1997 Fiscal Year Annual Research Report

アラキドン酸カスケードと病的動態

Research Project

Project/Area Number	07557171
Research Institution	Tohoku University
Principal Investigator	水柿道直東北大学, 医学部・附属病院, 教授 (60004595)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	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
Research Abstract	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。

Research Products
(37 results)

All Other

All Publications (37 results)

[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Suzuki,H.: "Postischemic expression of P-selectin immunoreactivity in rat brain" Neuroscience Letters. 228. 151-154 (1997)
[Publications] 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)
[Publications] Kume,K.: "Microplate chromatography assay foracetyl-CoA:lysoplatelet-activating factor acetyltransferase." Anal.Biochem.246. 118-122 (1997)
[Publications] 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)
[Publications] Ogita,T.: "Lysophosphatidylcholine transduces Ca2_+ signaling via the platelet-activating factor receptor in macrophages." Am.J.Physiol.272. H17-H24 (1997)
[Publications] Ito,K.: "Molecular cloning of canalicular multispecific organic anion transporter defective in EHBR." Am.J.Physiol. 272. G16-G22 (1997)
[Publications] 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)
[Publications] Yokomizo,T.: "A G-protein coupled receptor for leukotriene B_4 that mediates chemotaxis." Nature. 387. 620-624 (1997)
[Publications] Mizugaki,M.: "Alteration of DNA methylation levels in MRL lupus mice" Clinical and Experimental Immunology. 110. 265-269 (1997)
[Publications] Mizugaki,M.: "The influence of general anesthesia on brain distribution and kinetics of [^<11>C]methamphetamine in monkey" CYRIC Ann.Rep.(in press).
[Publications] Deguchi,M.: "Detection of human papillomavirus type 33 DNA in extragenital bowen‘s disease with the polmerase chain reaction" Arch.Dermatol.Res. (in press).
[Publications] 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)
[Publications] 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)
[Publications] Ishii,S.: "Bronchial hyperreactivity,increased endotoxin lethality,and melanocytic tumorigenesis in transgenic mice overexpressing platelet-activating factor receptor." EMBO J.16. 133-142 (1997)
[Publications] N,Uozumi.: "Role of cytosolic phospholipase A2 in allergic response and parturition." Nature. 390. 618-622 (1997)
[Publications] 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)
[Publications] 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)
[Publications] 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).
[Publications] 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)
[Publications] Ueda,N.: "Induction of cyclooxygenase-1 in a human megakaryoblastic cell line (CMK) differentiated by phorbol ester." Biochim.Biophys.Acta. 1344. 103-110 (1997)
[Publications] Kume,K.: "Growth stimulation,inhibition and suppressin of oncogenic transformation of NRK cells by platelet-activating factor." J.Biol.Chem. 272. 22898-22904 (1997)
[Publications] Liu,B.: "Prolonged activation of phospholipase D in Chinese hamster ovary cells expressing PAF receptor lacking cytoplasmic carboxy-terminal tail." Biochem.J.(in press).
[Publications] 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)
[Publications] Endoh,H.: "Rapid activation of MAP kinase by estrogen in the bone cell line." Biochem.Biophys.Res.Commun. 235. 99-102 (1997)
[Publications] Kume,K.: "cDNA cloning and expression of murine1-acyl-sn-glyserol-3-phosphate acyltransferase." Biochem.Biophys.Res.Commun.237. 663-666 (1997)
[Publications] Tabuchi,S.: "Lipid mediators modulate NMDA receptor currents in a Xenopus oocyte expression system." Neurosci.Lett.237. 13-16 (1997)
[Publications] Mahmud,I.: "Prostaglandin D synthase in human megakaryoblastic cells." J.Biol.Chem.272. 28263-28266 (1997)
[Publications] 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).
[Publications] Shitashige M.: "Different substrate utilization between prostaglandin endoperoxide H synthase-1 and -2 in NIH3T3 fibroblasts." Biochimica et Biophysica Acta. 1389. 57-66 (1998)
[Publications] Kitamura Y.: "Effect of IL-6 on tumor cell invasion of vascular endothelial monolayers." Surgery Today. 27(6). 534-541 (1997)
[Publications] Yamashita H.: "Growth/dufferentiation factor-5 induces angiogenesis in vivo." Experimantal Cell Research.235(1). 218-226 (1997)
[Publications] 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)
[Publications] Morita,I.: "Enhancement of membrane fluidity in cholesterol-poor endothelial cells pre-treated with simvastin." Endothelium. 5(2). 107-113 (1997)

1997 Fiscal Year Annual Research Report

アラキドン酸カスケードと病的動態

Principal Investigator

水柿 道直 東北大学, 医学部・附属病院, 教授 (60004595)

Research Products

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

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

[Publications] 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)

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

[Publications] 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)

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

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

[Publications] 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)

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

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

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

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

[Publications] 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)

[Publications] 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)

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

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

[Publications] 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)

[Publications] 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)

[Publications] 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).

[Publications] 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)

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

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

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

[Publications] 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)

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

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

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

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

[Publications] 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).

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

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

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

[Publications] 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)

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

水柿道直東北大学, 医学部・附属病院, 教授 (60004595)