Standardization and problems to measure prostanoids in the biological fluid.

测量生物体液中前列腺素的标准化和问题。

• 批准号: 03557039
• 负责人: ABE Keishi
• 金额: $ 10.56万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03557039/
• 关键词: Prostacyclin; Thromboxane A_2; 11-dehydro-thromboxane B_2; 2, 3-dinor-6-keto-prostaglandin F_1alpha; Hypertension; Toxemia of pregnancy; Inflammation; Measurement; トロンボキサン(TXA_2); プロスタサイクリン(PGI_2); 11-dehydro-TXB_2; 2,3-dinor-b-keto PGF_1α; 6,

The method to measure 11-dehydro-thromboxane A_2(TXA_2) in the biological fluid was established by the previous research project. In this following project we performed the clinical application of this method in various human diseases as well as animal models of diseases. Over productions of TXA_2 were suspected in patients with myocardial infarction, cerebral infarction, diabetes mellitus, toxemia of pregnancy and bronchial asthma who showed higher urinary excretions of 11-dehydro-TXB_2, Physiological and pathophysiological roles of TXA_2 were elucidated. Furthermore, various immunoaffinity column and HPLC separation method were developed for the clean up of eicosanoids from the biological fluids and the HPLC-laser system to detect the small amount of eicosanoids was also developed.On the other hand, EIA kit for 2, 3-dinor-6-keto-prostaglandin (PG)F_1alpha, the stable metabolite of prostacyclin (PGI_2) was developed and we shared in the work for the standardization and clinical applic … More ation of this method. The separation of 2, 3-dinor-6-keto-PGF_1alpha from biological fluid was needed by either immunoaffinity column using anti-6-keto-PGF_1alpha antibody or anti-2, 3-dinor-6-keto-PGF_1alpha antibody or by HPLC before the measurement by EIA.Urinary excretion of 2, 3-dinor-6-keto-PGF_1alpha in man was about 100-400 pg/mg creatinine measured either by EIA or by GC/SIM method. There were no sex related differences and urinary levels were affected mainly by the systemic production rate rather than the renal production rate. Thus the 2, 3-dinor-6-keto-PGF_1alpha seems suitable as the index of systemic production rate of PGI_2. This method was useful to measure the levels in the tissue culture media and in the biological fluid from inflammatory tissue. We also applied clinically to measure the urinary excretion of levels in patients with hypertension, diabetes mellitus and toxemia of pregnancy. The establishment of the measurement of major stable metabolites of TXA_2 and PGI_2, in turn, made it possible to clarify the production balance between TXA_2 and PGI_2 in various diseases. Surplus TXA_2 production than PGI_2 was found in patients with essential hypertension as well as diabetes mellitus.Thus, most important problems to measure TXA_2 and PGI_2 in the biological fluid was dissolved by these projects and these methods seems to be useful tools to clarify the pathphysiological roles of TXA_2 and PGI_2 in man. Less

本课题建立了生物体液中11-脱氢血栓烷A_2（TXA_2）的测定方法。在接下来的项目中，我们在各种人类疾病以及疾病的动物模型中进行了这种方法的临床应用。心肌梗塞、脑梗塞、糖尿病、妊娠毒血症、支气管哮喘等病人尿中11-dehydro-TXB_2增高者，可能存在TXA_2的过度产生。本文阐述了TXA_2的生理和病理生理作用。此外，还建立了多种免疫亲和柱和高效液相色谱分离方法，用于生物体液中类花生酸的净化，并建立了检测微量类花生酸的高效液相色谱-激光检测系统，同时还建立了2，3-二去甲-6-酮-前列腺素（PG）F_1 α酶免疫分析试剂盒，本文报道了前列环素（PGI_2）的稳定代谢产物的研制，并为PGI_2的标准化和临床应用做了工作。 ...更多信息 这个方法的作用。用EIA法或GC/SIM法测定人尿中2，3-dinor-6-keto-PGF_1 α的排泄量为100-400 pg/mg肌酐。无性别相关差异，尿液水平主要受全身生成率而非肾脏生成率的影响。因此，2，3-二去甲-6-酮-PGF_1 α似可作为PGI_2系统生成率的指标。该方法可用于测量组织培养基和来自炎症组织的生物流体中的水平。并应用于高血压、糖尿病及妊娠毒血症患者尿中的排泄量测定。TXA_2和PGI_2主要稳定代谢物测定方法的建立，又为阐明各种疾病中TXA_2和PGI_2的产生平衡提供了可能。在原发性高血压和糖尿病患者中发现TXA_2的产生多于PGI_2，从而解决了测定生物体液中TXA_2和PGI_2的最重要问题，这些方法似乎是阐明TXA_2和PGI_2在人体中的病理生理作用的有用工具。

项目成果

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K.Omata、N.G.Abraham 和 M.L.Schwartzman.：“自发性高血压大鼠肾细胞色素 P450 依赖性花生四烯酸代谢的不同形成。”

K.Omata, K.Abe, H.L.Sheu, K.Yoshida, E.Tsutsumi, K.Yoshinaga, N.G.Abraham and M.L.Schwartzman.: "Roles of renal cytochrome P450-dependent arachidonic acid metabolites in hypertension." Tohoku J.Exp.Med.166. 93-106 (1992)

K.Omata、K.Abe、H.L.Sheu、K.Yoshida、E.Tsutsumi、K.Yoshinaga、N.G.Abraham 和 M.L.Schwartzman.：“肾细胞色素 P450 依赖性花生四烯酸代谢物在高血压中的作用。”

K.Omata, E.Tsutsumi, H.L.Sheu, Y.Utsumi and K.Abe.: "Effect of aging on renal cytochrome P450-dependent arachidonic acid metabolism in Dahl rats." J.Lipid.Mediators.6. 369-373 (1993)

K.Omata、E.Tsutsumi、H.L.Sheu、Y.Utsumi 和 K.Abe.：“衰老对 Dahl 大鼠肾细胞色素 P450 依赖性花生四烯酸代谢的影响。”

K.Takeuchi, T.Abe, N.Takahashi and K.Abe.: "Molecular cloning and intrarenal localization of rat prostaglandin E2 receptor EP3 subtype." Biochem.Biophys.Res.Commun.194. 885-891 (1993)

K.Takeuchi、T.Abe、N.Takahashi 和 K.Abe.：“大鼠前列腺素 E2 受体 EP3 亚型的分子克隆和肾内定位。”

Masako Watanabe,Takanori Watanabe,Noriyhou Hirasawa,Suetsugu Mue,Shigeki Muramatsu,Yoko Matsushita,Hidekuni Takahagi,pierre Braquet collette Broquet,Lawrence Lcuine,KA〓: "Preparation of immunoaffinity mini-columns for the analysis of platlet activating fa

Masako Watanabe,Takanori Watanabe,Noriyhou Hirasawa,Suetsugu Mue,Shigeki Muramatsu,Yoko Matsushita,Hidekuni Takahagi,pierre Braquet collette Broquet,Lawrence Lcuine,KA〓：“用于分析血小板活化因子的免疫亲和微型柱的制备