRNA Ⓨ⌧ 㔞ࢆ W1DF W2DF 㛫࡛ẚ㍑ࡋࡓࡇࢁ㸪P450-scc ࠾ࡼࡧ 3β-HSD mRNA ࡀ W1DF ࡛ 㧗ࡃⓎ⌧ࡋ࡚࠸ࡓࠋࡲࡓ㸪⾑⟶ᙧᡂᅉᏊ࡛࠶ࡿ VEGF120㸪164 ࠾ࡼࡧ FGF-2 mRNA Ⓨ⌧ 㔞࠾࠸࡚ࡶ㸪VEGF164 mRNA Ⓨ⌧㔞ࡀ W1DF ࡛㧗ࡃⓎ⌧ࡋ࡚࠸ࡓࠋ㢛⢏ᒙ⣽⬊ᑐ ࡋ࡚㸪LH ࡣ VEGF㸦45㸧࠾ࡼࡧ P450-scc 㸦106㸧ࡢⓎ⌧ࢆୖ᪼ࡉࡏࡿሗ࿌ࡉࢀ࡚࠾ࡾ㸪 W1DF ࡢ LH ࢧ࣮ࢪ᭚㟢๓ࡢ㢛⢏ᒙ⣽⬊࡛ LHr mRNA ࡢⓎ⌧ࡀ㧗ࡗࡓࡇࡽ㸪ࡇࢀ ࡽࡢᅉᏊࡢⓎ⌧㔞ࡀୖ᪼ࡋࡓ᥎ ࡉࢀࡿࠋࡇࡢࡼ࠺ࢫࢸࣟࢻྜᡂ㓝⣲࠾ࡼࡧ⾑⟶ ᪂⏕ᅉᏊࡢⓎ⌧ࡀ W1DF ࡛㧗ࡃ࡞ࡗ࡚࠸ࡓࡇࡽ㸪༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᙧែࡸ ᶵ⬟ࡀ W1CL ࠾࠸࡚㧗࠸ࡇࡀ᥎ ࡉࢀࡓࠋ ヨ㦂 4 ࠾࠸࡚㸪W1DF ༸ᚋᙧᡂࡉࢀࡿ W1CL ࡢ㯤య᩿㠃✚㸪㯤య⾑ὶ㠃✚࠾ࡼ ࡧ⾑୰ P4 ⃰ᗘࡣ㧗ࡗࡓࠋࡇࢀࡲ࡛㸪༸๓༸⬊ VEGF ࡢࣥࢱࢦࢽࢫࢺࢆᢞࡋࡓ ሙྜ㸪ࡑࡢᚋᙧᡂࡉࢀࡿ㯤యࡢᙧែࡸᶵ⬟ࢆపୗࡉࡏࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸦46㸧 ࠋ ࡲࡓ㸪FGFR1 ᑐࡍࡿࣥࣄࣅࢱ࣮ࢆᇵ㣴ࡋࡓ㯤య⣽⬊ῧຍࡍࡿ⾑⟶ෆ⓶⣽⬊ࡢࢿ ࢵࢺ࣮࣡ࢡࡸ⾑⟶ഃᯞࢆⴭࡋࡃῶᑡࡉࡏࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ㸦101㸧 ࠋࡇࢀࡽࡢࡇ ࡽ㸪W1DF ࠾ࡅࡿ⾑⟶ᙧᡂᅉᏊࡢ㧗࠸Ⓨ⌧ࡼࡾ㸪㯤యᙧᡂึᮇࡢ⾑⟶ᙧᡂࡢ⛬ᗘࡀ W1CL ࡛ࡁࡃ࡞ࡗࡓ⪃࠼ࡽࢀࡿࠋࡇࢀࡲ࡛㸪༸๓༸⬊ࡢ┤ᚄࡀࡁ࠸ሙྜ㸪ࡑࡢ ᚋᙧᡂࡉࢀࡿ㯤యࡢ┤ᚄࡶࡁࡃ࡞ࡿ࠸࠺ሗ࿌ࡀ࠶ࡿ㸦97㸧 ࠋᮏヨ㦂࠾࠸࡚ࡶ༸ ๓༸⬊ࡣ W1DF ࡛ࡁࡃ㸪ࡑࡢᚋࡢ㯤య᩿㠃✚ࡀࡁ࠸ࡇࡽ㸪㐣ཤࡢሗ࿌୍⮴ࡍ ࡿࠋࡉࡽ㸪W1CL ࡛ࡣ㯤యᙧᡂึᮇ࡛ࡢ⾑⟶ᙧᡂࡢ⛬ᗘࡀࡁࡗࡓࡇࡽ㸪⾑ᾮ 㑏ὶ㔞ࡀቑຍࡋ㸪㯤యࡢⓎ⫱ࡀࡉࡽಁࡉࢀ࡚ࡁࡃⓎ⫱ࡋࡓ⪃࠼ࡽࢀࡿࠋ㐣ཤࡢሗ ࿌࠾࠸࡚㸪㯤యࡢ┤ᚄ⾑୰ P4 ⃰ᗘࡣṇࡢ┦㛵㛵ಀࡀᏑᅾࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿ 31 㸦58㸧 ࠋᮏヨ㦂࠾࠸࡚ࡶ㸪⾑୰ P4 ⃰ᗘࡀ㧗ࡃ᥎⛣ࡋࡓཎᅉ㸪W1CL ࡢࢧࢬࡀࡁ ࠸ࡇࡶ⪃࠼ࡽࢀࡿࡀ㸪㯤యᙧᡂึᮇ⾑ὶ㔞ࡀከ࠸ࡇ࡛㯤య⣽⬊࡛ࡢ P4 ⏘⏕ࡢஹ㐍 ࡸ㸪⏘⏕ࡉࢀࡓ P4 ࢆࡼࡾຠ⋡Ⰻࡃ㌟ᚠ⎔ᨺฟ࡛ࡁࡓࡇࡀ㧗࠸⾑୰ P4 ⃰ᗘࢆࡶࡓ ࡽࡋࡓࡢ࡛ࡣ࡞࠸᥎ ࡉࢀࡿࠋࡲࡓ㸪ヨ㦂 3 ࡽ P4 ྜᡂ㓝⣲ࡢ mRNA Ⓨ⌧㔞ࡀ㧗 ࡗࡓࡇࡽ㸪㯤య⣽⬊࡛ࡢࢫࢸࣟࢻྜᡂ⬟ࡢᶵ⬟ࡢ㧗࠸ࡇࡶせᅉࡢ୍ࡘ࡞ࡿ ⪃࠼ࡽࢀࡿࠋࡇࢀࡽࡢࡇࡽ㸪W1DF ࡣ༸⬊ࡢᶵ⬟ࡔࡅ࡛࡞ࡃ㸪ࡑࡢᚋᙧᡂࡉࢀ ࡿ㯤యࡢάຊࡶ㧗࠸ࡇࡀ♧ࡉࢀࡓࠋࡲࡓ㸪㯤యᶵ⬟ࡣ༸┤๓ࡢ༸⬊ᶵ⬟ࡁࡃᙳ 㡪ࢆཷࡅࡿࡇࡀ♧၀ࡉࢀࡓࠋ ௨ୖࡢࡇࡽ㸪W1DF ࡢᛶ㉁ࢆ᫂ࡽࡍࡿ┠ⓗ࡛ W2DF ༸⬊Ⓨ⫱ືែ㸪༸ ⮳ࡿࡲ࡛ࡢ༸⬊ቨ⾑ὶࡸ⾑୰ E2 ⃰ᗘ㸪LH ࢧ࣮ࢪ᭚㟢๓ᚋ࠾ࡅࡿ༸⬊ᾮᛶࢫࢸࣟࢻ ⃰ᗘ㸪㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿ LHr㸪ࢫࢸࣟࢻྜᡂ㓝⣲㸪⾑⟶సືᛶᅉᏊ࠾ࡼࡧ⾑ ⟶ᙧᡂᅉᏊࡢ mRNA Ⓨ⌧㔞㸪ࡉࡽࡣ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᙧែ࠾ࡼࡧᶵ⬟ࢆẚ㍑ ࡋࡓࡇࢁ㸪W1DF ࡣࢧࢬࡸᶵ⬟ᛶࡀ㧗ࡃ㸪άຊࡢ㧗࠸༸⬊࡛࠶ࡿࡇࡀ᫂ࡽ࡞ ࡗࡓࠋ 2㸫8㸬ᑠᣓ 㯤యᙧᡂᮇⓎ⫱ࡍࡿW1DFࡣ㸪㯤య㛤ⰼᮇⓎ⫱ࡍࡿW2DFẚ༸⬊ࡢⓎ⫱㸪༸⬊ ቨ⾑ὶ㸪༸๓༸⬊ࡢ㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿLHr㸪⾑⟶᪂⏕ᅉᏊ࠾ࡼࡧࢫࢸࣟࢻྜ ᡂ㓝⣲ࡢmRNAⓎ⌧㔞㸪ࢫࢸࣟࢻྜᡂ⬟㸪㯤యᙧᡂ࠾ࡼࡧ㯤యᶵ⬟ࡀ㧗ࡃ㸪άຊࡀ㧗࠸ ࡇࡀ♧၀ࡉࢀࡓࠋࡇࢀࡽࡢW1DFࡢᛶ㉁ࡣ㸪W1DFⓎ⫱ᮇ㛫୰ࡢ⾑୰P4⃰ᗘࡢప࠸᥎⛣ ࡼࡿࡶࡢ᥎ ࡉࢀࡓࠋ 32 W1DF Spontaneous ovulation Day0 B-mode US Day1 Day2 Day3 Day4 Color Doppler US Day5 Day6 W2DF Spontaneous ovulation Day7 Induced ovulation Day2 Day0 B-mode US Day1 Day2 Day3 Day4 Color Doppler US Day5 Day6 GnRH Fig. 2.1. Schematic diagram of experimental model of W1DF and W2DF in Experiment 1. In the first follicular wave, the day of spontaneous ovulation was defined as Day 0. In the second follicular wave, the day of spontaneous ovulation was defined as Day 7. Ovulation of the first-wave dominant follicle (DF) was induced by GnRH treatment on Day 2 to induce a new follicular wave, and induced ovulation was confirmed 36 h after GnRH treatment on Day 0. W1DF; DF in the first follicular wave. W2DF;DF in the second follicular wave. B-mode US = trans-rectal ultrasonography. Color Doppler US = trans-rectal color Doppler ultrasonography. 33 (B) (A) Fig. 2.2. Measuring method of blood flow area (BFA). Image of (A) is dominant follicle with blood flow signals in the follicular wall using color Doppler ultrasonography. The area of blood flow signals which is enclosed by white line is measured as BFA (image B). 34 Plasma P4Pconcentration of P4(ng/ml) Plasma (ng/ml) 4 concentration 8.0 7.0 W1DF W2DF 6.0 5.0 4.0 3.0 G䠖P < 0.001 D䠖P < 0.001 G,D䠖NS 2.0 1.0 0 Day0 Day1 Day2 Day3 Day4 Day5 Day6 Days from follicular wave emergence Fig. 2.3. Comparative changes of plasma P4 concentration of W1DF and W2DF from Day 0 to 6. Day 0 = The day of follicular wave emergence. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. Effect of group (G) (P < 0.001), day (D) (P < 0.001) and interaction between group and day (G,D) (NS). Values are mean r SEM of each point. 35 Dominant follicle diameter (mm) 17 16 15 14 13 W1DF W2DF G䠖P < 0.01 D䠖P < 0.01 G,D䠖P < 0.01 12 11 10 9 8 Day3 Day4 Day5 Day6 Days from follicular wave emergence Fig. 2.4. Comparative changes of dominant follicle (DF) of W1DF and W2DF from Day 3 to 6. Day 0 = The day of follicular wave emergence. W1DF; DF in the first follicular wave. W2DF; DF in the second follicular wave. Effect of group (G) (P < 0.01), day (D) (P < 0.01) and interaction between group and day (G,D) (P < 0.01). The asterisk denotes difference between each point, : P < 0.05. Values are mean r SEM of each point.㻌 36 Table 2.1. Blood flow in the follicular wall on Day6. W1DF W2DF P-value BFA(cm2) 0.32㼼0.05 0.21㼼0.05 BF% (%) 24.37㼼2.64 18.35㼼3.17 P < 0.1 P < 0.1 Values are mean 㼼 SEM. W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave. BFA = Blood flow area. BF% = The percentage of follicle circumference with blood flow signals. 37 W1DF Spontaneous ovulation Color Doppler US Day0 Day6 W2DF PGF2D Spontaneous ovulation Day7 Day7 Day8 Day9 GnRH Induced ovulation Day2 Day0 Color Doppler US Day6 GnRH PGF2D Day7 Day8 Day9 GnRH Fig. 2.5. Schematic diagram of experimental model of W1DF and W2DF in Experiment 2. In the first follicular wave, the day of spontaneous ovulation was defined as Day 0. To induce follicular maturation, PGF2α and GnRH were administrated on Day 6 and 8, respectively. In the second follicular wave, the day of spontaneous ovulation was defined as Day 7. Ovulation of the first-wave dominant follicle (DF) was induced by GnRH treatment on Day 2 to induce a new follicular wave, and induced ovulation was confirmed 36 h after GnRH treatment on Day 0. PGF2α and GnRH were administrated on Day 6 and 8, respectively. W1DF; DF in the first follicular wave. W2DF;DF in the second follicular wave. Color Doppler US = trans-rectal color Doppler ultrasonography. 38 Dominant follicle diameter (mm) 20 19 W1DF W2DF 18 17 16 15 G : P < 0.001 D : P < 0.001 G,D : NS 14 13 12 Day6 Day8 Day9 Days from follicular wave emergence Fig. 2.6. Comparative changes of dominant follicle (DF) of W1DF and W2DF from Day 6 to 9. Day 0 = The day of follicular wave emergence. W1DF; DF in the first follicular wave. W2DF;DF in the second follicular wave. Effect of group (G) (P < 0.001), day (D) (P < 0.001) and interaction between group and day (G,D) (NS). Values are mean r SEM of each point. 39 Plasma P4 P4 concentration concentration (ng/ml) (ng/ml) 8.0 W1DF W1DF 7.0 W2DF W2DF 6.0 5.0 G : P < 0.01 D : P < 0.01 G : P < 0.01 G,D : P < 0.05 D : P < 0.001 4.0 3.0 G,D : P < 0.05 䠆䠖P < 0.05 2.0 1.0 0 Day6 Day7 Day8 Day9 Days from follicular wave emergence Fig. 2.7. Comparative changes of plasma P4 concentration of W1DF and W2DF from Day 6 to 9. Day 0 = The day of follicular wave emergence. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave.㻌 Effect of group (G) (P < 0.01), day (D) (P < 0.01) and interaction between group and day (G,D) (P < 0.05). The asterisk denotes difference between each point, : P < 0.05. Values are mean r SEM of each point. 40 BFA 1.2 BFA(cm2) 1.0 0.8 G : P < 0.01 D : P < 0.01 G,D : P < 0.05 0.6 0.4 0.2 Day6 BF% Day8 70 Day9 W1DF W2DF BF% (%) 60 G : P < 0.01 D : P < 0.01 G,D : P < 0.01 50 40 30 20 10 Day6 Day8 Day9 Days from follicular wave emergence Fig. 2.8. Comparative changes of blood flow area (BFA) and the percentage of follicle circumference with blood flow signals (BF%) of W1DF and W2DF on Day 6 to 9. Cows were treated with PGF2α and GnRH on Day 6 and Day 8, respectively. Day 0 = day of follicular wave emergence. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. For BFA, effect of group (G) (P < 0.01), day (D) (P < 0.01) and interaction between group and day (G,D)(P < 0.05). For BF%, effect of group (G) (P < 0.01), day (D) (P < 0.01) and interaction between group and day (G,D) (P < 0.01). The asterisk denotes difference between each points, : P < 0.05, : P < 0.01. Values are mean r SEM of each point. 41 Table 2.2. Plasma E2 concentration on Day 8. E2 (pg/ml) W1DF W2DF P-value 7.5㼼0.9 4.4㼼0.5 P < 0.01 Values are mean 㼼 SEM. Day8 = The day of after 48h of PGF2α treatment. W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave. 42 Follicle aspilation W1DF Spontaneous ovulation Follicle aspilation Day0 Day6 Day7 Day8 Day9 Follicle aspilation W2DF PGF 2D Spontaneous ovulation Day 7 GnRH Follicle aspilation Induced ovulation Day 2 Day0 Day6 PGF2D GnRH Day7 Day8 Day9 GnRH Fig. 2.9. Schematic diagram of experimental model of W1DF and W2DF in Experiment 3. In the first follicular wave, the day of spontaneous ovulation was defined as Day 0. To induce follicular maturation, PGF2α and GnRH were administrated on Day 6 and 8, respectively. In the second follicular wave, the day of spontaneous ovulation was defined as Day 7. Ovulation of the first-wave dominant follicle (DF) was induced by GnRH treatment on Day 2 to induce a new follicular wave, and induced ovulation was confirmed 36 h after GnRH treatment on Day 0. PGF2α and GnRH were administrated on Day 6 and 8, respectively. W1DF; DF in the first follicular wave. W2DF;DF in the second follicular wave. DF were aspirated by transvaginal ultrasonography in different estrous cycle. 43 Table 2.3. Primers used in real-time PCR. Gene Sequence of nucleotide(5䇻-3䇻) Fa Rb Size(bp) Annealing Accesion No. temperature( q C) GCATCCACAAGCTTCCAGATGTTACGA GGGAAATCAGCGTTGTCCCATTGA 204 62 CYP19A1 F R TTGCAAAGCATCCCCAGGTT AGGTCCACAACGGGCTGGTA 151 60 Z32741 COX -2 F R TCCTGAAACCCACTCCCAACA TGGGCAGTCATCAGGCACAG 241 60 AF031698 PGFS F R GATCAAAGCGATTGCAGACA CAATGCGTTCAGGTGTCACT 113 59 S54973 PGES F R CGCTGCTGGTCATCAAAAT GGAAGGGGTAGATGGTCTCC 186 60 AY032727 iNOS F R TCATCTTCGCCACCAAGCAG CAGTGATGGCCGACCTGATG 180 60 U14640 eNOS F R GGAAATCGGGGGTCTGGAGT TTGGCGAGCTGAAAGCTGTG 219 60 M89952 VEGF120 F R CCCAGATGAGATTGAGTTCATTTT GCCTCGGCTTGTCACATTTT 377 56 M32976 VEGF164 F R CCCAGATGAGATTGAGTTCATTTT AGCAAGGCCCACAGGGATTT 245 58 M32976 F R GTGCATTTTGCCAATCACCT TTATTGAAAACGTGCCACCA 203 54.3 M174189 P450-scc F R CTGCAAATGGTCCCACTTCT CACCTGGTTGGGTCAAACTT 209 56.3 K02103 3β-HSD F R TCCACACCAGCACCATAGAA AAGGTGCCACCATTTTTCAG 118 56.3 X17614 F R CTCTCAAGGGCATTCTAGGC TGACAAAGTGGTCGTTGAGG 160 60 LHr StAR GAPDH a b F, forward R, reverse 44 NM 174381 NM001034034 Table 2.4. Sex steroids concentration in follicular fluid on Day 8. W1DF P4 (ng/ml) 37.4㼼8.5 E2 (ng/ml) 1,942.4㼼247.9 A4 (ng/ml) 161.8㼼26.2 W2DF P-value 34.6㼼2.2 NS 1,169.3㼼90.9 P < 0.05 104.6㼼12.7 P < 0.05 Values are mean 㼼 SEM. Day 8 = The day of after 40h of PGF2α treatment. W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave. 45 LHr CYP19A1 * 4.0 0.3 /GAPDH /GAPDH 3.0 0.2 0.1 0 2.0 1.0 0 W1DF W2DF W1DF W2DF Fig. 2.10. The relative amounts of mRNA of LHr and CYP19A1 in W1DF and W2DF granulosa cells on Day 8. In each figure, the black bar indicates W1DF and the white bar indicates W2DF. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. Day 8 = the day of after 40h of PGF2α treatment. The asterisk denotes differences, *: P < 0.05. Values are shown as mean r SEM. 㻌 46 Table 2.5. Follicular fluid sex steroids concentration on Day 9. W1DF W2DF P-value P4 (ng/ml) 90.1㼼11.8 91.4㼼7.7 NS E2 (ng/ml) 82.5㼼20.7 64.1㼼8.7 NS Values are mean 㼼 SEM. Day 9 = The day of after 24h of GnRH treatment. W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave. 47 COX-2 PGES 0.16 2.5 0.12 /GAPDH /GAPDH 2.0 1.5 1.0 0.08 0.5 0.04 0 0 W1DF W1 W2DF W2 W1 W2DF W2 W1DF PGFS /GAPDH 0.8 0.6 0.4 0.2 0 W1DF W1 W2DF W2 Fig. 2.11. The relative amounts of mRNA of COX-2, PGES and PGFS of W1DF and W2DF granulosa cells on Day 9. In each figure, the black bar indicates W1DF and the white bar indicates W2DF. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. Day9 = the day of after 24h of GnRH treatment. Values are shown as mean r SEM. 㻌 48 iNOS eNOS ͤ . 0.012 0.02 /GAPDH /GAPDH 0.025 0.015 0.01 0.08 0.04 0.005 0 0 W1DF W1 W1DF W1 W2DF W2 W2DF W2 Fig. 2.12. The relative amounts of mRNA of iNOS and eNOS in W1DF and W2DF granulosa cells on Day 9. In each figure, the black bar indicates W1DF and the white bar indicates W2DF. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. Day9 = the day of after 24h of GnRH treatment. The asterisk denotes differences, shown as mean r SEM. 㻌 49 䈜: P < 0.1. Values are VEGF120 VEGF164 * ͤ 1.4 0.5 1.2 0.4 /GAPDH /GAPDH 1.0 0.8 0.6 0.3 0.2 0.4 0.1 0.2 0 0 W1 W1DF W1 W1DF W2 W2DF W2 W2DF FGF-2 ͤ 2.5 /GAPDH 2.0 1.5 1.0 0.5 0 W1 W1DF W2 W2DF Fig. 2.13. The relative amounts of mRNA of VEGF120, VEGF164 and FGF-2 in W1DF and W2DF granulosa cells on Day 9. In each figure, the black bar indicates W1DF and the white bar indicates W2DF. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. Day9 = the day of after 24h of GnRH treatment. The asterisk denotes differences, 䈜: shown as mean r SEM. 㻌 50 P < 0.1, *: P < 0.05. Values are StAR P450-scc ͤ 16 25 14 20 12 /GAPDH /GAPDH * 18 30 15 10 8 6 10 4 5 2 0 0 W1 W1DF W2 W2DF W1 W1DF W2 W2DF 3β-HSD * 140 120 /GAPDH 100 80 60 40 20 0 W1 W1DF W2 W2DF Fig. 2.14. The relative amounts of mRNA of StAR, P450-scc, and 3E-HSD in W1DF and W2DF granulosa cells on Day 9. In each figure, the black bar indicates W1DF and the white bar indicates W2DF. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. Day9 = the day of after 24h of GnRH treatment. The asterisk denotes differences, 䈜: P < 0.1, *: P < 0.05. Values are shown as mean r SEM. 㻌 51 Ovulation Color Doppler US W1CL W2CL Day10 Day11 Day12 Day13 Day14 Day15 Day16 Day17 Day18 Day19 Day20 Day21 Fig. 2.15. Schematic diagram of experimental model of W1CL and W2CL in Experiment 4. Day 0 = The day of spontaneous ovulation. Day10 = The day of W1DF or W2DF ovulation. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. W1CL; Corpus luteum (CL) formed after ovulation of W1DF. W2CL; CL formed after ovulation of W2DF. B-mode US = transrectal ultrasonography. Color Doppler US = trans-rectal color Doppler ultrasonography. 52 Table 2.6. Diameter and blood flow in the follicular wall on Day 9. W1DF W2DF P-value Diameter (mm) 17.7㼼0.3 15.7㼼0.5 P 0.01 BF%(%) 55.3㼼4.0 36.1㼼2.3 P 0.01 Values are mean 㼼 SEM. Day 9 = The day of after 24h of GnRH treatment. W1DF = Dominant follicle (DF) in the first follicular wave. W2DF = DF in the second follicular wave. BF% = The percentage of follicle circumference with blood flow signals. 53 Cross-sectional area of corpus luteum (mm2) 䠆 700 W1CL W2CL 600 䠆 䠆 500 400 300 G : P < 0.01 D : P < 0.01 200 G,D : P < 0.05 100 0 12 15 18 21 Days from follicular wave emergence Fig. 2.16. Comparative changes of cross-sectional area of corpus luteum (CL) of W1CL and W2CL on Day 12, 15, 18 and 21. Day 10 = ovulation day of W1DF and W2DF. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. W1CL; CL formed after ovulation of W1DF. W2CL; CL formed after ovulation of W2DF. Effect of group (G) (P < 0.01), day (D) (P < 0.01) and interaction between group and day (G,D)(P < 0.05). The asterisk denotes difference between each point, : P < 0.05. Values are mean r SEM of each point.㻌 54 Blood flow area of corpus luteum (mm2) 500 W1CL W2CL 400 䠆 䠆 300 G : P < 0.05 D : P < 0.05 G,D : P < 0.05 200 100 0 12 15 18 21 Days from follicular wave emergence Fig. 2.17. Comparative changes of blood flow area in the corpus luteum (CL) of W1CL and W2CL on Day 12, 15, 18 and 21. Day 10 = ovulation day of W1DF and W2DF. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. W1CL; CL formed after ovulation of W1DF. W2CL; CL formed after ovulation of W2DF. Effect of group (G) (P < 0.05), day (D) (P < 0.05) and interaction between group and day (G,D)(P < 0.05). The asterisk denotes difference between each point, : P < 0.05. Values are mean r SEM of each point. 㻌 55 Plasma P4 concentration (ng/ml) 7.0 W1CL W2CL 6.0 5.0 4.0 3.0 G : P < 0.05 G:P<0.05 D:P<0.05 G,D : NS GD:NS 2.0 D : P < 0.05 1.0 0 10 11 12 13 14 15 16 17 18 19 20 21 Days from follicular wave emergence Fig. 2.18. Comparative changes in the plasma P4 concentration of W1CL and W2CL. Blood samples were collected daily from Day 10 to 21. Day 10 = ovulation day of W1DF and W2DF. W1DF; Dominant follicle (DF) in the first follicular wave. W2DF;DF in the second follicular wave. W1CL; Corpus luteum (CL) formed after ovulation of W1DF. W2CL; CL formed after ovulation of W2DF. Effect of group (G) (P < 0.05), day (D) (P < 0.05) and interaction between group and day (G,D)(NS). Values are mean r SEM of each point. 㻌 56 ➨㸱❶ ➨ 1 ༸⬊Ἴᖍ༸⬊㸦W1DF㸧ࡢⓎ⫱ࢆไᚚࡍࡿせᅉ 57 3㸫1㸬ᗎㄽ ➨ 2 ❶ࡼࡾ㸪W1DF ࡢⓎ⫱ືែ㸪ᶵ⬟㸪༸๓ᚋࡢᛶ㉁࠾ࡼࡧ༸ᚋᙧᡂࡉࢀࡿ㯤 యࡢᛶ㉁ࢆ W2DF ẚ㍑ࡍࡿࡇ࡛᫂ࡽࡋࡓࠋࡲࡓ㸪୧⪅ࡢᛶ㉁ࡢ㐪࠸ࢆࡶࡓࡽࡍ せᅉࡋ࡚㸪ᮎᲈ⾑୰ࡢ P4 ⃰ᗘࡢ㐪࠸ࡼࡿ LH ࣃࣝࢫ㢖ᗘࡢ㐪࠸ࡀ᥎ ࡉࢀࡓ㸦15㸪 52㸧 ࠋAdams ࡽ㸦5㸧ࡣⓎ┤ᚋࡢ࢘ࢩ⭐ෆ␃⨨ᆺ P4 〇ࢆᤄධࡋࡓࡁ㸪W1DF ࡢ ᭱┤ᚄࡀ↓ฎ⨨⩌ẚ㍑ࡋ࡚ᑠࡉ࠸ࡇࢆ♧ࡋ࡚࠸ࡿࠋࡇࡢࡇࡽ㸪W1DF ࡢᛶ㉁ ࢆᕥྑࡍࡿせᅉࡋ࡚㸪Ⓨ⫱ᮇ㛫୰ࡢ⾑୰ P4 ⃰ᗘࡼࡿᙳ㡪ࡀ⪃࠼ࡽࢀࡿࠋࡋࡋ࡞ࡀ ࡽ㸪↓ฎ⨨≧ែ࡛ࡢ W1DF ࡢⓎ⫱ࢆไᚚࡍࡿせᅉࡣ㸪㐣ཤ࠾࠸࡚༑ศ᫂ࡽࡉࢀ ࡚ࡣ࠸࡞࠸ࠋࡑࡇ࡛㸪ᮏ❶࡛ࡣ༸⬊Ⓨ⫱ᮇ㛫୰ࡢ⾑୰ P4 ⃰ᗘࡀ༸⬊ࡢⓎ⫱ཬࡰࡍᙳ㡪 ࢆㄪࡓࠋࡉࡽ㸪W1DF ࡣ㯤యྠ୍༸ᕢෆ࡛Ⓨ⫱ࡍࡿ㸦ඹᏑ㸧㸪ᑐഃࡢ༸ᕢ࡛ Ⓨ⫱ࡍࡿ㸦㠀ඹᏑ㸧ࡢࡕࡽ࡛࠶ࡿࠋࡇࢀࡲ࡛㸪W1DF 㯤యࡢඹᏑࡲࡓࡣ㠀ඹᏑ ࡢⓎ⏕⋡ࡣኚࢃࡽ࡞࠸࠸࠺ሗ࿌ࡀ࠶ࡾ㸦40㸧 㸪୍᪉࡛ඹᏑࡢⓎ⏕⋡ࡀ㧗࠸࠸࠺ሗ࿌ࡶ ࠶ࡿ㸦33㸧 ࠋࡋࡋ࡞ࡀࡽ㸪W1DF 㯤యࡢඹᏑࡲࡓࡣ㠀ඹᏑࡀ W1DF ࡢⓎ⫱ືែࡸ༸ ⬊ࡢᶵ⬟࠼ࡿᙳ㡪ࡣ༑ศࡣ᳨ウࡉࢀ࡚࠸࡞࠸ࠋࡑࡇ࡛㸪ᮏ❶࡛ࡣ㸪༸ᕢෆᒁᡤ⎔ ቃࡀ༸⬊ࡢᛶ㉁ᙳ㡪ࢆ࠼ࡿྍ⬟ᛶࢆ᥈ࡿࡓࡵ㸪ඹᏑ㠀ඹᏑࡢ㛫࡛ࡢ W1DF ࡢⓎ ⫱ືែ㸪༸⬊ᾮ୰ࢫࢸࣟࢻ࣍ࣝࣔࣥࢆ ᐃࡋ㸪ẚ㍑ࡋࡓࠋ 3㸫2㸬ヨ㦂 1㸸༸⬊ἼⓎ⌧ࡽ 7 ᪥┠ࡢ W1DF ࡢ┤ᚄ⾑୰ P4 ⃰ᗘࡢ┦㛵㛵ಀ࠾ࡼࡧ W1DF 㯤యࡢඹᏑࡲࡓࡣ㠀ඹᏑ㛵ಀࡀ W1DF ࡢ┤ᚄཬࡰࡍᙳ㡪 3㸫2㸫1㸬౪ヨື≀ 58 ⮫ᗋⓗᗣ࡛㸪ṇᖖ࡞Ⓨ࿘ᮇࢆ᭷ࡍࡿ࣍ࣝࢫࢱࣥ✀㠀Ἢங∵ 30 㢌㸦ᖹᆒᖺ㱋㸸 6.1 r 0.4 ṓ㸪ᖹᆒ±ᶆ‽ㄗᕪ) ࢆ⏝࠸࡚㸪ᘏ 44 㢌ࡢࢹ࣮ࢱࢆゎᯒ⏝ࡋࡓࠋ࢘ࢩࡣ ᖏᗈ␆⏘Ꮫ␆⏘ࣇ࣮ࣝࢻ⛉Ꮫࢭࣥࢱ࣮ෆࡢࣃࢻࢵࢡ࡚㸪ࢥ࣮ࣥࢧ࣮ࣞࢪ㸪ⲡ ࠾ࡼࡧ⃰ཌ㣫ᩱࡀΰྜࡉࢀࡓ TMR ࢆ⤥ࡉࢀ㸪 ⮬⏤㣧Ỉ࡛ࡁࡿ≧ἣ࡛㣫㣴ࡉࢀ࡚࠸ࡓࠋ ᐇ㦂ෆᐜࡣᖏᗈ␆⏘Ꮫࡢᐇ㦂⌮ጤဨ࡚ᢎㄆࡉࢀ࡚࠸ࡿࠋ 3㸫2㸫2㸬ᐇ㦂᪉ἲ ⮬↛Ⓨ㛤ጞᚋ㸪12 㛫ࡈ㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚༸ᕢࢆほᐹࡋ㸪༸๓༸ ⬊ࡢᾘኻࢆ☜ㄆࡋࡓⅬࢆ Day 0 ࡋࡓࠋDay 6 ㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚ W1DF ࡢほᐹࢆ⾜ࡗࡓࠋࡇࡢ㸪㯤య W1DF ࡀྠ୍༸ᕢෆᏑᅾࡍࡿඹᏑ㛵ಀ㸪ࡑࢀࡒࢀ ࡀูࡢ༸ᕢᏑᅾࡍࡿ㠀ඹᏑ㛵ಀ࡛࠶ࡿࡢࢆ☜ㄆࡋࡓࠋࡲࡓ㸪29 㢌ࡽṇ୰ᑿ㟼⬦ࡼ ࡾ᥇⾑ࢆ⾜ࡗࡓࠋ 3㸫2㸫3㸬 ༸⬊ᙧែࡢほᐹ ᖍ༸⬊ࡢᖹᆒ┤ᚄࡣ㸪5.0MHzࡢࣜࢽࣉ࣮ࣟࣈࡀᒓࡋࡓ⤒┤⭠㉸㡢Ἴ⏬ീデ᩿ ⨨㸦HS̺101V㸪ᮏከ㟁Ꮚ㸪ឡ▱㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋࣉ࣮ࣟࣈࢆᡭᣦࡶ┤⭠ෆ ᤄධࡋ㸪ᕥྑ୧ഃ༸ᕢࡢ᩿ᒙീࢆほᐹࡋࡓࠋᖍ༸⬊ࡢ༸⬊᩿ᒙ⏬ീࡀ᭱࡞ࡿࡼ࠺㟼 Ṇ⏬ࢆグ㘓ࡋ㸪ࣃࢯࢥࣥୖ㟼Ṇ⏬ࢆ⛣ືࡉࡏ㸪ᖍ༸⬊ࡣ㛗ᚄ▷ᚄࡢᖹᆒ್ࢆᖹᆒ┤ ᚄࡋ㸪ࢹ࣮ࢱࡋ࡚⏝࠸ࡓࠋ 3㸫2㸫4㸬᥇⾑ 21G᥇⾑㔪࣊ࣃࣜࣥຍ┿✵᥇⾑⟶㸦࣋ࣀࢪ࢙ࢡࢺϩ┿✵᥇⾑⟶㸪TERUMO㸪ᮾி㸪᪥ ᮏ㸧ࢆ⏝࠸࡚ᑿ㟼⬦ࡼࡾ10mlࡢ᥇⾑ࢆ⾜ࡗࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣịỈ୰࡚ᛴ෭ᚋ㸪4Υ࡛ 2,000 g㸪20ศ㛫㐲ᚰࡋ㸪⾑₢ศ㞳ᚋ㸪⾑୰P4⃰ᗘ ᐃࡲ࡛㸫30Υ࡚⤖ಖᏑࡋࡓࠋ 59 3㸫2㸫5㸬⾑୰P4⃰ᗘࡢ ᐃ Day 6࠾ࡅࡿ⾑୰P4⃰ᗘࡣEIAࡼࡾ ᐃࡋࡓࠋ ᐃࡣ➨2❶ྠᵝࡢ᪉ἲ࡛⾜ࡗࡓࠋ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀࡒࢀ5.2㸣࠾ࡼࡧ7.4%࡛࠶ࡗࡓࠋ 3㸫2㸫6㸬⤫ィฎ⌮ Day 6 ࠾ࡅࡿᖍ༸⬊ࡢᖹᆒ┤ᚄ⾑୰ P4 ⃰ᗘࡢ┦㛵ᛶࢆㄪࡿࡓࡵࡣ㸪᭱ᑠ 2 ἲࡼࡾᅇᖐ┤⥺ᘧࢆồࡵ㸪Pearson ࡢ┦㛵ಀᩘࢆồࡵ࡚┦㛵ࡢ᭷ពᛶࢆ᳨ᐃࡋࡓࠋ⤖ ᯝࡣࡍ࡚ᖹᆒ್±ᶆ‽ㄗᕪ࡛♧ࡋࡓࠋඹᏑ⩌㠀ඹᏑ⩌ࡢ㛫࡛ Day 6 ࠾ࡅࡿᖍ༸⬊ ࡢᖹᆒ┤ᚄ⾑୰ P4 ⃰ᗘࡢẚ㍑ࡣ㸪F ᳨ᐃࢆ⾜࠸ศᩓࡢᆒ୍ᛶࢆ☜ㄆࡋࡓᚋ㸪Student ࡢ t ᳨ᐃࢆ⏝࠸ࡓࠋP < 0.05 ࡢሙྜࢆ᭷ពᕪ࠶ࡾࡋࡓࠋ⤫ィฎ⌮ࡣ⤫ィฎ⌮ࢯࣇࢺ࡛ ࠶ࡿ StatView㸦Abacus Concepts㸪Inc.㸪Berkeley㸪CA㸪USA㸧ࢆ⏝࠸ࡓࠋ 3㸫3㸬ヨ㦂 2㸸W1DF 㯤యࡢඹᏑࡲࡓࡣ㠀ඹᏑ㛵ಀࡀ༸⬊ἼⓎ⌧᪥ࡽ 5 ᪥┠ࡲ࡛ࡢ W1DF 㯤యࡢⓎ⫱ືែ㸪⾑୰ P4 ⃰ᗘ㸪࠾ࡼࡧ༸⬊ᾮࢫࢸࣟࢻ࣍ࣝࣔࣥ⃰ᗘ ཬࡰࡍᙳ㡪 3㸫3㸫1㸬౪ヨື≀ ⮫ᗋⓗᗣ࡛㸪ṇᖖ࡞Ⓨ࿘ᮇࢆ᭷ࡍࡿ࣍ࣝࢫࢱࣥ✀㠀Ἢங∵ 5 㢌㸦ᖹᆒᖺ㱋㸸 5.6 r 1.0 ṓ㸪ᖹᆒ±ᶆ‽ㄗᕪ) ࢆ⏝࠸ࡓࠋ࢘ࢩࡣᖏᗈ␆⏘Ꮫ␆⏘ࣇ࣮ࣝࢻ⛉Ꮫࢭࣥࢱ ࣮ෆࡢࣃࢻࢵࢡ࡚㸪ࢥ࣮ࣥࢧ࣮ࣞࢪ㸪ⲡ࠾ࡼࡧ⃰ཌ㣫ᩱࡀΰྜࡉࢀࡓ TMR ࢆ⤥ ࡉࢀ㸪⮬⏤㣧Ỉ࡛ࡁࡿ≧ἣ࡛㣫㣴ࡉࢀ࡚࠸ࡓࠋᐇ㦂ෆᐜࡣᖏᗈ␆⏘Ꮫࡢᐇ㦂⌮ጤ ဨ࡚ᢎㄆࡉࢀ࡚࠸ࡿࠋ 60 3㸫3㸫2㸬ᐇ㦂᪉ἲ ⮬↛Ⓨ㛤ጞᚋ㸪12 㛫ࡈ㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚༸ᕢࢆほᐹࡋ㸪༸๓༸ ⬊ࡢᾘኻࢆ☜ㄆࡋࡓⅬࢆ Day 0 ࡋࡓࠋ Day 0 ࡽ 4 ࡅ࡚㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝ ࠸࡚ W1DF ࠾ࡼࡧ㯤యࡢほᐹࢆ⾜ࡗࡓࠋࡲࡓ㸪Day 4 ࡢⅬ࡛ W1DF ࡀ㯤యྠ୍༸ᕢ ෆᏑᅾࡍࡿಶయࢆඹᏑ⩌㸪W1DF ࡀ㯤యᑐഃࡢ༸ᕢᏑᅾࡍࡿሙྜࢆ㠀ඹᏑ⩌ ศ㢮ࡋࡓࠋࡲࡓ㸪Day 0 ࡽ 4 ࡅ࡚㐃᪥ṇ୰ᑿ㟼⬦ࡼࡾ᥇⾑ࢆ⾜ࡗࡓࠋࡑࡋ࡚㸪Day 4 W1DF ࡢ༸⬊ᾮ࠾ࡼࡧ㢛⢏ᒙ⣽⬊ࡢᅇࢆ⤒⭐༸⬊྾ᘬἲࡼࡾ⾜ࡗࡓࠋ 5 㢌ࡢ౪ヨ∵ࡣ㸪ࣛࢸࣥ᱁Ꮚἲࡼࡿࢡࣟࢫ࣮࢜ࣂ࣮ヨ㦂ࢆ⾜ࡗࡓࡓࡵ㸪ඹᏑ⩌㸦n㸻 5㸧࠾ࡼࡧ㠀ඹᏑ⩌㸦n㸻5㸧ࡣࡑࢀࡒࢀ 5 㢌ࡢ౪ヨ∵ࡀ 1 ᅇࡎࡘྵࡲࢀ࡚࠸ࡿࠋ 3㸫3㸫3㸬༸⬊࠾ࡼࡧ㯤యᙧែࡢほᐹ ᖍ༸⬊ࡢᖹᆒ┤ᚄ࠾ࡼࡧ㯤యࡢ᩿㠃✚ࡣ㸪5.0MHzࡢࣜࢽࣉ࣮ࣟࣈࡀᒓࡋࡓ⤒┤ ⭠㉸㡢Ἴ⏬ീデ᩿⨨㸦HS̺101V㸪ᮏከ㟁Ꮚ㸪ឡ▱㸪᪥ᮏ㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋࣉ࣮ࣟ ࣈࢆᡭᣦࡶ┤⭠ෆᤄධࡋ㸪ᕥྑ୧ഃ༸ᕢࡢ᩿ᒙീࢆほᐹࡋࡓࠋᖍ༸⬊ࡢ༸⬊᩿ ᒙ⏬ീࡀ᭱࡞ࡿࡼ࠺㟼Ṇ⏬ࢆグ㘓ࡋ㸪ࣃࢯࢥࣥୖ㟼Ṇ⏬ࢆ⛣ືࡉࡏ㸪ᖍ༸⬊ࡣ㛗 ᚄ▷ᚄࡢᖹᆒ್ࢆᖹᆒ┤ᚄࡋ㸪ࢹ࣮ࢱࡋ࡚⏝࠸ࡓࠋ㯤యࡢ᩿㠃✚ࡣ㸪㯤యࡢ᩿ᒙ⏬ ീࡀ᭱࡞ࡿࡼ࠺㟼Ṇ⏬ࢆグ㘓ࡋࡓࠋࣃࢯࢥࣥୖ㟼Ṇ⏬ࢆ⛣ືࡉࡏ㸪㯤యࡢ㛗ᚄ▷ ᚄࡢᖹᆒ್ࡽᖹᆒ┤ᚄࢆ⟬ฟࡋ㸪 㸦ᖹᆒ┤ᚄ㸭2㸧2×3.14ᙜ࡚ࡣࡵ㯤య᩿㠃✚ࢆ⟬ฟࡋ㸪 ࢹ࣮ࢱ⏝࠸ࡓࠋ 3㸫3㸫4㸬᥇⾑ 21G᥇⾑㔪࣊ࣃࣜࣥຍ┿✵᥇⾑⟶㸦࣋ࣀࢪ࢙ࢡࢺϩ┿✵᥇⾑⟶㸪TERUMO㸪ᮾி㸪᪥ ᮏ㸧ࢆ⏝࠸࡚ᑿ㟼⬦ࡼࡾ10mlࡢ᥇⾑ࢆ⾜ࡗࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣịỈ୰࡚ᛴ෭ᚋ㸪4Υ࡛ 61 2,000 g㸪20ศ㛫㐲ᚰࡋ㸪⾑₢ศ㞳ᚋ㸪⾑୰P4࠾ࡼࡧE2⃰ᗘ ᐃࡲ࡛㸫30Υ࡚⤖ಖᏑࡋ ࡓࠋ 3㸫3㸫5㸬ᖍ༸⬊ࡢ྾ᘬ༸⬊ᾮࡢᅇ ᖍ༸⬊ࡢ྾ᘬ㸪༸⬊ᾮࡢᅇ࠾ࡼࡧࡑࡢᚋࡢฎ⌮ಖᏑࡣ㸪➨2❶ྠᵝࡢ᪉ἲ࡛⾜ ࡗࡓࠋ 3㸫3㸫6㸬༸⬊ᾮP4㸪E2࠾ࡼࡧA4⃰ᗘࡢ ᐃ ༸⬊ᾮࢫࢸࣟࢻ࣍ࣝࣔࣥࡢ ᐃࡣ➨2❶ྠᵝࡢ᪉ἲ࡛⾜ࡗࡓࠋࡇࡢ ᐃἲࡼࡿP4㸪 E2࠾ࡼࡧA4ࡢᢳฟຠ⋡ࡣࡑࢀࡒࢀ85㸣㸪81㸣࠾ࡼࡧ84㸣࡛࠶ࡗࡓࠋP4ࡢ ᐃෆኚື࠾ࡼࡧ ᐃෆኚືࡣࡑࢀࡒࢀ5.5%࠾ࡼࡧ7.4%㸪E2ࡢ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣ6.2㸣࠾ࡼࡧ 8.6%㸪A4ࡢ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀࡒࢀ5.6㸣࠾ࡼࡧ7.8%࡛࠶ࡗࡓࠋ 3㸫3㸫7㸬⾑୰P4࠾ࡼࡧE2⃰ᗘࡢ ᐃ Day 0ࡽ4࠾ࡅࡿ⾑୰P4⃰ᗘ࠾ࡼࡧDay 4ࡢ⾑୰E2⃰ᗘࡣEIAࡼࡾ ᐃࡋࡓࠋ ᐃ᪉ ἲࡣ➨2❶ྠᵝࡢ᪉ἲ࡛⾜ࡗࡓࠋP4ࡢ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀࡒࢀ5.2㸣࠾ࡼ ࡧ7.4%㸪E2ࡢ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀࡒࢀ6.0㸣࠾ࡼࡧ8.4㸣࡛࠶ࡗࡓࠋ 3㸫3㸫8㸬⤫ィฎ⌮ ⤖ᯝࡣࡍ࡚ᖹᆒ್±ᶆ‽ㄗᕪ࡛♧ࡋࡓࠋ⾑୰ P4 ⃰ᗘ㸪ᖍ༸⬊ࡢᖹᆒ┤ᚄ࠾ࡼࡧ㯤 య᩿㠃✚ࡢ᥎⛣ࡢẚ㍑ࡣ㸪2 ඖ㓄⨨ศᩓศᯒࢆ⾜࠸యࡢኚࡢ᭷ពᛶࢆㄪ㸪 స⏝ࡀ᭷ព࡛࠶ࡗࡓࡶࡢᑐࡋ࡚㸪Bartlet ᳨ᐃࢆ⾜࠸ศᩓࡢᆒ୍ᛶࢆ᳨ᐃࡋࡓᚋ㸪 Post-hoc ᳨ᐃࡋ࡚ Schefe ࡢከ⩌ẚ㍑᳨ᐃࢆ⾜ࡗࡓࠋDay 4 ࠾ࡅࡿඹᏑ⩌㠀ඹᏑ⩌ 㛫࡛ࡢ⾑୰ E2 ⃰ᗘ࠾ࡼࡧ༸⬊ᾮ୰ P4㸪E2 ࠾ࡼࡧ A4 ⃰ᗘࡢ᳨ᐃࡣ F ᳨ᐃࡼࡾศᩓࡢ 62 ᆒ୍ᛶࢆ᳨ᐃࡋ㸪Student ࡢ t ᳨ᐃࢆ⾜ࡗࡓࠋP < 0.05 ࡢሙྜࢆ᭷ពᕪ࠶ࡾࡋࡓࠋ⤫ィ ฎ⌮ࡣ⤫ィฎ⌮ࢯࣇࢺ࡛࠶ࡿ StatView㸦Abacus Concepts㸪Inc.㸪Berkeley㸪CA㸪USA㸧 ࢆ⏝࠸ࡓࠋ 3㸫4㸬⤖ᯝ 3㸫4㸫1㸬ヨ㦂 1 ⤖ᯝ ᮏヨ㦂࡛ඹᏑ⩌ࡣ n㸻22㸦ᖹᆒᖺ㱋㸸6.2±0.6㸧 㸪㠀ඹᏑ⩌ࡣ n㸻22㸦ᖹᆒᖺ㱋㸸6.2±0.4㸧 ࡛࠶ࡗࡓࠋ᥇⾑ࢆ⾜ࡗࡓ࢘ࢩ㸦n㸻29㸪ᖹᆒᖺ㱋㸸6.1±0.4㸧ࡢ⾑୰ P4 ⃰ᗘࡢᖹᆒ್ࡣ 3.0±0.2 ng/ml ࡛࠶ࡗࡓࠋࡲࡓ㸪ඹᏑ⩌㸦n㸻16㸪ᖹᆒᖺ㱋㸸6.0±0.7㸧࡛ࡢ⾑୰ P4 ⃰ᗘࡢᖹᆒ್ࡣ 㠀ඹᏑ⩌㸦n㸻13㸪ᖹᆒᖺ㱋㸸6.2±0.6㸧࡛ࡢ⾑୰ P4 ⃰ᗘࡢᖹᆒ್ẚ࡚㸪᭷ពప ࡗࡓ㸦Table 3.1.㸧 ࠋ Day 6 ࠾ࡅࡿ⾑୰ P4 ⃰ᗘᖍ༸⬊┤ᚄࡢ┦㛵㛵ಀࢆㄪࡓ⤖ᯝ㸪᭷ព࡞┦㛵㛵 ಀࡣほᐹࡉࢀ࡞ࡗࡓ㸦Fig. 3.1.㸧 ࠋࡲࡓ㸪ඹᏑ⩌㠀ඹᏑ⩌ࢆࡑࢀࡒࢀศࡅ࡚┦㛵㛵ಀ ࢆㄪࡓሙྜ㸪୧⩌ࡶ⾑୰ P4 ⃰ᗘᖍ༸⬊┤ᚄࡢ㛫᭷ព࡞┦㛵㛵ಀࡣほᐹࡉࢀ ࡞ࡗࡓ㸦Fig. 3.2.㸧 ࠋ ඹᏑ⩌㠀ඹᏑ⩌࡛ Day 6 ࠾ࡅࡿᖍ༸⬊ࡢᖹᆒ┤ᚄࢆẚ㍑ࡋࡓࡇࢁඹᏑ⩌࡛᭷ ពࡁࡗࡓ㸦Table 3.2.㸧 ࠋ 3㸫4㸫2㸬ヨ㦂 2 ⤖ᯝ ඹᏑ⩌㠀ඹᏑ⩌ࡢᖍ༸⬊ࡢᖹᆒ┤ᚄࡢ᥎⛣ࢆ Fig. 3.3.♧ࡋࡓࠋඹᏑ⩌࡛ Day 2 ࡽ 4 ࡅ࡚㠀ඹᏑ⩌ẚ㍑ࡋ࡚ࡁࡃ᥎⛣ࡋ࡚࠸ࡓࡀ㸪᭷ព࡞ᕪࡣࡳࡽࢀ࡞ࡗࡓࠋ ඹᏑ⩌㠀ඹᏑ⩌ࡢ㯤య᩿㠃✚࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ࢆ Fig. 3.4. Fig. 3.5.ࡑࢀࡒࢀ♧ ࡋࡓࠋ㯤య᩿㠃✚࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ඹ୧⩌㛫࡛ᕪࡣ࡞ࡗࡓࠋ 63 Day 4 ࠾ࡅࡿ⾑୰ E2 ⃰ᗘࢆẚ㍑ࡋࡓࡇࢁ㸪ඹᏑ⩌࡛㧗࠸ഴྥࢆ♧ࡋࡓࠋDay 4 ࠾ࡅࡿ༸⬊ᾮ୰ࡢᛶࢫࢸࣟࢻ࣍ࣝࣔࣥ⃰ᗘࢆ Table 3.4.♧ࡋࡓࠋ༸⬊ᾮ E2 ⃰ᗘ࠾ࡼࡧ A4 ⃰ᗘᕪࡣࡳࡽࢀ࡞ࡗࡓࡀ㸪༸⬊ᾮ P4 ⃰ᗘࡣඹᏑ⩌࡛᭷ព㧗ࡗࡓࠋ 3㸫5㸬⪃ᐹ ᮏヨ㦂࠾࠸࡚㸪ヨ㦂 1 ࡽ Day 6 ࡢ⾑୰ P4 ⃰ᗘࡣ W1DF ࡢ┤ᚄᙳ㡪ࢆ࠼࡞࠸ࡇ ࢆ♧ࡋࡓࡀ㸪W1DF 㯤యࡢ༸ᕢෆ࡛ࡢඹᏑࡲࡓࡣ㠀ඹᏑ㛵ಀࡀᖍ༸⬊ࡢ┤ᚄࡸ ⾑୰ P4 ⃰ᗘᙳ㡪ࢆ࠼ࡿࡇࡀ♧ࡉࢀࡓࠋࡉࡽ㸪ヨ㦂 2 ࡛ࡣඹᏑ⩌࠾࠸࡚㸪⾑୰ E2 ⃰ᗘ࠾ࡼࡧ༸⬊ᾮ୰ P4 ⃰ᗘࡀ㧗್ࢆ♧ࡋࡓࡇࡽ㸪W1DF ࡢⓎ⫱ࡸᛶ㉁ࢆไᚚࡍࡿ せᅉࡋ࡚㸪༸ᕢෆ࡛ࡢ㯤యࡢඹᏑࡲࡓࡣ㠀ඹᏑ≧ែࡀࡁࡃᙳ㡪ࢆཬࡰࡍࡇࡀ᫂ ࡽ࡞ࡗࡓࠋ ➨ 2 ❶࠾࠸࡚㸪 W1DF W2DF ࡢẚ㍑࡛ࡣᖍ༸⬊ࡢⓎ⫱ࡣ⾑୰ P4 ⃰ᗘࡀࡁࡃ 㛵ࡍࡿࡇࢆ♧ࡋࡓࡀ㸪Day 6 ࠾ࡅࡿ W1DF ࡢᖹᆒ┤ᚄ⾑୰ P4 ⃰ᗘ┦㛵㛵ಀࡣ ࡳࡽࢀ࡞ࡗࡓࠋ⮬↛Ⓨᚋࡢ༸☜ㄆ⭐ෆ␃⨨ᆺ P4 〇ࢆᤄධࡋࡓࡁࡢ㸪༸ᕢ ෆ࡛Ⓨ⫱ࡍࡿ W1DF ࡢⓎ⫱ືែࢆほᐹࡋࡓ◊✲࡛ࡣ㸪ᤄධࡋ࡞ࡗࡓ∵ẚ᭱㐩 ࡋࡓࡁࡢᖹᆒ┤ᚄࡀᑠࡉ࠸ࡇࢆሗ࿌ࡋ࡚࠸ࡿ㸦5㸧 ࠋࡇࡢࡇࡣ㸪ᖍ༸⬊Ⓨ⫱ᮇ㛫 ୰ࡢ⾑୰ P4 ⃰ᗘࡼࡾᖍ༸⬊ࡢⓎ⫱ࡀไᚚࡉࢀࡿࡇࢆពࡋ࡚࠸ࡿࠋࡋࡋ࡞ࡀࡽ㸪 ୖグࡢሗ࿌ࡣ㸪⭐ෆ␃⨨ᆺ P4 〇ࢆᤄධࡋࡓ࢘ࢩࢆ⏝࠸ࡓヨ㦂࡛࠶ࡾ㸪W1DF ࡢⓎ⫱ᮇ 㛫୰ࢆ㏻ࡋ࡚ᖖ⾑୰ P4 ⃰ᗘࡣ㧗್࡛࠶ࡿࡇࡽ㸪 ➨ 2 ❶࠾ࡅࡿ W2DF ࡢ㧗࠸⾑୰ P4 ⃰ᗘ᮲௳ୗ࡛ࡢⓎ⫱⎔ቃ㢮ఝࡋࡓࡶࡢ࡛࠶ࡿ⪃࠼ࡽࢀࡿࠋࡑࡢࡓࡵ㸪⮬↛Ⓨ⫱᮲ ௳ୗ࡛ࡢ Day 6 ࠾ࡅࡿ⾑୰ P4 ⃰ᗘࡣ㸪W1DF ࡁ࡞ᙳ㡪ࢆ࠼࡞࠸᥎ ࡉࢀࡓࠋ Ginther ࡽ㸦42㸧ࡣᮍ⤒⏘∵ࢆ⏝࠸ࡓヨ㦂࠾࠸࡚㸪㯤యྠ୍༸ᕢෆ࡛ඹᏑࡍࡿ W1DF ࡣᑐഃࡢ༸ᕢᏑᅾࡍࡿ W1DF ẚ㍑ࡋ࡚ᖹᆒ┤ᚄࡀᑠࡉ࠸ࡇࢆሗ࿌ࡋ࡚࠸ࡿࠋ⤒ ⏘㠀Ἢங∵ࢆ⏝࠸ࡓᮏヨ㦂࠾࠸࡚ࡶ㸪ྠᵝࡢ⤖ᯝࡀᚓࡽࢀࡓࠋ࣮࢝ࣛࢻࣉࣛࢆ⏝࠸ࡓ 64 ◊✲࡛㸪㯤య W1DF ࡀྠ୍༸ᕢෆ࡛Ⓨ⫱ࡋࡓሙྜ㸪W1DF ࡀ㯤యᑐഃࡢ༸ᕢ࡛Ⓨ ⫱ࡋࡓሙྜẚ㸪༸ᕢෆࡢ⾑⟶ᢠ್ࡀపୗࡋ࡚࠾ࡾ㸪W1DF ࡢ༸⬊ቨ⾑ὶࡀ㧗࠸ࡇ ࡀ♧ࡉࢀ࡚࠸ࡿ㸦42㸧 ࠋᮏヨ㦂࠾࠸࡚ࡶྠᵝࡢ⌧㇟ࡀ⏕ࡌ࡚࠸ࡓ᥎ ࡉࢀࡿࠋ㐣ཤ ࡢሗ࿌࠾࠸࡚㸪༸⬊ቨ⾑ὶࡣ㸪༸⬊ࡢᶵ⬟ࢆホ౯ࡍࡿࡢ᭷⏝࡛࠶ࡿࡉࢀ࡚࠸ࡿࡓ ࡵ㸦2㸪42㸪84㸪86㸧 㸪ඹᏑ⩌ࡢ W1DF ࡼࡾከࡃ⾑ᾮࡀ౪⤥ࡉࢀࡿࡇࡣ㸪㓟⣲㸪ᰤ㣴 ࡉࡽࡣ LH ࡞ࡢᛶ࣍ࣝࣔࣥࡢ౪⤥㔞㸦109㸧ࢆቑࡸࡋ㸪ࡑࡢ⤖ᯝ㸪༸⬊ෆࡢ⣽⬊ࡢቑ Ṫࡸ࣍ࣝࣔࣥ⏘⏕ࡀቑຍࡋ࡚㸪ᖍ༸⬊ࡢⓎ⫱ࢆಁ㐍ࡋࡓ⪃࠼ࡽࢀࡿࠋ୍᪉࡛㸪ᮏヨ 㦂࡛ࡣ㸪Day 6 ࠾ࡅࡿ⾑୰ P4 ⃰ᗘࢆඹᏑ⩌㠀ඹᏑ⩌࡛ẚ㍑ࡋࡓࡇࢁ㸪㠀ඹᏑ⩌࡛ ᭷ព㧗࠸ࡇࡀ♧ࡉࢀࡓࠋࡇࡢ⤖ᯝࢆࡶࡓࡽࡋࡓཎᅉࡣ࡛᫂࠶ࡾ㸪ᮏヨ㦂࡛ࡣ㯤య ࡢᖹᆒ┤ᚄࡢࢹ࣮ࢱࢆ♧ࡋ࡚࠸࡞࠸ࡓࡵ㸪㯤యࡢⓎ⫱ᕪ␗ࡀ⏕ࡌࡓࡇࡼࡿ⤖ᯝ࡛ ࠶ࡿࡢ㸪W1DF ࡀ༸ᕢෆᏑᅾࡍࡿࡇ࡛㯤యᶵ⬟㈇ࡢᙳ㡪ࢆ࠼ࡓ⤖ᯝ࡞ࡢ㸪 ᫂ࡽ࡛ࡁ࡞ࡗࡓࠋ ࡑࡇ࡛㸪ヨ㦂 2 ࡛ࡣ㸪ඹᏑ⩌㠀ඹᏑ⩌࡛ࡢ W1DF ࡢᛶ㉁ࢆヲ⣽᳨ウࡍࡿࡓࡵ㸪⤒ ⏘Ἢங∵ࢆ⏝࠸࡚㸪ඹᏑ⩌㠀ඹᏑ⩌ࡢ W1DF 㯤యࡢⓎ⫱ືែ㸪⾑୰ P4 ⃰ᗘ࠾ࡼࡧ༸ ⬊ᾮࢫࢸࣟࢻ࣍ࣝࣔࣥ⃰ᗘࢆẚ㍑ࡋࡓࠋヨ㦂 2 ࡛ࡣ㸪Day 0 ࡽ 4 ࡲ࡛ࡢ㯤యࡢⓎ⫱ ⾑୰ P4 ⃰ᗘ㛵ࡋ࡚ࡣ㸪 ືែ࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ࡣඹᏑ⩌㠀ඹᏑ⩌㛫࡛ᕪࡣ࡞ࡃ㸪 ヨ㦂 1 ྠᵝࡢ⤖ᯝࡣᚓࡽࢀ࡞ࡗࡓࠋࡲࡓ㸪W1DF ࡢᖹᆒ┤ᚄ᥎⛣࠾࠸࡚ࡶඹᏑ⩌ ࡛㧗ࡵࡢ᥎⛣ࢆ♧ࡋ࡚࠸ࡓࡀ㸪᭷ព࡞ᕪࡣࡳࡽࢀ࡞ࡗࡓࠋࡇࡢཎᅉࡋ࡚㸪ᮏヨ㦂࡛ ࡣ༸⬊ἼⓎ⌧ᚋ 5 ᪥┠ࡲ࡛ࡋືែࢆ㏣㊧ࡋ࡚࠾ࡽࡎ㸪ヨ㦂 1 ࡛ࡣ 7 ᪥┠↔Ⅼࢆᙜ࡚ ࡚࠸ࡓࡓࡵ㸪ྠᵝࡢ⤖ᯝࡀᚓࡽࢀ࡞ࡗࡓ⪃࠼ࡽࢀࡿࠋDay 4 ࡛ࡢ W1DF ࡢ༸⬊ᾮ୰ ࢫࢸࣟࢻ࣍ࣝࣔࣥࢆ ᐃࡋࡓࡇࢁ㸪ඹᏑ⩌࡛ P4 ⃰ᗘࡀ᭷ព㧗್ࢆ♧ࡋࡓࠋࡋࡋ ࡞ࡀࡽ㸪༸⬊ᾮ E2 ⃰ᗘࡸ A4 ⃰ᗘ᭷ព࡞ᕪࡣࡳࡽࢀ࡞ࡗࡓࠋ༸⬊ෆ࡛ࢫࢸࣟࢻ࣍ ࣝࣔࣥࡀྜᡂࡉࢀࡿ㐣⛬ࡣ㸪༸⬊ቨࡢෆ༸⬊⭷⣽⬊࠾࠸࡚㸪ࢫࢸࣟࢻ࣍ࣝࣔࣥࡢᇶ ㉁࡞ࡿࢥࣞࢫࢸ࣮ࣟࣝࡀ⣽⬊ෆྲྀࡾ㎸ࡲࢀࡿࡇࡽጞࡲࡾ㸪ࡇࡢࢥࣞࢫࢸ࣮ࣟࣝ 65 ࡣࣉࣞࢢࢿࣀࣟࣥኚࡉࢀ㸪ࡑࡢᚋࣉࣞࢢࢿࣀࣟࣥࡽ P4 ኚࡉࢀࡿ㸦22㸧 ࠋࡑ ࠋࡉࡽ A4 ࡣ༸⬊⭷⣽⬊ࡽ㢛 ࡢᚋ㸪༸⬊⭷⣽⬊ෆ࡛ P4 ࡣ A4 ኚࡉࢀࡿ㸦31㸪59㸧 ⢏ᒙ⣽⬊⛣⾜ࡋࡑࡇ࡛ E2 ኚࡉࢀࡿ㸦12㸧ࡇ࡛༸⬊ෆ࡛ࢫࢸࣟࢻ࣍ࣝࣔࣥࡀ⏘ ⏕ࡉࢀ࡚࠸ࡿࠋᅇࡢ⤖ᯝ࡛ࡣ㸪A4 ࠾ࡼࡧ E2 ⃰ᗘᕪࡣࡳࡽࢀ࡞ࡗࡓࡇࡽ㸪ඹᏑ ⩌࠾ࡼࡧ㠀ඹᏑ⩌ࡑࢀࡒࢀࡢᖍ༸⬊ࡢ༸⬊⭷⣽⬊࠾ࡼࡧ㢛⢏ᒙ⣽⬊ࡢࢫࢸࣟࢻ࣍ࣝ ࣔࣥ⏘⏕⬟ᕪࡣ࡞࠸ࡇࡀ᥎ ࡉࢀࡿࠋࡑࡢࡓࡵ㸪༸⬊ᾮ P4 ⃰ᗘࡀඹᏑ⩌࡛㧗࠸⌮⏤ ࡋ࡚㸪༸⬊࡛ࡢ P4 ⏘⏕⬟ࡀ㧗ࡗࡓࡓࡵ᥎ ࡍࡿࡇࡣ㞴ࡋ࠸ࠋ㐣ཤࡢሗ࿌࡛㸪༸ ᕢෆ࡛ࡢ୍ࡘࡢື⬦ࡽ㯤యࡑࡢ㏆㞄ࡢ༸⬊ື⬦ᯞࢆఙࡤࡋ࡚࠸ࡿྍ⬟ᛶࡀ㸪ࣄࢺ ࡢ༸ᕢࢆ⏝࠸ࡓ◊✲࡛♧ࡉࢀ࡚࠸ࡿ㸦75㸧 ࠋࡇࡢࡇࡣ㸪㯤యࡽࡢ⾑⟶ࡀྠ୍༸ᕢෆ Ꮡᅾࡍࡿ༸⬊ቨᏑᅾࡍࡿẟ⣽⾑⟶࿀ྜࡍࡿࡇ࡛㸪㯤యࡽᨺฟࡉࢀࡓ㧗⃰ᗘࡢ P4 ࡀ༸⬊౪⤥ࡉࢀ㸪༸⬊ᾮෆᾐ㏱ࡋ࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋᮏヨ㦂࡛ࡣ㸪ࡑࡢ௬ ㄝࢆド᫂ࡍࡿࢹ࣮ࢱࢆ♧ࡋ࡚࠸࡞࠸ࡀ㸪ᮏヨ㦂࡛ᚓࡽࢀࡓ⤖ᯝࡣ㸪ࡇࡢ௬ㄝࡢྍ⬟ᛶࢆ ♧၀ࡍࡿࡶࡢ⪃࠼ࡽࢀࡿࠋࡲࡓ㸪⾑୰ E2 ⃰ᗘࡣඹᏑ⩌࡛㧗࠸ഴྥࢆ♧ࡋࡓࠋࡇࢀࡣ㸪 Ginther ࡽ㸦42㸧ࡀᮍ⤒⏘∵ࢆ⏝࠸࡚⾜ࡗࡓ◊✲࡛ሗ࿌ࡋ࡚࠸ࡿࡼ࠺㸪㯤య W1DF ࡀྠ୍༸ᕢෆ࡛ඹᏑࡍࡿሙྜ㸪W1DF ࡢ༸⬊ቨ⾑ὶࡀ㠀ඹᏑࡢሙྜẚ࡚ࡁ࠸ࡇ ࡽ㸪ᮏヨ㦂࠾࠸࡚ࡶ㸪ඹᏑ⩌࡛ࡢ W1DF ࡢ⾑ᾮ㑏ὶࡀࡼࡾࡁࡃ㸪ᖍ༸⬊ෆ࡛ ⏘⏕ࡉࢀࡓ E2 ࡢᨺฟࡀஹ㐍ࡋࡓࡇࡀཎᅉ⪃࠼ࡽࢀࡿࠋࡋࡋ࡞ࡀࡽ㸪ᮏヨ㦂࡛ࡣ༸ ⬊ቨ⾑ὶࡢホ౯ࡣ⾜ࡗ࡚࠾ࡽࡎ㸪༑ศ᳨ド࡛ࡁࡿࢹ࣮ࢱࡣ࡞࠸ࠋ㠀Ἢங⤒⏘∵ࢆ⏝࠸ ࡚ࡢ༸⬊ቨ⾑ὶホ౯ࢆ⾜࠸㸪ࡇࡢཎᅉࢆ᫂ࡽࡍࡿ◊✲ࢆ⾜ࡗ࡚࠸ࡃᚲせࡀ࠶ࡿ⪃ ࠼ࡿࠋ 3㸫6㸬ᑠᣓ ᮏᐇ㦂࡛ࡣ㸪㠀Ἢங⤒⏘∵ࢆ⏝࠸࡚ W1DF ࡢⓎ⫱ࡸᛶ㉁ࢆไᚚࡍࡿせᅉ㸪ࡲࡓඹᏑ⩌ 㠀ඹᏑ⩌࡛ࡢ W1DF ࡢᛶ㉁ࡘ࠸࡚ㄪࡓࠋࡑࡢ⤖ᯝ㸪ヨ㦂 1 ࠾࠸࡚༸☜ㄆ᪥ 66 ࡽ 7 ᪥┠ࡢ W1DF ࡢᖹᆒ┤ᚄ⾑୰ P4 ⃰ᗘࡢ㛫┦㛵㛵ಀࡣ࡞࠸ࡇࡀ♧ࡉࢀࡓࡀ㸪 W1DF ࡀ㯤యྠ୍༸ᕢෆ࡛ඹᏑࡍࡿ㸪ඹᏑࡋ࡞࠸ W1DF ẚ࡚༸☜ㄆ᪥ࡽ㸵 ᪥┠ࡢᖹᆒ┤ᚄࡀࡁࡃ࡞ࡾ㸪⾑୰ P4 ⃰ᗘࡶ㧗್ࢆ♧ࡍࡇࡀ♧ࡉࢀࡓࠋࡇࢀࡽࡢࡇ ࡽ㸪W1DF ࡢᙧែࡣ⾑୰ P4 ⃰ᗘ࡛ࡣ࡞ࡃ㸪༸ᕢෆ࡛ࡢ㯤యࡢ⨨㛵ಀࡀࡁࡃ㛵 ࡍࡿࡇࡀ♧၀ࡉࢀࡓࠋヨ㦂 2 ࠾࠸࡚㸪༸☜ㄆ᪥ࡽ 5 ᪥┠ࡲ࡛ࡢᖍ༸⬊ࡢᖹ ᆒ┤ᚄ࠾ࡼࡧ⾑୰ P4 ⃰ᗘඹᏑ⩌㠀ඹᏑ⩌ࡢ㛫࡛ᕪࡣࡳࡽࢀ࡞ࡗࡓࡀ㸪༸⬊ᾮ୰ ࡢ P4 ⃰ᗘࡀඹᏑ⩌࡛㧗࠸࠸࠺⤖ᯝࡀᚓࡽࢀࡓࠋࡇࡢ⌧㇟ࡀ⏕ࡌࡓཎᅉࡋ࡚㸪༸ᕢෆ ࡛ࡢ㏆㞄ࡢ㯤యࡽ⏘⏕ࡉࢀࡓ P4 ࡀ⾑⟶ࢆ㏻ࡌ࡚ᖍ༸⬊ࡢ༸⬊ቨẟ⣽⾑⟶ධࡋ㸪 P4 ࡀ༸⬊ᾮෆᾐ㏱ࡋࡓ࠸࠺ྍ⬟ᛶࡶ⪃࠼ࡽࢀࡿࡀ㸪༸ᕢෆࡢ⾑⟶㉮⾜ࡸᙧᡂࡢ≧ἣ ࡢゎ᫂ࡶྵࡵ࡚᭦࡞ࡿ◊✲ࢆ⾜ࡗ࡚࠸ࡃᚲせࡀ࠶ࡿ⪃࠼ࡽࢀࡿࠋ 67 Table 3.1. Plasma P4 concentration on Day 6. P4 (ng/ml) IG CG P-value 2.7 㼼 0.3 3.5 㼼 0.3 P < 0.05 Values are mean 㼼 SEM. IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL. Day0 = The day of the first follicular wave emergence. 68 Plasma P4 concentration (ng/ml) 7.0 R=0.23 P:NS 6.0 5.0 4.0 3.0 2.0 1.0 13 14 15 16 17 18 Diameter of the first wave dominant follicle (mm) Fig. 3.1. The linear relationship between plasma P4 concentration and diameter of the first wave dominant follicle on Day 6. Day 0 = The day of the first follicular wave emergence. 69 Plasma P4 concentration (ng/ml) 7.0 IG R=0.36 P:NS CG R=0.12 P:NS 6.0 5.0 4.0 3.0 2.0 1.0 13 14 15 16 17 18 Diameter of the first wave dominant follicle (mm) Fig. 3.2. The linear relationship between plasma P4 concentration and diameter of the first wave dominant follicle (DF) on Day6 in IG and CG. Day0 = The day of the first follicular wave emergence. IG; Ipsilateral group: DF ipsilateral to the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL. 70 Table 3.2. Diameter of the first follicular wave dominant follicle on Day6. Diameter (mm) IG CG P-value 15.5 㼼 0.2 14.7 㼼 0.2 P < 0.01 Values are mean 㼼 SEM. IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL. Day0 = The day of the first follicular wave emergence. 71 Diameter of dominant follicle (mm) 14 IG CG 12 10 8 6 G : NS 4 D : P < 0.001 G,D : NS 2 0 Day0 Day1 Day2 Day3 Day4 Days from follicular wave emergence Fig. 3.3. Comparative changes of the first follicular wave dominant follicle (DF) of IG and CG from Day 0 to 4. Day 0 = The day of the first follicular wave emergence. Effect of group (G) (NS), day (D) (P < 0.01) and interaction between group and day (G,D) (NS). IG; Ipsilateral group: DF ipsilateral to the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL. Values are mean r SEM of each point. 72 Cross-sectional area of corpus luteum (mm2) 300 IG CG 250 200 150 G : NS 100 D : P < 0.001 50 0 G,D : NS Day0 Day1 Day2 Day3 Day4 Days wave emergence Daysfrom fromfollicular follicular wave emergence Fig. 3.4. Comparative changes of cross-sectional area of corpus luteum (CL) of IG and CG from Day 0 to 4. Day 0 = The day of the first follicular wave emergence. Effect of group (G) (NS), day (D) (P < 0.001) and interaction between group and day (G,D) (NS). IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the CL. CG; Contralateral group: DF contralateral to the CL. Values are mean r SEM of each point. 73 Plasma P4 concentration (ng/ml) 3.0 2.5 2.0 IG CG 1.5 1.0 G : NS D : P < 0.001 G,D : NS 0.5 0 Day0 Day1 Day2 Day3 Day4 Days from follicular wave emergence Fig. 3.5. Comparative changes of plasma P4 concentration of IG and CG from Day 0 to 4. Day 0 = The day of the first follicular wave emergence. Effect of group (G) (NS), day (D) (P<0.001) and interaction between group and day (G,D) (NS). IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL. Values are mean r SEM of each point. 74 Table 3.3. Plasma E2 concentration on Day4. E2 (pg/ml) IG CG P-value 1.69 㼼 0.12 1.51 㼼 0.15 P < 0.1 Values are mean 㼼 SEM. IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL. Day0 = The day of the first follicular wave emergence. 75 Table 3.4. Sex steroids concentration in follicular fluid on Day4. IG CG P-value P4 (ng/ml) 48.8 㼼 12.7 16.2 㼼 3.2 P < 0.05 E2 (ng/ml) 419.2 㼼 33.2 339.4 㼼 53.5 NS A4 (ng/ml) 84.6 㼼 23.1 102.7 㼼 40.2 NS Values are mean 㼼 SEM. IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG; Contralateral group: DF contralateral to the CL. Day0 = The day of the first follicular wave emergence. 76 ➨㸲❶ 㯤యྠ୍༸ᕢෆᏑᅾࡍࡿ➨ 1 ༸⬊Ἴᖍ༸⬊㸦W1DF㸧 ࡢᛶ㉁࠾ࡼࡧཷ⫾ᛶཬࡰࡍᙳ㡪 77 4㸫1㸬ᗎㄽ ➨ 3 ❶ࡼࡾ㸪W1DF ࡢⓎ⫱ไᚚ༸ᕢෆ࡛ࡢ㯤యࡢඹᏑࡢ᭷↓ࡀ㔜せ࡛࠶ࡿࡇࡀ ᫂ࡽ࡞ࡗࡓࠋࡑࡇ࡛㸪ᮏ❶࡛ࡣࡇࡢ⌧㇟ࡀἪங∵࠾࠸࡚ࡶྠᵝ㉳ࡇࡿ⌧㇟࡛࠶ ࡿࡢࢆ᫂ࡽࡍࡿࡓࡵ㸪W1DF 㯤యࡢ⨨㛵ಀࡀ W1DF ࠼ࡿᙳ㡪ࢆㄪ ࡓࠋࡇࢀࡲ࡛㸪㯤యࡀᏑᅾࡍࡿ༸ᕢྠഃࡢᏊᐑゅ࠾ࡅࡿᏊᐑෆ⭷ࡢ⤌⧊୰ P4 ⃰ᗘࡣ㸪 ᑐഃࡢᏊᐑゅ࠾ࡅࡿᏊᐑෆ⭷ẚ㧗࠸ࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸦20㸧 ࠋࡲࡓ㸪༸๓ ༸⬊ࡀᏑᅾࡍࡿ༸ᕢྠഃࡢ༸⟶ෆ E2 ⃰ᗘࡣ㸪ᑐഃࡢࡶࡢẚ㍑ࡋ࡚㧗࠸ࡇࡀሗ࿌ ࡉࢀ࡚࠸ࡿ㸦99㸧 ࠋࡇࢀࡽࡢᐇࡽ㸪༸ᕢྠഃࡢᏊᐑゅࡢ㛫ࡣᐦ᥋࡞ᒁᡤࡢࡘ࡞ ࡀࡾࡀᏑᅾࡋ㸪Ꮚᐑࡀྠഃࡢ༸ᕢࡽศἪࡉࢀࡿᛶࢫࢸࣟࢻ࣍ࣝࣔࣥࡢᙳ㡪ࢆᙉࡃཷ ࡅ࡚࠸ࡿྍ⬟ᛶࡀ♧၀ࡉࢀࡿࠋࡑࡇ࡛㸪ேᕤᤵ⢭ᚋࡢཷ⫾⋡ࡘ࠸࡚㸪ඹᏑ⩌㠀ඹᏑ ⩌࡛ẚ㍑ࡋ㸪W1DF 㯤యࡢ⨨㛵ಀࡀ࢘ࢩࡢ⦾Ṫ⏕⌮࠼ࡿᙳ㡪ࢆㄪᰝࡋࡓࠋ 4㸫2㸬ヨ㦂 1㸸Ἢங∵࠾ࡅࡿ W1DF 㯤యࡢඹᏑ࠾ࡼࡧ㠀ඹᏑ㛵ಀࡀ W1DF 㯤యࡢ Ⓨ⫱ືែ࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ཬࡰࡍᙳ㡪 4㸫2㸫1㸬౪ヨື≀ ᖏᗈ␆⏘Ꮫ␆⏘ࣇ࣮ࣝࢻ⛉Ꮫࢭࣥࢱ࣮࡚㣫㣴ࡉࢀ࡚࠸ࡿ㸪⮫ᗋⓗᗣ࡛Ⓨ ࿘ᮇࡀṇᖖ࡞࣍ࣝࢫࢱࣥ✀⤒⏘Ἢங∵㸦n㸻30㸪ᖹᆒ⏘ḟ㸸2.2 r 0.2㸪ᖹᆒศፔᚋ᪥ᩘ㸸 99.9s8.0㸪ᖹᆒ᪥ங㔞㸸37.6s1.6 kg㸧ࢆ⏝࠸ࡓࠋ౪ヨࡋࡓಶయࡣ㸪ศፔᚋ௦ㅰᛶᝈ ࡸ⦾Ṫ㞀ᐖ࡛⒪Ṕࡢ࡞࠸ࡶࡢ࡛࠶ࡿࠋ 4㸫2㸫2㸬ᐇ㦂᪉ἲ ⮬↛ⓎⓎぢᚋ㸪24 㛫ẖ┤⭠᳨ᰝࡲࡓࡣ㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚༸ࡢ☜ㄆ ࢆ⾜ࡗࡓࠋ༸☜ㄆ᪥ࢆ Day 0 ࡋࡓࠋDay 2㸪4㸪6㸪8 ࠾ࡼࡧ 10 ࡢ㛫㉸㡢Ἴ⏬ീデ᩿ 78 ⨨ࢆ⏝࠸࡚༸ᕢࡢほᐹࢆ⾜ࡗࡓࠋࡇࡢ㸪ᖍ༸⬊ࡀ 2 ࡘ࠶ࡿಶయ㸪ࡲࡓࡣᖍ༸⬊ ࡀ☜ㄆࡉࢀ࡞࠸ಶయࡣヨ㦂ࡼࡾ㝖እࡋࡓࠋࡲࡓ㸪ྠࡌ᪥ṇ୰ᑿ㟼⬦ࡼࡾ᥇⾑ࢆ⾜ࡗࡓࠋ 4㸫2㸫3㸬 ༸ᕢࡢほᐹ W1DF 㯤యࡢඹᏑࡲࡓࡣ㠀ඹᏑ㛵ಀࡢほᐹࡣ㸪5.0MHz ࡢࣜࢽࣉ࣮ࣟࣈࡀᒓ ࡋࡓ⤒┤⭠㉸㡢Ἴ⏬ീデ᩿⨨㸦HS̺101V㸪ᮏከ㟁Ꮚ㸪ឡ▱㸧ࢆ⏝࠸ࡓࠋ༸ᕢほᐹ ᭱┤ᚄࢆ᭷ࡍࡿ༸⬊ࢆ W1DF ࡋ㸪W1DF ࡀྠ୍༸ᕢෆ㯤యඹᏑࡍࡿሙྜࢆඹᏑ ⩌㸪㯤యࡀᏑᅾࡍࡿ༸ᕢࡣᑐഃࡢ༸ᕢᏑᅾࡍࡿሙྜࢆ㠀ඹᏑ⩌ࡋࡓࠋ 4㸫2㸫4㸬 ༸⬊࠾ࡼࡧ㯤యᙧែࡢほᐹ ᖍ༸⬊࠾ࡼࡧ㯤యࡢ᩿㠃✚ࡣ㸪5.0MHzࡢࣜࢽࣉ࣮ࣟࣈࡀᒓࡋࡓ⤒┤⭠㉸㡢Ἴ⏬ ീデ᩿⨨㸦HS̺101V㸪ᮏከ㟁Ꮚ㸪ឡ▱㸧ࢆ⏝࠸࡚ ᐃࡋࡓࠋࣉ࣮ࣟࣈࢆᡭᣦࡶ ┤⭠ෆᤄධࡋ㸪ᕥྑ୧ഃ༸ᕢࡢ᩿ᒙീࢆほᐹࡋ㸪ᖍ༸⬊࠾ࡼࡧ㯤యࡢ᩿ᒙ⏬ീࡀ᭱ ࡞ࡿࡼ࠺㟼Ṇ⏬ࢆグ㘓ࡋ㸪ࣃࢯࢥࣥୖ㟼Ṇ⏬ࢆ⛣ືࡉࡏ㸪ᖍ༸⬊ࡣ㛗ᚄ▷ᚄࡢᖹ ᆒ್ࢆᖹᆒ┤ᚄࡋ㸪ࢹ࣮ࢱࡋ࡚⏝࠸ࡓࠋ㯤యࡢ᩿㠃✚ࡣ㸪㯤యࡢ᩿ᒙ⏬ീࡀ᭱࡞ ࡿࡼ࠺㟼Ṇ⏬ࢆグ㘓ࡋ㸪ࣃࢯࢥࣥୖ㟼Ṇ⏬ࢆ⛣ືࡉࡏ㸪㯤యࡢ㛗ᚄ▷ᚄࡢᖹᆒ್ࡽ ᖹᆒ┤ᚄࢆ⟬ฟࡋ㸪㸦ᖹᆒ┤ᚄ㸭2㸧23.14ᙜ࡚ࡣࡵ㯤య᩿㠃✚ࢆ⟬ฟࡋ㸪ࢹ࣮ࢱ⏝ ࠸ࡓࠋ 4㸫2㸫5㸬᥇⾑ 21G᥇⾑㔪࣊ࣃࣜࣥຍ┿✵᥇⾑⟶㸦࣋ࣀࢪ࢙ࢡࢺϩ┿✵᥇⾑⟶㸪TERUMO㸪ᮾி㸧ࢆ ⏝࠸࡚ᑿ㟼⬦ࡼࡾ10mlࡢ᥇⾑ࢆ⾜ࡗࡓࠋ᥇ྲྀࡋࡓ⾑ᾮࡣịỈ୰࡚ᛴ෭ᚋ㸪4Υ࡛2,000 g㸪 20ศ㛫㐲ᚰࡋ㸪⾑₢ศ㞳ᚋ㸪⾑୰P4⃰ᗘ ᐃࡲ࡛㸫30Υ࡚⤖ಖᏑࡋࡓࠋ 79 4㸫2㸫6㸬⾑୰P4⃰ᗘࡢ ᐃ Day 2㸪4㸪6㸪8࠾ࡼࡧ10࠾ࡅࡿ⾑୰P4⃰ᗘࡢ ᐃࡣEIAἲࡼࡾ ᐃࡋࡓࠋ ᐃ᪉ἲ ࡣ➨2❶ྠᵝࡢ᪉ἲ࡛⾜ࡗࡓࠋ⾑୰P4ࡢ ᐃෆኚື࠾ࡼࡧ ᐃ㛫ኚືࡣࡑࢀࡒࢀ5.2㸣࠾ ࡼࡧ7.4%࡛࠶ࡗࡓࠋ 4㸫2㸫7㸬⤫ィฎ⌮ ⤖ᯝࡣࡍ࡚ᖹᆒsᶆ‽ㄗᕪ࡛♧ࡋࡓࠋ⾑୰ P4 ⃰ᗘ㸪ᖍ༸⬊ࡢᖹᆒ┤ᚄ࠾ࡼࡧ㯤య ᩿㠃✚ࡢ᥎⛣ࡢẚ㍑ࡣ㸪2 ඖ㓄⨨ศᩓศᯒࢆ⾜࠸㸪యࡢኚࡢ᭷ពᛶࢆㄪ㸪 స⏝ࡀ᭷ព࡛࠶ࡗࡓࡶࡢᑐࡋ࡚㸪 Bartlet ᳨ᐃࢆ⾜࠸ศᩓࡢᆒ୍ᛶࢆ᳨ᐃࡋࡓᚋ㸪 Pos-hoc ᳨ᐃࡋ࡚ Sheffe ࡢከ⩌ẚ㍑᳨ᐃࢆ⾜ࡗࡓࠋP < 0.05 ࡢሙྜࢆ᭷ពᕪ࠶ࡾࡋࡓࠋ⤫ィ ฎ⌮ࡣ⤫ィฎ⌮ࢯࣇࢺ࡛࠶ࡿ StatView㸦Abacus Concepts㸪Inc.㸪Berkeley㸪CA㸪USA㸧 ࢆ⏝࠸ࡓࠋ 4㸫3㸬ヨ㦂 2㸸W1DF 㯤యࡢඹᏑ࠾ࡼࡧ㠀ඹᏑ㛵ಀࡀேᕤᤵ⢭ᚋࡢཷ⫾⋡ཬࡰࡍᙳ 㡪 4㸫3㸫1㸬౪ヨື≀ ᖏᗈ␆⏘Ꮫ␆⏘ࣇ࣮ࣝࢻ⛉Ꮫࢭࣥࢱ࣮࡚㣫㣴ࡉࢀ࡚࠸ࡿ㸪⮫ᗋⓗᗣ࡛Ⓨ ࿘ᮇࡀṇᖖ࡞࣍ࣝࢫࢱࣥ✀⤒⏘Ἢங∵㸦153 㢌㸧ᮍ⤒⏘∵㸦75 㢌㸧ࢆ⏝࠸ࡓࠋヨ㦂 ᮇ㛫ࡣ 2008 ᖺ 3 ᭶㹼2013 ᖺ 9 ᭶࡛࠶ࡿࠋ౪ヨࡋࡓಶయࡣ㸪ศፔᚋ௦ㅰᛶᝈࡸ⦾Ṫ 㞀ᐖ࡛⒪Ṕࡢ࠶ࡿಶయࢆ㝖࠸ࡓࠋゎᯒࡣ㸪ᘏ 238 㢌ࡢἪங∵㹙ᖹᆒศፔᚋ᪥ᩘ㸸 119.8 r 5.2 ᪥㸪ᖹᆒ⏘ḟ㸸2.2 r 0.1㸪ᖹᆒἪங㔞㸸33.9 r 0.5 kg/᪥㸪ᖹᆒ࣎ࢹ࣭ࢥࣥࢹ 80 ࢩ࣭ࣙࣥࢫࢥ㸦BCS㸧 㸸2.91 r 0.02㸪ᖹᆒయ㔜㸸637.4 r 4.6 kg; ᖹᆒ r ᶆ‽ㄗᕪ㹛࠾ ࡼࡧᘏ 112 㢌ࡢᮍ⤒⏘∵㸦ᖹᆒ᭶㱋㸸14.2 r 0.3 ࣨ᭶㱋㸪ᖹᆒయ㔜㸸440.6 r 5.0 kg)ࢆ౪ ヨࡋࡓࠋἪங∵࠾ࡼࡧᮍ⤒⏘∵ࡣࢥ࣮ࣥࢧ࣮ࣞࢪ㸪ࢢࣛࢫࢧ࣮ࣞࢪ㸪㇋⢑㸪ࢺ࢘ ࣔࣟࢥࢩ✐⢏࠾ࡼࡧ㓄ྜ㣫ᩱࢆΰྜࡋࡓ TMR ࢆ⤥ࡉࢀ࡚࠾ࡾ㸪⮬⏤㣧Ỉ࡛ࡁࡿࣇࣜ ࣮ࢫࢺ࣮࡚ࣝ㣫㣴ࡉࢀ࡚࠸ࡓࠋࡲࡓ㸪Ἢங∵ࡣ 1 ᪥ 2 ᅇᦢஙࡉࢀ࡚࠸ࡓࠋேᕤᤵ⢭ࡣ㸪 Ἢங∵࠾࠸࡚ࡣศፔᚋ 60 ᪥ࡽ㸪ᮍ⤒⏘∵࠾࠸࡚ࡣయ㔜ࡀ 350 ௨ୖ࡞ࡗࡓⅬ ࡽ㛤ጞࡉࢀࡓࠋ 4㸫3㸫2㸬ᐇ㦂᪉ἲ እ㒊Ⓨᚩೃ㸪ࢸ࣮ࣝ࣌ࣥࢺᾘኻࡢ᭷↓㸪⭐㙾᳨ᰝ࠾ࡼࡧ┤⭠᳨ᰝࡼࡿ༸๓༸ ⬊ࡢᏑᅾࡸ㏥⾜㯤యࡢ☜ㄆࡼࡿ⮬↛ⓎⓎぢᚋேᕤᤵ⢭ࢆᐇࡋࡓࠋேᕤᤵ⢭ᐇ 24 㛫ᚋ┤⭠᳨ᰝࡲࡓࡣ㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚༸ࡢ☜ㄆࢆ⾜ࡗࡓࠋ༸ࡋ࡚ ࠸࡞࠸ሙྜ㸪ᗘேᕤᤵ⢭ࢆᐇࡋ㸪24 㛫ᚋᗘ༸☜ㄆࢆ⾜ࡗࡓࠋࡇࡢ㸪2 ᅇ ࡢேᕤᤵ⢭ᐇᚋ㸪༸ࡋ࡚࠸࡞ࡗࡓಶయࡣヨ㦂ࡼࡾ㝖እࡋࡓࠋ༸☜ㄆ᪥ࢆ Day 0 ࡋ㸪Day 4 ࡽ 8 ࡢ㛫┤⭠᳨ᰝࢆ⾜࠸㸪W1DF ࠾ࡼࡧ㯤యࡢ⨨㛵ಀࢆ㉸㡢Ἴ⏬ീ デ᩿⨨ࢆ⏝࠸࡚☜ㄆࡋࡓࠋࡇࡢᖍ༸⬊ࡀ 2 ࡘ࠶ࡿಶయ㸪ࡲࡓࡣᖍ༸⬊ࡀ☜ㄆࡉ ࢀ࡞࠸ಶయࡣヨ㦂ࡼࡾ㝖እࡋࡓࠋேᕤᤵ⢭ᚋ 40 ᪥┠㉸㡢Ἴ⏬ീデ᩿⨨ࢆ⏝࠸࡚ዷፎ デ᩿ࢆ⾜࠸㸪ཷ⫾⋡ࢆ⟬ฟࡋࡓࠋ 4㸫3㸫3㸬 ༸ᕢࡢほᐹ W1DF 㯤యࡢඹᏑ࣭㠀ඹᏑ㛵ಀࡢほᐹࡣ㸪5.0MHz ࡢࣜࢽࣉ࣮ࣟࣈࡀᒓࡋࡓ ⤒┤⭠㉸㡢Ἴ⏬ീデ᩿⨨㸦HS̺101V㸪ᮏከ㟁Ꮚ㸪ឡ▱㸧ࢆ⏝࠸ࡓࠋ༸ᕢほᐹ᭱ ┤ᚄࢆ᭷ࡍࡿ༸⬊ࢆ W1DF ࡋ㸪W1DF ࡀྠ୍༸ᕢෆ㯤యඹᏑࡍࡿሙྜࢆඹᏑ⩌㸪 81 㯤యࡀᏑᅾࡍࡿ༸ᕢࡣᑐഃࡢ༸ᕢᏑᅾࡍࡿሙྜࢆ㠀ඹᏑ⩌ࡋࡓࠋ 4㸫3㸫4㸬ཷ⫾⋡ࡢ⟬ฟ ேᕤᤵ⢭ࡋࡓᘏ㢌ᩘᑐࡍࡿ㸪ࡑࡢேᕤᤵ⢭࡛ዷፎࡋࡓ㢌ᩘࡢྜࡋ࡚㸪ཷ⫾⋡ ࢆ⟬ฟࡋࡓࠋ 4㸫3㸫5㸬⤫ィฎ⌮ ཷ⫾⋡ࡢホ౯ࡣȮ᳨ᐃ࠾ࡼࡧࣟࢪࢫࢸࢵࢡᅇᖐศᯒࢆ⏝࠸ࡓࠋ࡚ࡢ࢘ࢩ ᑐࡍࡿཷ⫾⋡ࡢホ౯ࡣȮ᳨ᐃࢆ⏝࠸ࡓࠋ⤒⏘Ἢங∵ᮍ⤒⏘∵ࡢཷ⫾⋡ཬࡰࡍ ᙳ㡪ࢆヲ⣽ホ౯ࡍࡿࡓࡵ㸪ࣟࢪࢫࢸࢵࢡᅇᖐศᯒࢆ⏝࠸ࡓࠋ⤒⏘∵ᮍ⤒⏘∵ ࡶཷ⫾⋡ࢆᚑᒓኚᩘࡋࡓࠋ⤒⏘∵ᮍ⤒⏘∵ඹ㏻ࡢ⊂❧ኚᩘࡣ㸪ྠ୍༸ᕢෆ࡛ ࡢ㯤య W1DF ࡢඹᏑࡢ᭷↓㸦ඹᏑ㸪㠀ඹᏑ㸧 㸪Ꮨ⠇㸦 ᬮ㸸6㹼9 ᭶㸪෭ᾴ㸸10㹼5 ᭶㸧 ࡛࠶ࡿࠋຍ࠼࡚㸪ᮍ⤒⏘∵࡛ࡣయ㔜㸦㸺441kg㸪Ӎ441kg㸧ࢆ⊂❧ኚᩘࡋ࡚㸪ࣟࢪࢫࢸ ࢵࢡᅇᖐศᯒࢆ⾜ࡗࡓࠋ⤒⏘Ἢங∵࡛ࡣศፔᚋ᪥ᩘ㸦㸺120 ᪥㸪Ӎ120 ᪥㸧 㸪Ἢங㔞㸦㸺 34kg/᪥㸪Ӎ34kg/᪥㸧 㸪⏘ḟ㸦1㸪2㸪Ӎ3㸧 㸪࣎ࢹ࣭ࢥࣥࢹࢩ࣭ࣙࣥࢫࢥ㸦BCS㸹㸺3㸪 Ӎ3㸧 㸦27㸧࠾ࡼࡧయ㔜㸦㸺637kg㸪Ӎ637kg㸧ࢆ⊂❧ኚᩘ㏣ຍࡋ࡚ࣟࢪࢫࢸࢵࢡᅇᖐ ศᯒࢆ⾜ࡗࡓࠋἪங㔞ࡣ᭶ 1 ᅇ⾜ࢃࢀࡿ∵⩌᳨ᐃࡢࢹ࣮ࢱࢆ⏝ࡋ㸪BCS ࠾ࡼࡧయ㔜ࡣ ␆⏘ࣇ࣮ࣝࢻ⛉Ꮫࢭࣥࢱ࣮࡛ẖ᭶⾜ࢃࢀࡿ ᐃ࡛ᚓࡽࢀࡓࡶࡢࡢ࠺ࡕ㸪ேᕤᤵ⢭ࡽ ┤㏆࡛ᚓࡽࢀࡓࢹ࣮ࢱࢆ⏝ࡋࡓࠋἪங㔞㸪BCS ࠾ࡼࡧయ㔜ࡣṇつศᕸᚑࡗ࡚࠸ࡓࡓ ࡵ㸪ᖹᆒ್࡛ 2 ⩌ศࡅࢡࣛࢫศࡅࢆ⾜ࡗࡓࠋP < 0.05 ࢆ᭷ពᕪ࠶ࡾࡋࡓࠋ࡚ࡢ᳨ ᐃࡣ R ࢯࣇࢺ࢙࢘㸦version 2.14.0 for Mac OS X.㸧ࢆ⏝࠸ࡓࠋ 4㸫4㸬⤖ᯝ 82 4㸫4㸫1㸬ヨ㦂 1 ⤖ᯝ ᮏヨ㦂࡛ඹᏑ⩌ࡣ n㸻16㸦ᖹᆒศፔᚋ᪥ᩘ㸸104.8s10.5㸪ᖹᆒ⏘ḟ㸸1.9s0.2㸪ᖹᆒἪ ங㔞㸸35.5s2.1 kg/᪥㸧 㸪㠀ඹᏑ⩌ࡣ n㸻14㸦ᖹᆒศፔᚋ᪥ᩘ㸸94.4s12.4㸪ᖹᆒ⏘ḟ㸸2.6 s0.4㸪ᖹᆒἪங㔞㸸39.9s2.3 kg/᪥㸧࡛࠶ࡗࡓࠋ㯤యࡢ᩿㠃✚ࡢ᥎⛣ࢆ Fig. 4.1.♧ࡋࡓࠋ ඹᏑ⩌㠀ඹᏑ⩌㛫࡛㯤య᩿㠃✚ࡢ᥎⛣ᕪࡣ࡞ࡗࡓࠋᖍ༸⬊ࡢᖹᆒ┤ᚄࡢ᥎⛣ࢆ Fig. 4.2.♧ࡋࡓࠋDay 8 ࡛ඹᏑ⩌ࡣ㠀ඹᏑ⩌ẚ㍑ࡋ࡚᭷ពࡁࡃⓎ⫱ࡋ࡚࠸ࡓࠋࡲ ࡓ㸪ࡑࢀࡒࢀࡢ⩌ෆ࡛ࡢ᥎⛣ࢆẚ㍑ࡋࡓࡇࢁ㸪ඹᏑ⩌࡛ࡣ Day 4 ࡽ 6 ࠾ࡼࡧ 8 ࡅ࡚᭷ពࡁࡃⓎ⫱ࡋࡓࡀ㸪㠀ඹᏑ⩌࡛ࡣ Day 4 ࡽ 8 ࡅ࡚ࡣࡌࡵ࡚᭷ពࡁ ࡃⓎ⫱ࡋࡓࠋඹᏑ⩌㠀ඹᏑ⩌࡛⾑୰ P4 ⃰ᗘࡢ᥎⛣ࢆẚ㍑ࡋࡓࡇࢁ㸪ᕪࡣࡳࡽࢀ࡞ ࡗࡓ㸦Fig. 4.3㸧 ࠋ 4㸫4㸫2㸬ヨ㦂 2 ⤖ᯝ ᮏヨ㦂࡛㸪౪ヨ∵࡚ࡢ⥲ཷ⫾⋡ࡣ 54.0㸣㸦189/350㸧 㸪⤒⏘∵࡛ࡢ⥲ཷ⫾⋡ࡣ 48.9㸣 㸦122/238㸧 㸪ᮍ⤒⏘∵࡛ࡢ⥲ཷ⫾⋡ࡣ 58.9㸣㸦67/112㸧࡛࠶ࡗࡓࠋ ᮏヨ㦂࡛⤒⏘∵ᮍ⤒⏘∵ࢆྜࢃࡏࡓࡁࡢඹᏑ⩌ࡣ n㸻184㸪㠀ඹᏑ⩌ࡣ n㸻166 ࡛ ࠶ࡗࡓࠋ⤒⏘∵࠾ࡅࡿඹᏑ⩌ࡣ n㸻129㸦ᖹᆒศፔᚋ᪥ᩘ㸸115.4s5.7㸪ᖹᆒ⏘ḟ㸸2.4 s0.1㸪ᖹᆒἪங㔞㸸34.9s0.7 kg/᪥㸪ᖹᆒయ㔜㸸646.7s6.3 kg㸪ᖹᆒ BCS㸸2.91s0.03㸧 㸪 㠀ඹᏑ⩌ࡣ n㸻109㸦ᖹᆒศፔᚋ᪥ᩘ㸸118.3s5.6㸪ᖹᆒ⏘ḟ㸸2.1s0.1㸪ᖹᆒἪங㔞㸸32.7 s0.7 kg/᪥㸪ᖹᆒయ㔜㸸626.6s6.5 kg㸪ᖹᆒ BCS㸸2.92s0.03㸧࡛࠶ࡾ㸪ᮍ⤒⏘∵࠾ࡅ ࡿඹᏑ⩌ࡣ n㸻55㸦ᖹᆒయ㔜㸸434.1s6.3 kg㸪ᖹᆒ᭶㱋㸸14.1s0.6㸧 㸪㠀ඹᏑ⩌࡛ࡣ n㸻 57㸦ᖹᆒయ㔜㸸446.9s8.0 kg㸪ᖹᆒ᭶㱋㸸14.2s0.5㸧࡛࠶ࡗࡓࠋ ⤒⏘∵࠾ࡼࡧᮍ⤒⏘∵ࢆྜࢃࡏ㸪ඹᏑ⩌㠀ඹᏑ⩌࡛ཷ⫾⋡ࢆẚ㍑ࡋࡓࡇࢁ㸪ඹᏑ⩌ ࡛᭷ពప࠸ཷ⫾⋡࡛࠶ࡗࡓ㹙ඹᏑ⩌ vs. 㠀ඹᏑ⩌㸻40.2㸣㸦74/184㸧 vs. 69.3㸣 㸦115/166㸧 㸹Fig. 4.4.㹛 ࠋࡲࡓ㸪⤒⏘∵ᮍ⤒⏘∵࡛ࡑࢀࡒࢀඹᏑ⩌㠀ඹᏑ⩌ࡢཷ⫾⋡ࢆ 83 ẚ㍑ࡋࡓࡇࢁ㸪⤒⏘∵ᮍ⤒⏘∵ࡶඹᏑ⩌࡛ప࠸ཷ⫾⋡࡛࠶ࡗࡓ㹙⤒⏘∵㸹ඹᏑ⩌ vs. 㠀ඹᏑ⩌㸻38.0㸣㸦49/129㸧 vs. 67.0㸣㸦73/109㸧 㸪ᮍ⤒⏘∵㸹ඹᏑ⩌ vs. 㠀ඹᏑ⩌㸻45.5㸣 㸦25/55㸧 vs. 73.7㸣㸦42/57㸧 㹛 ࠋࡲࡓ㸪ࣟࢪࢫࢸࢵࢡᅇᖐศᯒࡢ⤖ᯝࡽ㸪ᮏヨ㦂࡛ࡣ㸪 Ꮨ⠇㸪య㔜㸪ศፔᚋ᪥ᩘ㸪ங㔞㸪BCS࠾ࡼࡧ⏘ḟࡣཷ⫾⋡ᙳ㡪ࢆཬࡰࡉ࡞࠸ࡇࡀ᫂ࡽ ࡞ࡗࡓࠋ 4㸫5㸬⪃ᐹ ヨ㦂 1 ࠾࠸࡚⤒⏘Ἢங∵ࢆ⏝࠸࡚㸪W1DF 㯤యࡀྠ୍༸ᕢෆᏑᅾࡍࡿඹᏑ㛵ಀ ࡢሙྜ㸪ࡑࢀࡒࢀࡀᕥྑࡢ␗࡞ࡿ༸ᕢᏑᅾࡍࡿ㠀ඹᏑ㛵ಀࡢሙྜࡘ࠸࡚ W1DF 㯤యࡢⓎ⫱ືែ࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ࢆẚ㍑ࡋࡓࠋࡑࡢ⤖ᯝ㸪㯤యࡢⓎ⫱ືែ⾑୰ P4 ⃰ᗘືែᕪࡣࡳࡽࢀ࡞ࡗࡓࡀ㸪ඹᏑ⩌ࡢ W1DF ࡣࡁࡃⓎ⫱ࡍࡿࡇࡀ᫂ࡽ ࡞ࡗࡓࠋࡲࡓ㸪⤒⏘Ἢங∵࠾ࡼࡧᮍ⤒⏘∵ࢆ⏝࠸࡚㸪ேᕤᤵ⢭ᚋࡢཷ⫾⋡ࢆඹᏑ⩌㠀 ඹᏑ⩌࡛ẚ㍑ࡋࡓࡇࢁ㸪ඹᏑ⩌࡛పୗࡍࡿࡇࢆ᫂ࡽࡋࡓࠋ ➨ 3 ❶࡛ࡣ㸪㠀Ἢங∵ࢆ⏝࠸࡚ Day 6 ࠾ࡅࡿ W1DF ࡢᖹᆒ┤ᚄ⾑୰ P4 ⃰ᗘࡘ࠸ ࡚ඹᏑ⩌㠀ඹᏑ⩌࡛ẚ㍑ࡋࡓࡇࢁ㸪ඹᏑ⩌࡛ᖹᆒ┤ᚄࡣࡁࡃ㸪⾑୰ P4 ⃰ᗘࡣప࠸ W1DF ࠸࠺⤖ᯝࢆᚓࡽࢀࡓࠋ Ἢங∵ࢆ⏝࠸ࡓᮏヨ㦂࡛ࡣ⾑୰ P4 ⃰ᗘᕪࡣ࡞ࡗࡓࡀ㸪 ࡢᖹᆒ┤ᚄࡣඹᏑ⩌࡛ࡁࡃ࡞ࡾ㸪ᖹᆒ┤ᚄ㛵ࡋ࡚ࡣྠᵝࡢ⤖ᯝࡀᚓࡽࢀࡓࠋࡉࡽ㸪 㯤యࡢᖹᆒ┤ᚄᕪࡣ࡞ࡃ㸪➨ 3 ❶ࡢヨ㦂 2 ྠᵝࡢ⤖ᯝࡀᚓࡽࢀࡓࠋࡇࢀࡲ࡛㸪㧗Ἢ ங∵ࡢ⾑୰ P4 ⃰ᗘࡣᮍ⤒⏘∵ẚ࡚పࡃ࡞ࡿ㸦78㸧ࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿࡀ㸪୍᪉࡛㸪 㠀Ἢங∵Ἢங∵ࡢⓎ࿘ᮇ୰ࡢ⾑୰ P4 ⃰ᗘ᥎⛣ࢆẚ㍑ࡋࡓࡇࢁ㸪Ἢங∵࡛㯤యᮇ୰ ᮇ࠾࠸࡚㧗ࡃ᥎⛣ࡍࡿ㸦23㸧࠸࠺ሗ࿌ࡶ࠶ࡾ㸪ࡇࡢࡇࡣἪங∵ࡢங⏕⏘క࠺⫢ ⮚࡛ࡢࢫࢸࣟࢻ௦ㅰࡢ㐪࠸㸦100㸧ࡸ༸๓༸⬊ࡢࢧࢬࡀ␗࡞ࡿࡇ࡛ᙧᡂࡉࢀࡿ㯤 యࡢࢧࢬࡀࡁࡃ࡞ࡿࡇ㸦23㸪97㸧⏤᮶ࡍࡿ⪃࠼ࡽࢀ࡚࠸ࡿࠋᮏヨ㦂࡛ࡣඹᏑ ⩌㸪㠀ඹᏑ⩌ࡶᵝࠎ࡞ங㔞ࢆᣢࡗࡓἪங∵ࡀヨ㦂౪ヨࡉࢀࡓࡓࡵ㸪⾑୰ P4 ⃰ᗘ 84 㛵ࡋ࡚➨ 3 ❶ࡢヨ㦂 1 ␗࡞ࡿ⤖ᯝࡀᚓࡽࢀࡓࡢ࡛ࡣ࡞࠸᥎ ࡉࢀࡿࠋࡋࡋ㸪ୖ グ࡛㏙ࡓࡼ࠺ᮏヨ㦂࡛ࡣ㸪ඹᏑ⩌㠀ඹᏑ⩌㛫࡛⾑୰ P4 ⃰ᗘᕪࡣ࡞࠸ࡶ㛵ࢃࡽ ࡎ W1DF ࡢᖹᆒ┤ᚄࡀඹᏑ⩌࡛ࡁࡃⓎ⫱ࡋࡓࠋ ᮍ⤒⏘∵ࢆ⏝࠸ࡓヨ㦂࠾࠸࡚㸪W1DF ࡢࢧࢬࡣ㯤యྠ୍༸ᕢෆඹᏑࡍࡿሙྜඹᏑࡋ࡞࠸ሙྜ࡛ᕪࡣ࡞࠸࠸࠺ሗ࿌ 㸦40㸧ࡀᏑᅾࡍࡿࠋࡋࡋ㸪ඹᏑ⩌࡛ࡁࡃⓎ⫱ࡍࡿ㸦42㸧࠸࠺ሗ࿌ࡶᏑᅾࡋ㸪ᮏヨ 㦂࡛ࡣᚋ⪅ࡢ⤖ᯝ୍⮴ࡍࡿ⤖ᯝ࡞ࡗࡓࠋ➨ 3 ❶ࡢヨ㦂 2 ࡛ࡣ㸪Day 4 ࡲ࡛ࡢᖍ༸ ⬊ࡢᖹᆒ┤ᚄࡢ᥎⛣ඹᏑ⩌㠀ඹᏑ⩌࡛᭷ព࡞ᕪࡣ࡞࠸࠸࠺⤖ᯝࡀ♧ࡉࢀ㸪ᮏヨ㦂 1 ࡛ᚓࡽࢀࡓ⤖ᯝ▩┪ࡍࡿ⤖ᯝ࡛࠶ࡿࠋࡋࡋᮏヨ㦂࡛ࡢᖍ༸⬊ࡢᖹᆒ┤ᚄࡢ᥎⛣ ࢆࡳࡿ Day 4 ࡲ࡛ࡣ୧⩌㛫࡛ᕪࡀ࡞࠸ࡇࡽ㸪➨ 3 ❶ࡢヨ㦂 2 ࠾࠸࡚ࡶ Day 6 ௨ 㝆࠾࠸࡚ඹᏑ⩌࡛ࡁࡃⓎ⫱ࡋࡓྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋᐇ㸪➨ 3 ❶ࡢヨ㦂 1 ࡛ Day 8 ࠾࠸࡚ඹᏑ⩌࡛ࡢW1DF ࡢᖹᆒ┤ᚄࡀࡁࡗࡓࡇࡽࡶࡇࡢ௬ㄝࡣࡅࡽࢀࡿࠋ ᮏヨ㦂࡛ࡣ㸪ඹᏑ⩌㠀ඹᏑ⩌ࡢ㛫࡛⾑୰ P4 ⃰ᗘᕪࡣ࡞࠸ࡶ㛵ࢃࡽࡎ㸪W1DF ࡢ ᖹᆒ┤ᚄᕪࡀ⏕ࡌࡓࡇࡽ㸪W1DF ࡢ┤ᚄࡢᕪ␗ࡣ⾑୰ P4 ⃰ᗘ࡛ࡣ࡞ࡃ㸪㯤యࡢ ⨨㛵ಀ㉳ᅉࡍࡿࡇࡀ᥎ ࡉࢀࡿࠋ࣮࢝ࣛࢻࣉࣛ⏬ീデ᩿⨨ࢆ⏝࠸ࡓ㐣ཤࡢヨ㦂 ࡛ࡣ㸪㯤య W1DF ࡀྠ୍༸ᕢෆ࡛Ⓨ⫱ࡋ࡚࠸ࡿሙྜ㸪༸ᕢࡢ⾑⟶ᢠ್ࡀపࡃ㸪༸⬊ ቨ⾑ὶࡀࡁ࠸ࡇࢆሗ࿌ࡋ࡚࠸ࡿ㸦42㸧 ࠋ༸⬊ቨ⾑ὶࡣ㸪㑅ᢤࡉࢀࡿ༸⬊ࡸ㛢㙐ࡋ࡚࠸ ࡃ༸⬊ࢆホ౯ࡍࡿᣦᶆࡋ࡚⏝࠸ࡽࢀ࡚࠾ࡾ㸦2㸪42㸧 㸪ࡲࡓ㸪༸๓༸⬊ࡢ༸⬊ቨ⾑ὶ 㸪ࡑࡢᚋࡢཷ⫾ᛶ㸦84㸧ࡸయእཷ⢭ᚋࡢ⬇ࡢⓎ⫱⋡㸦86㸧ࡣṇࡢ㛵ಀࡀᏑᅾࡍࡿࡇ ࡽ㸪༸⬊ቨ⾑ὶࡣ༸⬊ࡢάຊࢆホ౯ࡍࡿᣦᶆࡉࢀ࡚࠸ࡿࠋᮏヨ㦂࠾࠸࡚ࡶ㸪ඹ Ꮡ⩌ࡢ W1DF ࡢ༸⬊ቨ⾑ὶࡀࡁࡗࡓࡇࡀ᥎ ࡉࢀ㸪ඹᏑ⩌ࡢ W1DF 㓟⣲㸪ᰤ㣴 ࡉࡽࡣ࣍ࣝࣔࣥࡀࡼࡾከࡃ౪⤥ࡉࢀࡿ㸦109㸧ࡇ࡛㸪Ⓨ⫱ࡀࡁࡃ࡞ࡗࡓ᥎ᐹࡉࢀ ࡿࠋࡇࡢࡇࡣ㸪W1DF ࡢⓎ⫱ࡸᛶ㉁ཬࡰࡍせᅉࡋ࡚㸪㯤యྠ୍༸ᕢෆ࡛Ⓨ⫱ࡍ ࡿྰ࠸࠺⨨ⓗ≧ἣࡢᙳ㡪ࡀᏑᅾࡍࡿࡇࢆ♧၀ࡋ࡚࠸ࡿࠋ 㐣ཤࡢሗ࿌࠾࠸࡚㸪W1DF ࡢⓎ⫱ືែࡸࡑࢀక࠺࣍ࣝࣔࣥືែࡣヲ⣽ㄪࡽࢀ 85 ࡚ࡁࡓࠋࡋࡋ࡞ࡀࡽ㸪W1DF ࡀ⦾Ṫ⏕⌮㸪≉ཷ⫾ᛶཬࡰࡍᙳ㡪㛵ࡋ࡚ࡣ༑ศ ㄪᰝࡉࢀ࡚ࡇ࡞ࡗࡓࠋ➨ 3 ❶ࡢヨ㦂 1㸪2 ࠾ࡼࡧᮏ❶ࡢヨ㦂 1 ࡽ㸪W1DF ࡢᛶ㉁ࡣ㯤 యྠ୍༸ᕢෆ࡛ඹᏑࡍࡿྰ࠸࠺ࡇ࡛␗࡞ࡿࡇࡀ᥎ ࡉࢀࡿࠋࡇࢀࡲ࡛㸪㯤 యࡀᏑᅾࡍࡿ༸ᕢྠഃࡢᏊᐑゅ࠾ࡅࡿᏊᐑෆ⭷࡛ࡣ㸪ᑐഃࡢᏊᐑゅ࠾ࡅࡿᏊᐑ ࠋࡲࡓ㸪༸๓༸⬊ࡀᏑᅾ ෆ⭷ẚ⤌⧊୰ P4 ⃰ᗘࡀ㧗࠸ࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸦20㸧 ࡍࡿ༸ᕢྠഃࡢ༸⟶ෆ E2 ⃰ᗘࡣ㸪ᑐഃࡢࡶࡢẚ㍑ࡋ࡚㧗࠸ࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ 㸦99㸧 ࠋ࢘ࢩ࡛ࡣ㸪㯤యࡀᙧᡂࡉࢀ࡚࠸ࡿഃྠഃࡢᏊᐑゅ࡛╔ᗋ㸪ዷፎࡋࡸࡍ࠸ࡇࡀ ▱ࡽࢀ࡚࠸ࡿ㸦91㸧 ࠋࡇࢀࡽࡢᐇࡽ㸪༸ᕢྠഃࡢᏊᐑゅࡣᐦ᥋࡞ᒁᡤࡢࡘ࡞ࡀࡾ ࡀᏑᅾࡍࡿࡇࡀ♧၀ࡉࢀࡿࠋ ࣈࢱࢆ⏝࠸ࡓሗ࿌࠾࠸࡚㸪 ༸⟶ྥ࠺⾑⟶ࡢ⾑୰ P4㸪 A4 ࠾ࡼࡧ E2 ⃰ᗘࡣᮎᲈ⾑୰ࡢ⃰ᗘẚ㍑ࡋ࡚㧗࠸ࡇࡽ㸪 ༸ᕢ㟼⬦ࡽᏊᐑࡸ༸⟶ ⾑ᾮࢆ౪⤥ࡍࡿ⾑⟶㐃⤖ࡋ࡚࠸ࡿ༸ᕢື⬦ᛶࢫࢸࣟࢻ࣍ࣝࣔࣥࡀᑐྥὶࡼ ࡾ⛣⾜ࡋ࡚࠸ࡿࡇࡀ᥎ ࡉࢀ࡚࠸ࡿ㸦47㸧 ࠋヨ㦂 2 ࠾࠸࡚㸪ேᕤᤵ⢭ᚋࡢཷ⫾⋡ࡣ⤒ ⏘∵ᮍ⤒⏘∵ඹඹᏑ⩌࡛పୗࡍࡿࡇࢆ♧ࡋࡓࠋࡲࡓ㸪ங㔞㸪⏘ḟ㸪ศፔᚋ᪥ᩘ㸪 య㔜㛵ࢃࡽࡎ㸪ඹᏑ⩌࡛ཷ⫾ᛶࡀప࠸ࡇࡽ㸪ཷ⫾ᛶࢆỴᐃࡍࡿᅉᏊࡋ࡚ W1DF 㯤య࡛ࡢ⨨㛵ಀࡀࡁࡃᙳ㡪ࢆ࠼࡚࠸ࡿྍ⬟ᛶࡀ⪃࠼ࡽࢀࡿࠋⓎ࿘ᮇ୰ࡢ⾑୰ E2 ⃰ᗘࢆ ᐃࡋࡓࡇࢁ㸪➨ 1 ༸⬊ἼࡀⓎ⫱ࡍࡿᮇ㛫⾑୰ E2 ⃰ᗘࡀୖ᪼ࡍࡿࡇࡀሗ ࿌ࡉࢀ࡚࠸ࡿࡇࡽ㸪W1DF ࡣ⾑୰ E2 ⃰ᗘࢆୖ᪼ࡉࡏࡿ༑ศ࡞ E2 ࢆ⏘⏕ࡋ࡚࠸ࡿ ࡇࡀ࠺ࡀ࠼ࡿ㸦79㸧 ࠋࡑࡢࡓࡵ㸪W1DF ࡀᏑᅾࡍࡿ༸ᕢྠഃࡢ༸⟶ࡸᏊᐑゅ࡛ࡣ E2 ⃰ᗘࡀ㧗ࡃ࡞ࡿ᥎ ࡉࢀࡿࠋ࢘ࢩࡢ༸⟶ୖ⓶⣽⬊ࢆ⏝࠸ࡓ in vitro ࡢᐇ㦂࠾࠸࡚㸪 E2 ࡣ࢚ࢫࢺࣟࢤࣥཷᐜయȘ P4 ཷᐜయࡢ mRNA Ⓨ⌧㔞ࢆୖ᪼ࡉࡏࡿ࠸࠺ሗ࿌ࡀ࠶ࡿ 㸦96㸧 ࠋࡲࡓ㸪࢘ࢩࡢᏊᐑෆ⭷⣽⬊ࢆ⏝࠸ࡓ in vitro ࡢᐇ㦂࠾࠸࡚㸪E2 ࡣ࢚ࢫࢺࣟࢤࣥ ཷᐜయ࠾ࡼࡧ P4 ཷᐜయⓎ⌧ࢆୖ᪼ࡉࡏࡿ㸦102㸧 ࠋࡇࢀࡽࡢࡇࡽ㸪E2 ࡣ༸⟶ࡸᏊᐑෆ ⭷ࡢࢫࢸࣟࢻ࣍ࣝࣔࣥཷᐜయࡢⓎ⌧ࢆไᚚࡍࡿࡇࡀ♧၀ࡉࢀࡿࠋW1DF ࡀ㯤యඹ Ꮡࡍࡿሙྜ㸪W1DF ࡽ⏘⏕㸪ᨺฟࡉࢀࡿ E2 ࡀཷ⢭༸ࡸ⬇ࡀᏑᅾࡍࡿྠഃࡢ༸⟶ࡸᏊᐑ 86 ゅᑐࡋ࡚ᒁᡤⓗస⏝ࡋዷፎᡂ❧㛵ࢃࡿᶵ⬟ᙳ㡪ࢆ࠼ࡿࡇ࡛㸪ዷፎᡂ❧ᑐ ࡋ࡚㈇ࡢᙳ㡪ࢆ࠼࡚࠸ࡿࡇࡀ᥎ ࡉࢀࡿࠋࡲࡓ㸪ྠ୍༸ᕢෆᏑᅾࡍࡿ W1DF ࡀ㯤 యᑐࡋ࡚㸪㯤యᙧᡂࡸ P4 ⏘⏕⬟ࢆᢚไࡍࡿ㸪ࡲࡓࡣ㯤యࡢᏑ⥆ᮇ㛫ࢆ▷ࡃࡍࡿࡇ࡛ ཷ⫾ᛶࡀపୗࡋ࡚࠸ࡿྍ⬟ᛶࡶ⪃࠼ࡽࢀࡿࠋࡋࡋ㸪ᮏ❶ࡢヨ㦂 2 ࡽ㯤యⓎ⫱ືែࡸ ⾑୰ P4 ⃰ᗘඹᏑ⩌㠀ඹᏑ⩌࡛ᕪࡀࡳࡽࢀ࡞ࡗࡓࡇࡽ㸪 ඹᏑࡍࡿ W1DF ࡼࡿ 㯤యࡢᢚไస⏝ࡢྍ⬟ᛶࡣప࠸⪃࠼ࡽࢀࡿࠋ୍᪉㸪㠀Ἢங⤒⏘∵ࢆ⏝࠸ࡓ➨ 3 ❶ࡢ ヨ㦂 1 ࡛ࡣඹᏑ⩌࡛⾑୰ P4 ⃰ᗘࡀపࡗࡓࡇࡸ㸪ᮏヨ㦂࠾࠸࡚ࡶⓎ࿘ᮇయࢆ㏻ ࡋ࡚ࡢ㯤యࡢᏑ⥆ᮇ㛫ࢆㄪᰝࡋ࡚࠸࡞࠸ࡓࡵ㸪ᚋ᭦࡞ࡿ◊✲ࢆ⾜࠺ᚲせࡀ࠶ࡿ⪃࠼ ࡿࠋ ᮏ❶࡛ࡣ㸪 W1DF ࡸ㯤యࡢⓎ⫱ືែ࠾ࡼࡧ⾑୰ P4 ⃰ᗘࢆẚ㍑ࡍࡿࡇ࡛㸪 㯤య W1DF ࡀྠ୍༸ᕢෆඹᏑࡲࡓࡣඹᏑࡋ࡞࠸࠸࠺༸ᕢෆ࡛ࡢ⨨㛵ಀࡀ⤒⏘Ἢங∵࠾ࡅ ࡿ W1DF ࡢⓎ⫱㛵ࢃࡿせᅉࡋ࡚㔜せ࡛࠶ࡿࡇࢆ♧ࡋࡓࠋࡉࡽ㸪ࡑࡢ⨨㛵ಀࡀ ཷ⫾ᛶࡲ࡛ࡁࡃᙳ㡪ࢆཬࡰࡍࡇࢆ᫂ࡽࡋ㸪W1DF ࡢᛶ㉁࠾ࡼࡧ⦾Ṫ⏕⌮ཬ ࡰࡍᙳ㡪ࢆ♧ࡋࡓࠋ 4㸫6㸬ᑠᣓ ᮏ❶࡛ࡣ㸪⤒⏘Ἢங∵ࢆ⏝࠸࡚㸪W1DF ࡀ㯤యྠ୍༸ᕢෆ࡛ඹᏑࡋ࡚Ⓨ⫱ࡍࡿሙྜ㸪 W1DF ࡢᖹᆒ┤ᚄ᥎⛣ࡀ㠀ඹᏑẚ࡚ࡁࡃⓎ⫱ࡍࡿࡀ㸪㯤యࡢⓎ⫱ືែࡸ⾑୰ P4 ⃰ ᗘࡢ᥎⛣㐪࠸ࡣࡳࡽࢀ࡞࠸ࡇࢆ♧ࡋࡓࠋࡲࡓ㸪ேᕤᤵ⢭ᚋ࠾ࡅࡿ W1DF ࡀ㯤య ඹᏑࡍࡿሙྜ㠀ඹᏑࡢሙྜ࡛ࡢࡑࡢᚋࡢཷ⫾⋡ࢆẚ㍑ࡋࡓࡇࢁ㸪ඹᏑ⩌࡛ཷ⫾⋡ࡀ ప࠸ࡇࢆ♧ࡋ㸪W1DF ࡢ⦾Ṫ⏕⌮࠼ࡿᙳ㡪ࢆ᫂ࡽࡋࡓࠋ 87 Cross-sectional area of corpus luteum (mm2) 700 600 IG CG 500 400 G : NS D : P < 0.001 300 G,D : NS 200 100 Day2 Day4 Day6 Day8 Day10 Days from follicular wave emergence Fig. 4.1. Comparative changes of corpus luteum (CL) of IG and CG on Day 2, 4, 6, 8, 10. Day 0 = The day of the first follicular wave emergence. Effect of group (G) (NS), day (D) (P < 0.001) and interaction between group and day (G,D) (NS). IG; Ipsilateral group: Dominant follicle (DF) ipsilateral to the CL. CG; Contralateral group: DF contralateral to the CL. Values are mean r SEM of each point. 88 Diameter of dominant follicle (mm) 20 18 IG CG 16 14 12 G : P < 0.01 10 D : P < 0.001 G,D : P < 0.01 8 6 Day2 Day4 Day6 Day8 Day10 Days from follicular wave emergence Fig. 4.2. Comparative changes of the first follicular wave dominant follicle (DF) of IG and CG on Day 2, 4, 6, 8, 10. Day 0 = The day of the first follicular wave emergence. Effect of group (G) (P < 0.01), day (D) (P < 0.001) and interaction between group and day (G,D) (P < 0.01). IG; ipsilateral group: DF ipsilateral to the corpus luteum (CL). CG; contralateral group: DF contralateral to the CL. The asterisk denotes difference between each point, : P < 0.05. Values are mean r SEM of each point. 89 Plasma P concentration of P4 (ng/ml) Plasma (ng/ml) 4 concentration 7.0 IG CG 6.0 5.0 4.0 3.0 G : NS 2.0 D : P < 0.001 1.0 G,D : NS 0 Day2 Day4 Day6 Day8 Day10 Daysfrom fromfollicular follicular wave wave emergence Days emergence Fig. 4.3. Comparative changes of plasma P4 concentration of IG and CG on Day 2, 4, 6, 8, 10. Day 0 = The day of the first follicular wave emergence. Effect of group (G) (NS), day (D) (P < 0.001) and interaction between group and day (G,D) (NS). IG; ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG; contralateral group: DF contralateral to the CL. Values are mean r SEM of each point. 90 IG 80 73.7% (42/57) 67.0% (73/109) 70 Conception rate (%) CG 60 50 45.5% (25/55) 38.0% (49/129) 40 30 20 10 0 Lactating dairy cows Dairy heifers Figure 4.4. Conception rates in IG and CG. Conception rates were different between IG and CG in both lactating dairy cows and dairy heifers. IG; ipsilateral group: Dominant follicle (DF) ipsilateral to the corpus luteum (CL). CG; contralateral group: DF contralateral to the CL. The asterisk denotes difference between each point, : P < 0.01. 91 ➨㸳❶ ⥲ྜ⪃ᐹ 92 ㏆ᖺࡢ␆⏘⌧ሙ࠾ࡅࡿከ㢌࠾ࡼࡧங∵࠾ࡅࡿ㧗Ἢஙక࠸㸪ⓎⓎぢ⋡ࡢప ୗ࠾ࡼࡧཷ⫾⋡పୗࡀၥ㢟࡞ࡗ࡚࠸ࡿࠋࡇࡢၥ㢟ᑐฎࡍࡿࡓࡵ㸪ᐃேᕤᤵ⢭ࣉ ࣟࢢ࣒ࣛࡢ㛤Ⓨࢆ୰ᚰࡍࡿ◊✲ࡀ┒ࢇ⾜ࢃࢀ࡚࠸ࡿࡀ㸪⦾Ṫᡂ⦼ࡢྥୖࡣ༑ศ㐩 ᡂ࡛ࡁ࡚࠸࡞࠸ࠋࡇࡢࡇࡣ㸪࢘ࢩࡢཷ⫾㛵ࢃࡿ⦾Ṫ⏕⌮ᑐࡍࡿ⌮ゎࡀᮍࡔ༑ศ ࡛࠶ࡿࡇࢆ♧ࡋ࡚࠸ࡿࠋࡑࡇ࡛㸪⚾ࡣ㸪ᮍࡔ᫂ࡽࡉࢀ࡚࠸࡞࠸࢘ࢩࡢ⦾Ṫ⏕⌮ࢆ ᫂ࡽࡍࡿࡇࢆ┠ⓗ࢘ࢩࡢⓎ࿘ᮇ୰Ⓨ⫱ࡍࡿ༸⬊Ἴ㸪≉㸪ࢇࡀ༸ ࡍࡿࡇ࡞ࡃ㛢㙐ࡋ࡚ࡋࡲ࠺➨ 1 ༸⬊Ἴࡢᖍ༸⬊╔┠ࡋࡓࠋ➨ 1 ༸⬊ἼࡣⓎ⤊ ᚋࡢ༸┤ᚋࡽ༸ᕢෆ࡛」ᩘࡢᑠ༸⬊ࡢⓎ⫱ࡀ㛤ጞࡉࢀࡿࡇ࡛Ⓨ⌧ࡋ㸪Ⓨ⫱㛤ጞ ࡽ 2㹼3 ᪥ᚋࡑࡢ୰ࡽ᭱ࡢ༸⬊ࡢࡳࡀ㑅ᢤࡉࢀ㸪ᖍ༸⬊࡞ࡾ㸪ࡑࢀ௨እࡢ༸⬊ ࡣ࡚㛢㙐ࡋ࡚ࡋࡲ࠺ࠋࡇࡢ➨ 1 ༸⬊Ἴᖍ༸⬊㸦W1DF㸧ࡘ࠸࡚㸪༸⬊ࡢⓎ⫱ືែ ࡸ㑅ᢤࡢ࣓࢝ࢽࢬ࣒ࢆ᫂ࡽࡍࡿࡓࡵ㸪࣍ࣝࣔࣥືែࢆྵࡵ࡚ࡇࢀࡲ࡛ከࡃࡢ◊✲ ࡀ⾜ࢃࢀ࡚ࡁࡓࡀ㸪W1DF ⮬యࡢᛶ㉁ࡢ≉ᚩࡸ⦾Ṫ⏕⌮࠾ࡅࡿព⩏ࡘ࠸࡚ࡣࢇ ◊✲ࡀ⾜ࢃࢀ࡚ࡇ࡞ࡗࡓࠋࡑࡇ࡛㸪ࡇࡢ W1DF ࡢᛶ㉁㸪Ⓨ⫱㛵ࢃࡿせᅉ㸪ࡉࡽ㸪 ཷ⫾࠼ࡿᙳ㡪ࢆ᫂ࡽࡍࡿࡓࡵ㸪ᖍ༸⬊ࡢⓎ⫱ືែ㸪༸⬊ቨ⾑ὶࡢ᥎⛣㸪 ༸┤๓ࡢ⾑୰ E2 ⃰ᗘ㸪༸๓ᚋ࠾ࡅࡿ༸⬊ᾮ୰ࡢ P4㸪E2 ࠾ࡼࡧ A4 ⃰ᗘ㸪༸ᚋ࠾ ࡅࡿ㯤యࡢⓎ⫱ືែ࠾ࡼࡧ⾑୰ P4 ⃰ᗘࡢ᥎⛣㸪㯤యࡢ⨨㛵ಀ㸪ࡑࡋ࡚ཷ⫾ᛶࢆホ౯ ࡋࡓࠋ ➨ 2 ❶࡛ࡣ㸪⤒⏘㠀Ἢங∵ࢆ⏝࠸࡚ W1DF ࡢ≉᭷ࡢᛶ㉁ࢆ᫂ࡽࡍࡿࡓࡵ㸪➨ 2 ༸ ⬊Ἴᖍ༸⬊㸦W2DF㸧ࡢẚ㍑ࢆ⾜ࡗࡓ㸦W1DF㸸n㸻7㸪W2DF㸸n㸻7㸧 ࠋヨ㦂 1 ࡛ࡣ㸪 Day 3 ࡽ 6㸦Day 0㸻༸⬊ἼⓎ⌧᪥㸧ࡲ࡛ࡢᖍ༸⬊ࡢⓎ⫱ືែ⾑୰ P4 ⃰ᗘࡢ᥎⛣㸪 ࡑࡋ࡚ Day 6 ࠾ࡅࡿ༸⬊ቨ⾑ὶࢆ࣮࢝ࣛࢻࣉࣛ⏬ീデ᩿⨨ࢆ⏝࠸࡚ ᐃࡋࡓࠋࡑࡢ ⤖ᯝ㸪W1DF ࡣ W2DF ẚ㍑ࡋ࡚ᖹᆒ┤ᚄࡣࡁࡃ᥎⛣ࡋ㸪Day 6 ࡢᖹᆒ┤ᚄࡣ᭷ព ࡁࡗࡓࠋᖍ༸⬊࡞ࡗ࡚ࡽࡢⓎ⫱⋡ࡀ W1DF ࡛ࡁ࠸ࡇࡀ㸪ᖹᆒ┤ᚄࡢᕪ␗ ࢆࡶࡓࡽࡋࡓཎᅉ࡛࠶ࡿ⪃࠼ࡽࢀࡓࠋࡲࡓ㸪⾑୰ P4 ⃰ᗘ᥎⛣ࢆ୧⩌㛫࡛ẚ㍑ࡋࡓࡇ 93 ࢁ㸪W1DF ࡀⓎ⫱ࡍࡿᮇ㛫ࡢ⾑୰ P4 ⃰ᗘࡣ㸪W2DF ࡀⓎ⫱ࡍࡿᮇ㛫ࡢ⾑୰ P4 ⃰ᗘࡼࡾ పࡃ᥎⛣ࡍࡿࡇࡀ♧ࡉࢀࡓࠋᮍ⤒⏘∵ࢆ⏝࠸ࡓヨ㦂࠾࠸࡚㸪W1DF ࡀⓎ⫱ࡍࡿᮇ ⭐ෆ␃⨨ᆺ P4 〇ࢆᤄධࡋࡓࡇࢁ㸪ᤄධࡋ࡞ࡗࡓሙྜẚ㍑ࡋ࡚ᖍ༸⬊ࡢ᭱ ┤ᚄࡀᑠࡉࡃ࡞ࡾ㸪⾑୰ P4 ⃰ᗘࡀ㧗࠸᮲௳ୗ࡛ࡣᖍ༸⬊ࡢⓎ⫱ࡀᢚไࡉࢀࡿࡇࡀ♧ ࡉࢀ࡚࠸ࡿࠋᖍ༸⬊㑅ᢤࡉࢀ㸪Ⓨ⫱ࡀ⥔ᣢࡉࢀࡿࡓࡵࡣ㢛⢏ᒙ⣽⬊࡛ LHr ࡢⓎ⌧ ࡀᚲ㡲࡛࠶ࡾ㸪ᖍ༸⬊ࡣ LH ౫ᏑᛶⓎ⫱ࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋ⾑୰ P4 ⃰ᗘࡀప ࠸㸪⬻ୗᆶయ๓ⴥࡽศἪࡉࢀࡿ LH ࡢࣃࣝࢫ㢖ᗘࡀ㧗ࡃ࡞ࡿࡇࡀ▱ࡽࢀ࡚࠾ࡾ㸪 ᮏ◊✲࠾࠸࡚ W1DF ࡢⓎ⫱ࡀࡁ࠸ࡢࡣ㧗࠸ LH ࣃࣝࢫ㢖ᗘ㉳ᅉࡍࡿ᥎ ࡉࢀࡿࠋ ࡲࡓ㸪ᮏ◊✲࡛ࡣ༸⬊ቨ⾑ὶ࠾࠸࡚㸪W1DF ࡛ࡁ࠸ࡇࢆ♧ࡋࡓࠋᖍ༸⬊ࡀ㑅ᢤ ࡉࢀࡿ๓ᚋ࡛༸⬊ቨ⾑ὶࡀቑຍࡍࡿࡇ㸪㛢㙐ࡍࡿ༸⬊࡛ࡣ༸⬊ቨ⾑ὶࡀᾘኻࡍࡿࡇ ࡽ㸪༸⬊ቨ⾑ὶࡣ༸⬊ࡢάຊࡢᣦᶆ࡞ࡿࡓࡵ㸪W1DF ࡢάຊࡣ W2DF ẚ㍑ࡋ࡚㧗 ࠸ࡇࡀ᥎ ࡉࢀࡓࠋࡑࡇ࡛㸪ヨ㦂 2 ࡛ࡣ㸪PGF2α ࢆ Day 6㸪GnRH ࢆ Day 8 ᢞࡋ㸪 Ⓨ࠾ࡼࡧ༸ㄏ㉳ࢆ⾜࡞࠸㸪ࡑࢀక࠺ W1DF W2DF ࡢᛶ㉁ࢆẚ㍑ࡋࡓࠋGnRH ᢞ 㸦Day 8㸧ࡢ⾑୰ E2 ⃰ᗘࢆẚ㍑ࡋࡓࡇࢁ㸪W1DF ࡛᭷ព㧗ࡗࡓࠋᖹᆒ┤ᚄࡣ W1DF ࡛ࡁࡃ᥎⛣ࡋࡓࠋࡲࡓ㸪༸⬊ቨ⾑ὶࡢ᥎⛣࠾࠸࡚ࡣ㸪GnRH ᢞᚋࡢ༸⬊ቨ ⾑ὶࡣ W1DF ࡛᭷ព㧗ࡗࡓࠋࡇࢀࡣ GnRH ᢞ๓ᚋ࡛ࡢ⾑ὶቑຍࡀ W1DF ࡛ࡁ ࡗࡓࡇࡀཎᅉ࡛࠶ࡿ⪃࠼ࡽࢀࡿࠋᐃேᕤᤵ⢭ἲࡢ 1 ࡘ࡛࠶ࡿ࢜ࣈࢩࣥࢡࢆ⏝࠸ ࡓヨ㦂࠾࠸࡚㸪GnRH ᢞࡢ⾑୰ E2 ⃰ᗘࡑࡢᚋࡢཷ⫾ྍ⬟ᛶṇࡢ┦㛵㛵ಀࡀࡳ ࡽࢀ࡚࠸ࡿࠋࡉࡽ㸪GnRH ᢞ 26 㛫ᚋࡢ༸⬊ቨ⾑ὶேᕤᤵ⢭ᚋࡢཷ⫾ࡢ᭷↓࠾ࡼ ࡧࡑࡢ༸๓༸⬊ࡽᚓࡽࢀࡿ༸Ꮚࡢయእཷ⢭ᚋࡢ⬇Ⓨ⫱ࢆẚ㍑ࡋࡓ◊✲࡛ࡣ㸪ཷ⫾ ∵ẚ㍑ࡋ࡚ཷ⫾∵ࡢ༸⬊ቨ⾑ὶࡀ㧗ࡃ㸪ࡉࡽ⬇┙⬊ࡲ࡛Ⓨ⫱ࡋࡓ༸Ꮚࢆ᭷ࡋࡓ༸ ⬊࡛༸⬊ቨ⾑ὶࡀ㧗ࡗࡓࠋࡋࡓࡀࡗ࡚㸪W1DF ࡣཷ⫾ᛶࡢ㧗࠸άຊࡢ࠶ࡿ༸⬊࡛࠶ࡿ ᥎ ࡉࢀࡿࠋヨ㦂 3 ࡛ࡣ㸪W1DF ࡢᛶ㉁ࢆࡉࡽヲ⣽ホ౯ࡍࡿࡓࡵ㸪GnRH ᢞ ๓㸦W1DF㸸n㸻7㸪W2DF㸸n㸻7㸧ᢞ 24 㛫ᚋ㸦W1DF㸸n㸻6㸪W2DF㸸n㸻6㸧ࡢ༸ 94 ⬊ᾮ୰ࡢᛶࢫࢸࣟࢻ࣍ࣝࣔࣥ㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡍࡿ mRNA Ⓨ⌧㔞ࢆẚ㍑ࡋࡓࠋ GnRH ᢞ๓ࡢ༸⬊ࡘ࠸࡚ࡣ㸪ヨ㦂 2 ࠾ࡅࡿ⾑୰ E2 ⃰ᗘࡢᕪ␗㛵ࢃࡿ༸⬊ᶵ⬟ࢆ ホ౯ࡍࡿࡓࡵ㸪༸⬊ᾮ E2㸪A4 ࠾ࡼࡧ㢛⢏ᒙ⣽⬊࡛ࡢ CYP19A1 mRNA Ⓨ⌧㔞ࢆẚ㍑ࡋ ࡓࠋࡉࡽ㸪W1DF ࡳࡽࢀࡓ GnRH ᢞ 24 㛫ᚋࡢ༸⬊ቨ⾑ὶࡢ㧗್ࡀ㸪LH ࢧ࣮ࢪ ᑐࡍࡿᛂᛶࡢᕪ␗㉳ᅉࡍࡿࡢࢆㄪࡿࡓࡵ㸪㢛⢏ᒙ⣽⬊࠾ࡅࡿ LHr mRNA Ⓨ ⌧㔞ࢆẚ㍑ࡋࡓࠋࡑࡢ⤖ᯝ㸪༸⬊ᾮ E2 ࠾ࡼࡧ A4 ⃰ᗘࡣ W1DF ࡛㧗࠸್ࢆ♧ࡋࡓࡀ㸪 ࣐ࣟࢱ࣮ࢮ㸦CYP19A1㸧ࡢ mRNA Ⓨ⌧㔞᭷ព࡞ᕪࡣ࡞ࡗࡓࠋA4 ࡣෆ༸⬊⭷⣽⬊࡛ ⏘⏕ࡉࢀ㸪㢛⢏ᒙ⣽⬊࡛࣐ࣟࢱ࣮ࢮࡼࡾ E2 ኚࡉࢀࡿࠋ࣐ࣟࢱ࣮ࢮάᛶᕪ ␗ࡀ࡞ࡗࡓࡇࡽ㸪 E2 ࡢᇶ㉁࡛࠶ࡿ A4 ࢆ⏘⏕ࡍࡿෆ༸⬊⭷⣽⬊ࡢᶵ⬟ࡀ㧗࠸ࡇࡀ㸪 ᮏ◊✲࡛ࡢW1DF࠾ࡅࡿGnRHᢞࡢ㧗࠸⾑୰E2 ⃰ᗘࢆࡶࡓࡽࡋࡓ᥎ ࡉࢀࡓࠋ ࡲࡓ㸪LHr mRNA Ⓨ⌧㔞ࡣ W1DF ࡛᭷ព㧗ࡗࡓࠋ࢘ࢩࡢ㢛⢏ᒙ⣽⬊࠾࠸࡚㸪LH ࡣ LHr mRNA Ⓨ⌧㔞ࢆቑຍࡉࡏࡿሗ࿌ࡉࢀ࡚࠸ࡿࠋW1DF ࡣప࠸⾑୰ P4 ⃰ᗘ᮲௳ୗ ࡛Ⓨ⫱ࡋ࡚࠸ࡿࡓࡵ㸪ẚ㍑ⓗ㧗࠸㢖ᗘࡢ LH ࣃࣝࢫࡉࡽࡉࢀ࡚࠸ࡿࡇࡀணࡉࢀ㸪 ࡇࡢࡇࡀ㢛⢏ᒙ⣽⬊࡛ࡢ㧗࠸ LHr mRNA Ⓨ⌧㔞ࢆࡶࡓࡽࡋࡓ⪃࠼ࡽࢀࡿࠋḟ㸪 GnRH ᢞ 24 㛫ᚋࡢ㢛⢏ᒙ⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿ mRNA ࢆẚ㍑ࡋࡓࠋLH ࢧ࣮ࢪ᭚㟢ᚋ ࡢ༸⬊ෆ࡛ࡣ㸪㢛⢏ᒙ⣽⬊࠾ࡼࡧ༸⬊⭷⣽⬊ࡣ㯤య⣽⬊ᛴ㏿ኚࡋ㸪E2 ⏘⏕ࡽ P4 ⏘⏕ᶵ⬟ࡀࢩࣇࢺࡋ࡚࠸ࡃࠋຍ࠼࡚㸪㯤యᙧᡂࡣ⾑⟶ᙧᡂࡀ㔜せ࡛࠶ࡾ㸪VEGF ࡸ FGF ࡣ⾑⟶ᙧᡂࡢ୰ᚰⓗ࡞ᙺࢆᢸࡗ࡚࠸ࡿࠋ ༸๓༸⬊ࡸ㯤యᙧᡂ㸪ࡇࢀࡽ ࡢ⾑⟶ᙧᡂ㛵ࢃࡿᅉᏊࡢᶵ⬟ࢆ㜼ᐖࡍࡿࡑࡢᚋᙧᡂࡉࢀࡿ㯤యࡢࢧࢬࡸᶵ⬟ࡀ పୗࡍࡿࡇࡀ▱ࡽࢀ࡚࠸ࡿࠋࡉࡽ㸪༸⬊ෆ࡛ࡣ LH ࢧ࣮ࢪ᭚㟢ᚋ㸪PG 㢮㸦PGF2α㸪 PGE2 ➼㸧ࡢ⃰ᗘࡀୖ᪼ࡍࡿࡀ㸪ࡇࡢ PGE2 ྜᡂࢆ㜼ᐖࡍࡿ༸⬊◚ࡀ㜼ᐖࡉࢀࡿࡇ ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋ ࡲࡓ㸪 ⾑⟶ᣑᙇᅉᏊ࡛࠶ࡿ NO ࡢྜᡂ㓝⣲࡛࠶ࡿ iNOS ࠾ࡼࡧ eNOS ࡣ༸⬊ෆ࡛Ⓨ⌧ࡋ࡚࠸ࡿࠋࡇࢀࡽࡢᅉᏊࢆẚ㍑ࡍࡿࡇ࡛㸪LH ࢧ࣮ࢪᑐࡍࡿᛂᛶ ࡸ W1DF ࡛ࡢ༸⬊ቨ⾑ὶࡢ㧗್ࡢཎᅉ㸪ࡉࡽࡣࡑࡢᚋᙧᡂࡉࢀࡿ㯤యࡢᶵ⬟࠼ 95 ࡿᙳ㡪ࡘ࠸࡚ホ౯࡛ࡁࡿ⪃࠼ࡓࠋCOX-2㸪PGES ࠾ࡼࡧ PGFS mRNA Ⓨ⌧㔞᭷ព ࡞ᕪࡣ࡞ࡗࡓࠋ ࡇࡢࡇࡣ㸪༸㐣⛬ᚲ㡲࡞ࡇࢀࡽࡢᅉᏊࡢⓎ⌧㔞ཬࡰࡍ㸪LH ࢧ࣮ࢪᑐࡍࡿᛂᛶW1DFW2DFࡢ㛫࡛ᕪ␗ࡣ࡞࠸ࡇࢆ♧ࡋ࡚࠸ࡿ⪃࠼ࡽ ࢀ㸪ࡇࢀࡽࡢᅉᏊࡢⓎ⌧㔞ࡢ㐪࠸ࡀ༸⬊ቨ⾑ὶࡢᕪ␗ࢆࡶࡓࡽࡋࡓཎᅉ࡛࠶ࡿࡣ⪃࠼ ࡽࢀ࡞࠸ࠋiNOS mRNA Ⓨ⌧㔞୧⩌㛫࡛ᕪࡣ࡞ࡗࡓࡀ㸪eNOS mRNA Ⓨ⌧㔞ࡣ W1DF ࡛㧗࠸ഴྥࢆ♧ࡋࡓࠋࡇࡢࡇࡽ㸪༸⬊ࡢ⾑⟶ᣑᙇస⏝ W1DF W2DF ࡢ㛫࡛ᕪ ࡀ⏕ࡌ࡚࠸ࡿྍ⬟ᛶࡶ⪃࠼ࡽࢀࡿࠋࡋࡋ࡞ࡀࡽ㸪༸⬊ቨࡢ⾑⟶ࡣෆ༸⬊⭷Ꮡᅾࡋ࡚ ࠾ࡾ㸪ෆ༸⬊⭷⣽⬊࡛Ⓨ⌧ࡋ࡚࠸ࡿ⾑⟶సືᛶᅉᏊࡸ⾑⟶⥙ศᕸࡢ⛬ᗘࢆホ౯ࡋ࡚࠸ࡃ ᚲせࡀ࠶ࡿࠋP4 ྜᡂ㛵ࢃࡿࢫࢸࣟࢻྜᡂ㓝⣲࡛࠶ࡿ P450-scc 3β-HSD mRNA Ⓨ⌧ 㔞࠾ࡼࡧ VEGF164 mRNA Ⓨ⌧㔞ࡣ W1DF ࡛㧗ࡗࡓࠋࡇࢀࡣ㸪༸ᚋᙧᡂࡉࢀࡿ㯤 యࡢ⾑⟶ᙧᡂ࠾ࡼࡧ㯤య⣽⬊ࡢ P4 ྜᡂ⬟ࡀ㧗ࡃ࡞ࡿྍ⬟ᛶࢆ♧ࡋ࡚࠸ࡿࠋࡇࢀࡽࡢࡇ ࡽ㸪W1DF ࡢ LH ࢧ࣮ࢪ᭚㟢క࠺㯤యࡢᛂᛶࡣ㧗࠸ࡇࡀ♧၀ࡉࢀࡓࠋヨ㦂 4 ࡛ࡣ㸪W1DF W2DF ࡢ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᙧែ࠾ࡼࡧᶵ⬟ࢆホ౯ࡋࡓ㸦W1DF 㯤య㸸n㸻6㸪W2DF 㯤య㸸n㸻6㸧 ࠋ㯤య᩿㠃✚㸪㯤య⾑ὶ࠾ࡼࡧ⾑୰ P4 ⃰ᗘࡣ W1DF ༸ᚋᙧᡂࡉࢀࡿ㯤య࡛㧗ࡗࡓࠋࡇࢀࡲ࡛㸪༸๓༸⬊ࡀࡁ࠸ࡑࡢᚋᙧᡂࡉࢀ ࡿ㯤యࡀࡁࡃ࡞ࡿࡇ㸪㯤య୰ᮇ࠾࠸࡚⾑୰ P4 ⃰ᗘࡣ㯤య┤ᚄࡢᙳ㡪ࢆཷࡅࡿࡇ ࡀሗ࿌ࡉࢀ࡚࠸ࡿࠋᮏヨ㦂࡛ࡣ㸪༸๓ࡢ W1DF ࡢᖹᆒ┤ᚄࡀ W2DF ẚ࡚ࡁ࠸ࡇ ࡽ㸪W1DF ࡢ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢ᩿㠃✚ࡀࡁࡃ࡞ࡾ㸪ࡑࡢ⤖ᯝ㸪⾑୰ P4 ⃰ ᗘࡀ㧗ࡃ࡞ࡗࡓ⪃࠼ࡽࢀࡿࠋຍ࠼࡚㸪W1DF ࡢ༸ᚋᙧᡂࡉࢀࡿ㯤యࡣ㸪㯤యᙧᡂ ึᮇ࠾ࡅࡿ㯤యࡢ⾑ὶ㔞ࡀࡁ࠸ࡇ࡛㸪㯤యᙧᡂࡀಁࡉࢀ㯤య᩿㠃✚ࡀࡁࡃ࡞ࡾ㸪 ࡉࡽ⾑ᾮ౪⤥㔞ࡀࡁ࠸ࡇ࡛㯤య⣽⬊ࡢ P4 ྜᡂ⬟࠾ࡼࡧ㯤యࡽ⾑୰ࡢ P4 ᨺฟ ࡢஹ㐍ࡀ⏕ࡌ㸪⾑୰ P4 ⃰ᗘࡀ㧗ࡃ᥎⛣ࡋࡓྍ⬟ᛶࡶ⪃࠼ࡽࢀࡿࠋࡇࢀࡽࡢࡇࡽ㸪 W1DF ࡣ W2DF ẚ࡚㸪༸⬊Ⓨ⫱࠾ࡼࡧ༸๓ᚋࡢᶵ⬟ࡀ㧗࠸ࡇࡀ᥎ ࡉࢀ㸪ࡑࢀ ࡣ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᶵ⬟⮳ࡿࡲ࡛ᙳ㡪ࢆ࠼࡚࠸ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋ 96 ➨ 3 ❶࡛ࡣ㸪⤒⏘㠀Ἢங∵ࢆ⏝࠸࡚ W1DF ࡢⓎ⫱ࡸᶵ⬟ᙳ㡪ࢆ࠼ࡿせᅉࡘ࠸࡚ ㄪࡓࠋヨ㦂 1 ࡛ࡣ㸪ᘏ 44 㢌ࡢ࢘ࢩࢆ⏝࠸ࡓࠋࡲࡎ㸪Day 6 ࡢᖍ༸⬊ࡢᖹᆒ┤ᚄ ⾑୰ P4 ⃰ᗘࡢ┦㛵㛵ಀࢆㄪࡓࠋḟ㸪ᖍ༸⬊ࡢⓎ⫱ࡀ༸ᕢෆࡢ㯤యࡼࡿᙳ㡪ࢆཷ ࡅࡿྍ⬟ᛶࢆ⪃࠼㸪ᖍ༸⬊㯤యࡀྠ୍༸ᕢෆ࡛Ⓨ⫱ࡍࡿඹᏑ⩌ᑐഃࡢ༸ᕢ࡛Ⓨ ⫱ࡍࡿ㠀ඹᏑ⩌ࡢ 2 ࡘศࡅ㸪ࡑࢀࡒࢀࡢ⩌࡛ࡢᖍ༸⬊ࡢᖹᆒ┤ᚄ⾑୰ P4 ⃰ᗘࢆẚ ㍑ࡋࡓࠋࡑࡢ⤖ᯝ㸪Day 6 ࡢᖍ༸⬊ࡢᖹᆒ┤ᚄࡣ⾑୰ P4 ⃰ᗘࡣ┦㛵㛵ಀࡀ࡞࠸ࡇ ࡀ♧ࡉࢀ㸪ࡇࢀࡣඹᏑ⩌㠀ඹᏑ⩌⩌ศࡅࡋࡓሙྜ࡛ࡶྠᵝࡢ⤖ᯝࡀᚓࡽࢀࡓࠋࡇࢀ ࡽࡢࡇࡽ㸪Day 6 ࡢ⾑୰ P4 ⃰ᗘࡣ W1DF ࡢⓎ⫱ᙳ㡪ࢆ࠼ࡿせᅉ࡛ࡣ࡞࠸ࡇࡀ ᫂ࡽ࡞ࡗࡓࠋ୍᪉࡛㸪ᖍ༸⬊ࡢᖹᆒ┤ᚄࡣඹᏑ⩌࡛㠀ඹᏑ⩌ẚ᭷ពࡁ࠸ ࡇࡀ♧ࡉࢀࡓࠋຍ࠼࡚㸪⾑୰ P4 ⃰ᗘࡣ㠀ඹᏑ⩌࡛㧗࠸್ࢆ♧ࡋࡓࠋヨ㦂 2 ࡛ࡣ㸪5 㢌 ࡢ⤒⏘㠀Ἢங∵ࢆ⏝࠸࡚㸪ඹᏑ⩌㸦n㸻5㸧㠀ඹᏑ⩌㸦n㸻5㸧㛫࡛ࡢᖍ༸⬊ࡢᛶ㉁ࢆ ẚ㍑ࡍࡿࡓࡵ㸪Day 0 ࡽ 4 ࡲ࡛ࡢᖍ༸⬊㯤యࡢⓎ⫱ືែ㸪⾑୰ P4 ⃰ᗘ࠾ࡼࡧ Day 4 ࡢ⾑୰ E2 ࠾ࡼࡧ༸⬊ᾮ୰ᛶࢫࢸࣟࢻ࣍ࣝࣔࣥ㸦E2㸪A4㸪P4㸧⃰ᗘࢆ ᐃࡋࡓࠋᖍ༸ ⬊ࡢᖹᆒ┤ᚄࡣඹᏑ⩌࡛㧗ࡃ᥎⛣ࡋࡓࡀ᭷ពᕪࡣ࡞ࡗࡓࠋ㯤యࡢᖹᆒ┤ᚄ࠾ࡼࡧ⾑୰ P4 ⃰ᗘ᥎⛣ᕪࡣ࡞ࡗࡓࠋᖍ༸⬊ࡢᖹᆒ┤ᚄ⾑୰ P4 ⃰ᗘ᥎⛣ᕪࡣ࡞ࡃヨ㦂 1 ␗࡞ࡿ⤖ᯝ࡛࠶ࡗࡓࠋ ࡇࢀࡣヨ㦂 1 ࡛ࡣ Day 6 ᖍ༸⬊ࡢᖹᆒ┤ᚄ⾑୰ P4 ⃰ᗘࢆ ᐃࡋࡓࡓࡵ㸪Ⓨ⫱ᮇ㛫ࡢ㐪࠸ࡼࡿᙳ㡪ࡀ࠶ࡿ⪃࠼ࡽࢀࡿࠋ⾑୰ E2 ⃰ᗘࡣඹᏑ⩌࡛ 㧗࠸ഴྥࢆ♧ࡋࡓࡀ㸪༸⬊ᾮ୰ E2 ࠾ࡼࡧ A4 ⃰ᗘᕪࡣ࡞ࡗࡓࠋࡋࡋ࡞ࡀࡽ㸪༸⬊ ᾮ୰ P4 ⃰ᗘࡣඹᏑ⩌࡛㧗ࡗࡓࠋ㯤యྠ୍༸ᕢෆ࡛ඹᏑࡍࡿ W1DF ࡣ㸪༸ᕢෆ༢⊂ ࡛Ꮡᅾࡍࡿ W1DF ẚ࡚㸪ᖹᆒ┤ᚄ㸪༸ᕢࡢ⾑ὶ㔞࠾ࡼࡧ༸⬊ቨ⾑ὶ㠃✚ࡀࡁ࠸ ࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿࡇࡽ㸪ᮏヨ㦂࠾࠸࡚ࡶඹᏑ⩌࡛ࡢ༸ᕢ࠾ࡼࡧ༸⬊ࡢ⾑ὶ 㔞ࡀࡁ࠸ࡇࡀ᥎ ࡉࢀ㸪㓟⣲㸪ᰤ㣴࠾ࡼࡧ࣍ࣝࣔࣥࡀࡼࡾ༸⬊౪⤥ࡉࢀࡿࡇ࡛ ᖹᆒ┤ᚄࡀࡁࡃ࡞ࡾ㸪༸⬊࡛⏘⏕ࡉࢀࡓ E2 ࡀࡼࡾ㌟ᚠ⎔୰ᨺฟࡉࢀࡿࡇ࡛⾑୰ E2 ⃰ᗘࡀ㧗ࡃ࡞ࡗࡓ⪃࠼ࡽࢀࡿࠋ୍᪉㸪⾑୰ P4 ⃰ᗘࡀඹᏑ⩌࡛పࡗࡓࡀ㸪⾑୰ P4 97 ⃰ᗘᕪࡀ⏕ࡌࡓཎᅉࡣᮏ◊✲࡛ࡣ᫂ࡽ࡛ࡁ࡞ࡗࡓࠋ㯤యᶵ⬟ࡢホ౯ࡸඹᏑࡍࡿ ༸⬊ࡀ㯤యࡢᶵ⬟ᙳ㡪ࢆ࠼࡚࠸ࡿྍ⬟ᛶࡶྵࡵ࡚㸪᭦࡞ࡿ◊✲ࢆ⾜࠺ᚲせࡀ࠶ࡿ ⪃࠼ࡽࢀࡿࠋ༸⬊ᾮ୰ E2 ࠾ࡼࡧ A4 ⃰ᗘᕪ␗ࡀ࡞ࡗࡓࡇࡽ㸪ඹᏑ⩌㠀ඹᏑࡢ 㛫࡛༸⬊ᶵ⬟ᕪ␗ࡣ࡞࠸⪃࠼ࡽࢀࡿࡓࡵ㸪༸⬊ᾮ P4 ⃰ᗘࡢᕪ␗ࡣ㏆㞄ࡢ㯤య࡛⏘⏕ ࡉࢀࡓ㧗⃰ᗘࡢ P4 ࡀ༸ᕢෆࡢ⾑⟶ࢆࡋ࡚ᖍ༸⬊฿㐩ࡋධࡋࡓ⪃࠼ࡽࢀࡿࠋࡋ ࡋ࡞ࡀࡽ㸪ࡇࡢⅬ㛵ࡋ࡚ࡶ᥎ ࡢᇦࢆ㉺࠼࡞࠸ࡓࡵ㸪༸⬊ᶵ⬟ࡢᕪ␗ࡀᏑᅾࡍࡿྍ ⬟ᛶࡶྵࡵ᭦࡞ࡿ◊✲ࢆ⾜࠺ᚲせࡀ࠶ࡿ⪃࠼ࡽࢀࡿࠋ➨ 3 ❶࡛ࡣ㸪W1DF ࡢᛶ㉁ᙳ 㡪ࢆཬࡰࡍせᅉࡋ࡚㸪ྠ୍༸ᕢෆ࡛㯤యඹᏑࡍࡿྰ࠸࠺⨨㛵ಀࡢせᅉࡀ ࡁ࠸ࡇࡀ᫂ࡽ࡞ࡗࡓࠋ ➨ 4 ❶࡛ࡣ㸪➨ 3 ❶࡛᫂ࡽ࡞ࡗࡓ⌧㇟ࡀ㸪⤒⏘Ἢங∵࠾࠸࡚ࡶほᐹࡉࢀࡿ⌧㇟ ࡛࠶ࡿㄪࡓࠋࡲࡓ㸪㐣ཤࡢሗ࿌ࡽ㸪㯤యࡀᏑᅾࡍࡿ༸ᕢྠഃࡢᏊᐑゅࡢᏊᐑෆ ⭷࡛ࡢ⤌⧊୰ P4 ⃰ᗘࡣᑐഃࡢᏊᐑෆ⭷ẚ㍑ࡋ࡚㧗࠸ࡇ㸪༸๓༸⬊ࡀᏑᅾࡍࡿഃ ࡢ༸⟶ᾮ୰ࡢ E2 ⃰ᗘࡀ㧗࠸ࡇࡽ㸪༸ᕢྠഃࡢᏊᐑゅࡣᒁᡤⓗ࡞ࡘ࡞ࡀࡾࡀ࠶ࡿ ࡇࡀ⪃࠼ࡽࢀࡿࠋ࢘ࢩࡣ༸ࡋࡓഃࡢᏊᐑゅ࡛㧗⋡╔ᗋࡋዷፎ⮳ࡿࡇࡀ▱ࡽࢀ ࡚࠾ࡾ㸪ேᕤᤵ⢭ᚋࡢ W1DF ࡀ㯤యඹᏑࡋࡓሙྜ㸪ఱࡽࡢᒁᡤⓗ࡞స⏝ࡼࡾ㸪ࡑ ࡢᚋࡢཷ⫾ᛶᙳ㡪ࢆ࠼ࡿྍ⬟ᛶࡀ࠶ࡿࠋࡑࡢࡓࡵ㸪ேᕤᤵ⢭ᚋࡢ W1DF 㯤యࡢ ⨨㛵ಀࢆㄪ㸪ࡑࡢᚋࡢཷ⫾⋡ࢆẚ㍑ࡋࡓࠋヨ㦂 1 ࡛ࡣ㸪༸☜ㄆ᪥ࢆ Day 0 ࡋ㸪Day 2㸪4㸪6㸪8 ࠾ࡼࡧ 10 ᖍ༸⬊࠾ࡼࡧ㯤యࡢᖹᆒ┤ᚄࢆィ ࡋ㸪ࡉࡽ᥇⾑ࢆ⾜࠸⾑ ୰ P4 ⃰ᗘࡢ ᐃࢆ⾜࠸㸪ඹᏑ⩌㠀ඹᏑ⩌㛫࡛ẚ㍑ࡋࡓࠋᖍ༸⬊ࡢᖹᆒ┤ᚄࡣඹᏑ⩌ ࡛ Day 6 ࡽࡁࡃ࡞ࡾ㸪Day 8 ࡣ㠀ඹᏑẚ㍑ࡋ࡚᭷ពࡁࡃ࡞ࡗࡓࠋ㯤యࡢᖹᆒ ┤ᚄ࠾ࡼࡧ⾑୰ P4 ⃰ᗘࡢ᥎⛣ࡣ୧⩌㛫࡛ᕪࡣ࡞ࡗࡓࠋ⾑୰ P4 ⃰ᗘᕪࡀ࡞࠸ࡶ㛵 ࢃࡽࡎ㸪ඹᏑ⩌࡛ࡢᖍ༸⬊ࡢᖹᆒ┤ᚄࡀࡁࡃ࡞ࡗࡓࡇࡽ㸪➨ 3 ❶࡛⪃ᐹࡋࡓࡼ ࠺㸪㯤యࡀඹᏑࡍࡿ༸ᕢ࡛Ⓨ⫱ࡍࡿᖍ༸⬊ࡣ㸪༸ᕢࡢ⾑ὶࡀࡁࡃ࡞ࡿࡇ࡛㓟 ⣲㸪ᰤ㣴࠾ࡼࡧ࣍ࣝࣔࣥ࡞ࡀࡼࡾ౪⤥ࡉࢀࡿࡇ࡛Ⓨ⫱ࡀಁࡉࢀࡓ⪃࠼ࡽࢀࡿࠋヨ 98 㦂 2 ࠾࠸࡚㸪ேᕤᤵ⢭ᚋࡢཷ⫾⋡ࡣ⤒⏘Ἢங∵࠾ࡼࡧᮍ⤒⏘∵ࡶඹᏑ⩌࡛పୗࡍ ࡿࡇࡀ᫂ࡽ࡞ࡗࡓࠋࡉࡽ㸪ࡇࡢ⌧㇟ࡣ㸪Ꮨ⠇㸪ศፔᚋ᪥ᩘ㸪⏘ḟࡢᙳ㡪ࢆཷࡅ ࡞࠸ࡇࡶ᫂ࡽ࡞ࡗࡓࠋࡇࡢ⌧㇟ࡣ㸪W1DF ࡀ⦾Ṫ⏕⌮ᙳ㡪ࢆ࠼ࡿࡇࢆ᫂ࡽ ࡋࡓึࡵ࡚ࡢሗ࿌࡛࠶ࡿࠋ➨ 3 ❶ࡢヨ㦂 2 ࠾࠸࡚㸪W1DF ࡣ༸⬊ෆ࡛༑ศ㔞ࡢ E2 ࢆ⏘⏕ࡋ⾑୰ E2 ⃰ᗘᙳ㡪ࢆ࠼ࡿࡇࢆ♧ࡋࡓࡀ㸪࢘ࢩ࠾࠸࡚༸ᕢࡽࡢ⾑⟶ࡀ༸ ⟶Ꮚᐑゅඛ➃㐃⤖ࡋ࡚࠸ࡿࡇࡀศࡗ࡚࠾ࡾ㸪ྠഃࡢ༸⟶ࡸᏊᐑゅᑐࡋ࡚ࡼࡾ 㧗⃰ᗘ࡛ᙳ㡪ࢆ࠼ࡿࡇࡀ⪃࠼ࡽࢀࡿࠋࡲࡓ E2 ࡣᏊᐑෆ⭷⣽⬊స⏝ࡋ࡚ E2 ࠾ࡼࡧ P4 ཷᐜయⓎ⌧ᙳ㡪ࢆ࠼ࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿࡓࡵ㸪ཷ⢭༸ࡀᏑᅾࡍࡿ༸⟶ࡸᏊᐑ ゅෆ⭷ᑐࡋ࡚㧗⃰ᗘࡢ E2 ࡀస⏝ࡍࡿࡇ࡛㸪⬇ࡢⓎ⫱ࡸཷᐜ⬟ᑐࡋ࡚ᙳ㡪ࢆ࠼ ࡿྍ⬟ᛶࡀ࠶ࡿࠋࡋࡋ࡞ࡀࡽ㸪ᮏヨ㦂࡛ࡣཷ⫾ᛶᕪ␗ࢆࡶࡓࡽࡍᶵᵓࡲ࡛ࢆ♧ࡍࡇ ࡣ࡛ࡁ࡞ࡗࡓࠋᚋࡣཷ⫾ᛶపୗࡢ࣓࢝ࢽࢬ࣒ཷ⫾ᛶྥୖࡲ࡛ࢆど㔝ධࢀࡓ◊ ✲ࢆ⾜ࡗ࡚࠸ࡃᚲせࡀ࠶ࡿ⪃࠼ࡽࢀࡿࠋ ௨ୖࡼࡾ㸪➨ 2 ❶ࡽ➨ 4 ❶ࡲ࡛⾜ࡗࡓ◊✲ࡽ㸪W1DF ࡢⓎ⫱ືែ㸪ᙧែ㸪༸⬊ ቨ⾑ὶ㸪༸๓ᚋࡢ㢛⢏ᒙ⣽⬊ࡢᛶ㉁࠾ࡼࡧ༸ᚋᙧᡂࡉࢀࡿ㯤యࡢᛶ㉁ࢆ W2DF ẚ㍑ࡋࡓ⤖ᯝ㸪άຊࡢ㧗࠸༸⬊࡛࠶ࡿࡇࡀ᫂ࡽ࡞ࡾ㸪W1DF ࡀⓎ⫱ࡍࡿᮇ㛫ࡢప ࠸⾑୰ P4 ⃰ᗘࡀࡑࡢཎᅉࢆࡶࡓࡽࡋࡓ᥎ᐹࡉࢀࡿࠋࡉࡽ㸪W1DF ࡘ࠸࡚㸪㯤య ྠ୍༸ᕢ࡛ඹᏑࡍࡿሙྜ༢⊂࡛ᏑᅾࡋඹᏑࡋ࡞࠸ሙྜ࡛ࡣ㸪ᖍ༸⬊ࡢᛶ㉁ࡀ␗࡞ࡿ ࡇࡀ♧ࡉࢀ㸪༸ᕢࡢ⾑ὶࡢ㐪࠸ࡀࡑࡢᕪ␗ࢆࡶࡓࡽࡍ᥎ ࡉࢀࡓࠋࡲࡓ㸪㯤య ༸⬊ࡀඹᏑࡍࡿࡇࡀேᕤᤵ⢭ᚋࡢཷ⫾⋡ࢆపୗࡉࡏࡓࡇࡽ㸪࢘ࢩࡢ⦾Ṫ⏕⌮࠾ ࡅࡿ W1DF ࡢ᪂ࡓ࡞ᙺ㛵ࡍࡿ▱ぢࡀᚓࡽࢀࡓࠋ 99 ㅰ㎡ ᮏ✏ࢆ⤊࠼ࡿ⮫ࡳ㸪⤊ጞᚚ᠓⠜࡞ࡿᚚᣦᑟࢆ㈷ࡾࡲࡋࡓᖏᗈ␆⏘Ꮫ␆⏘Ꮫ⛉ ⮫ᗋ⋇་Ꮫ◊✲㒊㛛 ᯇᇶ⣧ᩍᤵ㸪⩚⏣┿ᝅຓᩍᚰࡽᣏㅰࡋ㸪ᮏㄽᩥసᡂ ࠶ࡓࡾᚚᰯ㜀㡬ࡁࡲࡋࡓᖏᗈ␆⏘Ꮫ ᐑཎ㑻ᩍᤵ㸪ᒾᡭᏛ ᒃᅾᐙ⩏ᩍ ᤵ㸪ᮾி㎰ᕤᏛ ⏣୰▱ᕫᩍᤵ㸪ᒱ㜧Ꮫ ᮧ℩ဴ☻ᩍᤵ῝ㅰ࠸ࡓࡋࡲࡍࠋ ࡲࡓ㸪◊✲㐙⾜࠶ࡓࡾ㸪ᚚ༠ຊ㸪ᚚຓゝ㡬ࡁࡲࡋࡓ㸪ᖏᗈ␆⏘Ꮫ ᐑᮏ᫂ኵᩍ ᤵ㸪ΎỈ㝯ᩍᤵ㸪ⱴ㔝ග⠊ㅮᖌ῝ㅰ࠸ࡓࡋࡲࡍࠋ ࡑࡋ࡚㸪ᐇ㦂༠ຊࡋ࡚ࡃࡔࡉࡗࡓ⋇་⮫ᗋ⦾ṪᏛ◊✲ᐊࡢᏛ㝔⏕㸪Ꮫ㒊Ꮫ⏕ ࡢⓙᵝឤㅰ࠸ࡓࡋࡲࡍࠋ ᭱ᚋ㸪Ꮫ⏕⏕άࢆ㛗ࡁࢃࡓࡾᨭ࠼࡚࠸ࡓࡔ࠸ࡓ୧ぶ᭱ࡢឤㅰࡢពࢆ⏦ࡋ ㏙ࡲࡍࠋ 100 ཧ⪃ᩥ⊩ 1. 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