1997 年度実績報告書

アラキドン酸カスケードと病的動態

研究課題

研究課題/領域番号	07557171
研究機関	東北大学
研究代表者	水柿道直東北大学, 医学部・附属病院, 教授 (60004595)
研究分担者	山本尚三徳島大学, 医学部, 教授 (50025607) 室田誠逸東京医科歯科大学, 歯学部, 教授 (50072989) 清水孝雄東京大学, 医学部, 教授 (80127092) 尾股健東北大学, 医学部・附属病院, 講師 (50194634) 大内和雄東北大学, 薬学部, 教授 (20006357)
キーワード	プロスタグランジン / ロイコトリエン / シクロオキシゲナーゼ / GC / MS / トロンボキサン / プロスタサイクリン / 炎症 / 病態
研究概要	年度の研究により、アラキドン酸カスケードに関わる諸因子との関連性について以下の点を明らかにした。 (1)プロスタグランジンの産生と病態;高脂血症患者に於ける血清脂質及び尿中prostacyclin(PGI_2)、thromboxaneA_2(TXA_2)代謝産物、8-isoprostane排泄量を測定し、高脂血症患者におけるTXA_2/PGI_2産生不均衡とその血管合併症への関与が示唆された。またイコサペント酸エチル投与により、PGI_2、TXA_2及び8-isoprostaneがその効果に関与することが示唆された。さらに、異常妊娠例(妊娠性糖尿病例、子宮内胎児発育遅延例など)や糖尿病患者では、TXA_2/PGI_2産生不均衡が生じていることが示された。 (2)シクロオキシゲナーゼの変動;急性炎症モデルに於けるCOX-2阻害薬の抗炎症作用はCOX-2を介するPGE_2産生阻害であること、更にデキサメタゾンの血漿滲出抑制作用はPLA_2及びCOX-2の発現抑制は重要でないことが示された。血管新生モデルを用いての検討では、COX-1でなくCOX-2がPG生成を介して血管新生を増強していることが示され、誘導されたVEGFが実際にこのモデルの血管新生に役割を持つことが判明した。また、COX酵素がCOX活性とperoxidase活性を合わせ持っていることを利用して、生細胞系でCOX活性を個々の細胞で測定できる系を確立し、内因性アラキドン酸を利用できるのはCOX-2のみであること、NOはCOX-2活性を有意に抑制することを明らかにした。COX-1はヒト巨核芽球性白血病細胞CMKでホルボールエステルで誘導され、血小板では認められないPGD合成酵素の造血器型がCMK細胞に存在することが示された。COX-2はマウス骨芽細胞MC3T3-E1でTNF_αによって著明に誘導され、その転写調節因子としてNFkBとNFIL-6の関与が明らかとなった。 (3)ロイコトリエン(LT)の定量と機能;LTB_4の定量分析には、LTB_4のモノクローン抗体を用いた放射免疫測定法、尿サンプルを用いる簡便な酵素免疫測定法、ESIイオン源を装着したLC/MS微量定量分析をそれぞれ確立した。また、喘息患者では謙常人に比べて尿中LTE_4、11β-PGF_<2α>とも有意に高値を示し、アスピリン喘息患者では尿中LTE_4が非常に高値を示し、肺気腫患者では11β-PGF_<2α>が有意に高値を示した。新規抗糖尿病薬troglitazoneには抗原刺激によるLT産生を抑制する作用があることをラット肥満細胞株であるRBL-2H3細胞を用いて明らかとなった。またLTB_4受容体のクローニングに成功し、その性状を明らかにした。CHO細胞に発現させた場合、LTB_4はアデニル酸シクラーゼ活性の阻害、細胞内カルシウム濃度の上昇を引き起こし、また、LTB_4の遊走活性への関与が明らかとなった。

研究成果

(37件)

すべてその他

すべて文献書誌 (37件)

[文献書誌] Takano,S.: "Determination of ascorbic acid in human vitreous humor by high-performance liquid chromatography with UV detection." Current.Eye Research. 16(6). 589-594 (1997)
[文献書誌] Hishinuma,T.: "Microdetermination of the thro_mboxane B_3 by gaschromatography-selected ion moni ng using [^<18>O]thro_mboxane B_3 as an internal standard" Prostaglandins. 53. 381-394 (1997)
[文献書誌] Nishikawa,M.: "Effect of eicosapentaenoic acid (EPA) on prostacyclin production in diabetics:GC-MS analy_sis of PG_<12> and PGl_3 levels" Meth.Find.Exp.Clin.Pharmacol. 19(6). 429-433 (1997)
[文献書誌] Suzuki,H.: "Expressions of P-selectin-and HSP72-like immunoreactivites in rat brain after transient middle cerebral artery occlusion" Brain Research. 759. 321-329 (1997)
[文献書誌] Suzuki,H.: "Postischemic expression of P-selectin immunoreactivity in rat brain" Neuroscience Letters. 228. 151-154 (1997)
[文献書誌] Deguchi,M.: "Detection of human papillomavirus type 18 DNA in squamous cell carcinoma developing from bowen‘s disease of the face" Eur J Dermatol. 7. 487-488 (1997)
[文献書誌] Kume,K.: "Microplate chromatography assay foracetyl-CoA:lysoplatelet-activating factor acetyltransferase." Anal.Biochem.246. 118-122 (1997)
[文献書誌] Ishii,I.: "Alanine exchanges of polar amino acids in the transmembrane domains of a platelet-activating factor receptor generate both constitutively active and inactive mutants." J.Biol.Chem.272. 7846-7854 (1997)
[文献書誌] Ogita,T.: "Lysophosphatidylcholine transduces Ca2_+ signaling via the platelet-activating factor receptor in macrophages." Am.J.Physiol.272. H17-H24 (1997)
[文献書誌] Ito,K.: "Molecular cloning of canalicular multispecific organic anion transporter defective in EHBR." Am.J.Physiol. 272. G16-G22 (1997)
[文献書誌] Mori,M.: "Differential expression of somatostatin receptors in the rat eye SSTR4 is intensely expressed in the iris/ciliary body." Neurosci.Lett.223. 185-188 (1997)
[文献書誌] Yokomizo,T.: "A G-protein coupled receptor for leukotriene B_4 that mediates chemotaxis." Nature. 387. 620-624 (1997)
[文献書誌] Mizugaki,M.: "Alteration of DNA methylation levels in MRL lupus mice" Clinical and Experimental Immunology. 110. 265-269 (1997)
[文献書誌] Mizugaki,M.: "The influence of general anesthesia on brain distribution and kinetics of [^<11>C]methamphetamine in monkey" CYRIC Ann.Rep.(in press).
[文献書誌] Deguchi,M.: "Detection of human papillomavirus type 33 DNA in extragenital bowen‘s disease with the polmerase chain reaction" Arch.Dermatol.Res. (in press).
[文献書誌] Sato,H.: "Pharmaceutical properties of nifedipine from a ground mixture with nifedipin,casein,magnesium silicate and cellulose acetate phthalate" Jpn.J.Hosp.Pharm. 23(2). 101-107 (1997)
[文献書誌] Yamanobe,S.: "Analysis of urinary 11-dehydrothromboxane B_2 in patients with occluded retinal vein using GC/SIM" Prostaglandins leukotriens and essential fatty acids. 58(1). 65-68 (1998)
[文献書誌] Ishii,S.: "Bronchial hyperreactivity,increased endotoxin lethality,and melanocytic tumorigenesis in transgenic mice overexpressing platelet-activating factor receptor." EMBO J.16. 133-142 (1997)
[文献書誌] N,Uozumi.: "Role of cytosolic phospholipase A2 in allergic response and parturition." Nature. 390. 618-622 (1997)
[文献書誌] Liu,B.: "Prolonged activation of phospholipase D in Chinese hamster ovary cells expressing platelet-activating factor receptor lackingcytoplasmic C-terminal." Biochem.J.327. 239-244 (1997)
[文献書誌] Hoshino,M.: "Leukotriene D4 activates mitogen-activated protein kinase through a protein kinase C alpha-Raf-1-dependent pathway in human THP-1 cells." J.Biol.Chem.273. 4878-4882 (1998)
[文献書誌] Ishii,I.: "Agonis-induced sequestration,recycling and resensitization of platelet-activating factor receptor-role of cytoplasmic tail phosphorylation in each process." J.Biol.Chem.(in press).
[文献書誌] 山本尚三: "シクロオキシゲナーゼ-1と-2の基礎" 現代医療. 29. 2297-2302 (1997)
[文献書誌] Ueda,N.: "Induction of cyclooxygenase-1 in a human megakaryoblastic cell line (CMK) differentiated by phorbol ester." Biochim.Biophys.Acta. 1344. 103-110 (1997)
[文献書誌] Kume,K.: "Growth stimulation,inhibition and suppressin of oncogenic transformation of NRK cells by platelet-activating factor." J.Biol.Chem. 272. 22898-22904 (1997)
[文献書誌] Liu,B.: "Prolonged activation of phospholipase D in Chinese hamster ovary cells expressing PAF receptor lacking cytoplasmic carboxy-terminal tail." Biochem.J.(in press).
[文献書誌] Honda,Z.: "Roles of C-terminal Src kinase (CsK) in the initiation and the termination of FceRl-mediated signaling." J.Biol.Chem. 272. 25753-25760 (1997)
[文献書誌] Endoh,H.: "Rapid activation of MAP kinase by estrogen in the bone cell line." Biochem.Biophys.Res.Commun. 235. 99-102 (1997)
[文献書誌] Kume,K.: "cDNA cloning and expression of murine1-acyl-sn-glyserol-3-phosphate acyltransferase." Biochem.Biophys.Res.Commun.237. 663-666 (1997)
[文献書誌] Tabuchi,S.: "Lipid mediators modulate NMDA receptor currents in a Xenopus oocyte expression system." Neurosci.Lett.237. 13-16 (1997)
[文献書誌] Mahmud,I.: "Prostaglandin D synthase in human megakaryoblastic cells." J.Biol.Chem.272. 28263-28266 (1997)
[文献書誌] Yan SP: "Eicosapentaenoic acid pretreatment attenuated the cell proliferation response to vascular endothelial growth factor via down-regulation of KDR/Flk-1 expression in bovine vascular endothelial cells." Journal of Cellular Physiology. (in press).
[文献書誌] Shitashige M.: "Different substrate utilization between prostaglandin endoperoxide H synthase-1 and -2 in NIH3T3 fibroblasts." Biochimica et Biophysica Acta. 1389. 57-66 (1998)
[文献書誌] Kitamura Y.: "Effect of IL-6 on tumor cell invasion of vascular endothelial monolayers." Surgery Today. 27(6). 534-541 (1997)
[文献書誌] Yamashita H.: "Growth/dufferentiation factor-5 induces angiogenesis in vivo." Experimantal Cell Research.235(1). 218-226 (1997)
[文献書誌] Iwamoto M.: "A biotinylated perfringolysin O derivative : a new probe for detection of cell surface cholesterol." Biochimica et Biophysica Acta.1327(2). 222-23 (1997)
[文献書誌] Morita,I.: "Enhancement of membrane fluidity in cholesterol-poor endothelial cells pre-treated with simvastin." Endothelium. 5(2). 107-113 (1997)

1997 年度 実績報告書

アラキドン酸カスケードと病的動態

研究代表者

水柿 道直 東北大学, 医学部・附属病院, 教授 (60004595)

研究成果

[文献書誌] Takano,S.: "Determination of ascorbic acid in human vitreous humor by high-performance liquid chromatography with UV detection." Current.Eye Research. 16(6). 589-594 (1997)

[文献書誌] Hishinuma,T.: "Microdetermination of the thro_mboxane B_3 by gaschromatography-selected ion moni ng using [^<18>O]thro_mboxane B_3 as an internal standard" Prostaglandins. 53. 381-394 (1997)

[文献書誌] Nishikawa,M.: "Effect of eicosapentaenoic acid (EPA) on prostacyclin production in diabetics:GC-MS analy_sis of PG_<12> and PGl_3 levels" Meth.Find.Exp.Clin.Pharmacol. 19(6). 429-433 (1997)

[文献書誌] Suzuki,H.: "Expressions of P-selectin-and HSP72-like immunoreactivites in rat brain after transient middle cerebral artery occlusion" Brain Research. 759. 321-329 (1997)

[文献書誌] Suzuki,H.: "Postischemic expression of P-selectin immunoreactivity in rat brain" Neuroscience Letters. 228. 151-154 (1997)

[文献書誌] Deguchi,M.: "Detection of human papillomavirus type 18 DNA in squamous cell carcinoma developing from bowen‘s disease of the face" Eur J Dermatol. 7. 487-488 (1997)

[文献書誌] Kume,K.: "Microplate chromatography assay foracetyl-CoA:lysoplatelet-activating factor acetyltransferase." Anal.Biochem.246. 118-122 (1997)

[文献書誌] Ishii,I.: "Alanine exchanges of polar amino acids in the transmembrane domains of a platelet-activating factor receptor generate both constitutively active and inactive mutants." J.Biol.Chem.272. 7846-7854 (1997)

[文献書誌] Ogita,T.: "Lysophosphatidylcholine transduces Ca2_+ signaling via the platelet-activating factor receptor in macrophages." Am.J.Physiol.272. H17-H24 (1997)

[文献書誌] Ito,K.: "Molecular cloning of canalicular multispecific organic anion transporter defective in EHBR." Am.J.Physiol. 272. G16-G22 (1997)

[文献書誌] Mori,M.: "Differential expression of somatostatin receptors in the rat eye SSTR4 is intensely expressed in the iris/ciliary body." Neurosci.Lett.223. 185-188 (1997)

[文献書誌] Yokomizo,T.: "A G-protein coupled receptor for leukotriene B_4 that mediates chemotaxis." Nature. 387. 620-624 (1997)

[文献書誌] Mizugaki,M.: "Alteration of DNA methylation levels in MRL lupus mice" Clinical and Experimental Immunology. 110. 265-269 (1997)

[文献書誌] Mizugaki,M.: "The influence of general anesthesia on brain distribution and kinetics of [^<11>C]methamphetamine in monkey" CYRIC Ann.Rep.(in press).

[文献書誌] Deguchi,M.: "Detection of human papillomavirus type 33 DNA in extragenital bowen‘s disease with the polmerase chain reaction" Arch.Dermatol.Res. (in press).

[文献書誌] Sato,H.: "Pharmaceutical properties of nifedipine from a ground mixture with nifedipin,casein,magnesium silicate and cellulose acetate phthalate" Jpn.J.Hosp.Pharm. 23(2). 101-107 (1997)

[文献書誌] Yamanobe,S.: "Analysis of urinary 11-dehydrothromboxane B_2 in patients with occluded retinal vein using GC/SIM" Prostaglandins leukotriens and essential fatty acids. 58(1). 65-68 (1998)

[文献書誌] Ishii,S.: "Bronchial hyperreactivity,increased endotoxin lethality,and melanocytic tumorigenesis in transgenic mice overexpressing platelet-activating factor receptor." EMBO J.16. 133-142 (1997)

[文献書誌] N,Uozumi.: "Role of cytosolic phospholipase A2 in allergic response and parturition." Nature. 390. 618-622 (1997)

[文献書誌] Liu,B.: "Prolonged activation of phospholipase D in Chinese hamster ovary cells expressing platelet-activating factor receptor lackingcytoplasmic C-terminal." Biochem.J.327. 239-244 (1997)

[文献書誌] Hoshino,M.: "Leukotriene D4 activates mitogen-activated protein kinase through a protein kinase C alpha-Raf-1-dependent pathway in human THP-1 cells." J.Biol.Chem.273. 4878-4882 (1998)

[文献書誌] Ishii,I.: "Agonis-induced sequestration,recycling and resensitization of platelet-activating factor receptor-role of cytoplasmic tail phosphorylation in each process." J.Biol.Chem.(in press).

[文献書誌] 山本尚三: "シクロオキシゲナーゼ-1と-2の基礎" 現代医療. 29. 2297-2302 (1997)

[文献書誌] Ueda,N.: "Induction of cyclooxygenase-1 in a human megakaryoblastic cell line (CMK) differentiated by phorbol ester." Biochim.Biophys.Acta. 1344. 103-110 (1997)

[文献書誌] Kume,K.: "Growth stimulation,inhibition and suppressin of oncogenic transformation of NRK cells by platelet-activating factor." J.Biol.Chem. 272. 22898-22904 (1997)

[文献書誌] Liu,B.: "Prolonged activation of phospholipase D in Chinese hamster ovary cells expressing PAF receptor lacking cytoplasmic carboxy-terminal tail." Biochem.J.(in press).

[文献書誌] Honda,Z.: "Roles of C-terminal Src kinase (CsK) in the initiation and the termination of FceRl-mediated signaling." J.Biol.Chem. 272. 25753-25760 (1997)

[文献書誌] Endoh,H.: "Rapid activation of MAP kinase by estrogen in the bone cell line." Biochem.Biophys.Res.Commun. 235. 99-102 (1997)

[文献書誌] Kume,K.: "cDNA cloning and expression of murine1-acyl-sn-glyserol-3-phosphate acyltransferase." Biochem.Biophys.Res.Commun.237. 663-666 (1997)

[文献書誌] Tabuchi,S.: "Lipid mediators modulate NMDA receptor currents in a Xenopus oocyte expression system." Neurosci.Lett.237. 13-16 (1997)

[文献書誌] Mahmud,I.: "Prostaglandin D synthase in human megakaryoblastic cells." J.Biol.Chem.272. 28263-28266 (1997)

[文献書誌] Yan SP: "Eicosapentaenoic acid pretreatment attenuated the cell proliferation response to vascular endothelial growth factor via down-regulation of KDR/Flk-1 expression in bovine vascular endothelial cells." Journal of Cellular Physiology. (in press).

[文献書誌] Shitashige M.: "Different substrate utilization between prostaglandin endoperoxide H synthase-1 and -2 in NIH3T3 fibroblasts." Biochimica et Biophysica Acta. 1389. 57-66 (1998)

[文献書誌] Kitamura Y.: "Effect of IL-6 on tumor cell invasion of vascular endothelial monolayers." Surgery Today. 27(6). 534-541 (1997)

[文献書誌] Yamashita H.: "Growth/dufferentiation factor-5 induces angiogenesis in vivo." Experimantal Cell Research.235(1). 218-226 (1997)

[文献書誌] Iwamoto M.: "A biotinylated perfringolysin O derivative : a new probe for detection of cell surface cholesterol." Biochimica et Biophysica Acta.1327(2). 222-23 (1997)

[文献書誌] Morita,I.: "Enhancement of membrane fluidity in cholesterol-poor endothelial cells pre-treated with simvastin." Endothelium. 5(2). 107-113 (1997)

1997 年度実績報告書

水柿道直東北大学, 医学部・附属病院, 教授 (60004595)