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。

基本信息

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

项目摘要

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

之前的研究项目建立了测定生物体液中11-脱氢血栓烷A_2(TXA_2)的方法，在本项目中我们将该方法在多种人类疾病以及动物疾病模型中进行了临床应用。患有心肌梗塞、脑梗塞、糖尿病、妊娠毒血症和支气管哮喘的患者怀疑会产生TXA_2，这些患者的尿排泄量较高阐明了 11-脱氢-TXB_2、TXA_2 的生理和病理生理作用。此外，还开发了多种免疫亲和柱和 HPLC 分离方法来净化生物体液中的类二十烷酸，并开发了 HPLC-激光系统来检测少量的类二十烷酸。另一方面，还开发了2, 3-dinor-6-keto-prostaglandin的EIA试剂盒开发了前列环素 (PGI_2) 的稳定代谢物 (PG)F_1alpha，并参与了该方法的标准化和临床应用工作。从生物中分离 2, 3-dinor-6-keto-PGF_1alpha。使用抗 6-酮-PGF_1α 抗体或抗 2 抗体的免疫亲和柱需要液体， 3-dinor-6-keto-PGF_1alpha 抗体或在 EIA 测量之前通过 HPLC。通过 EIA 或通过GC/SIM 方法没有性别相关的差异，并且尿水平主要受全身生成率而非肾脏生成率的影响。 2, 3-dinor-6-keto-PGF_1alpha 似乎适合作为 PGI_2 的全身生成率的指标。该方法可用于测量组织培养基和炎症组织的生物液中的水平。我们也应用于临床。测定高血压、糖尿病、妊娠毒血症患者尿排泄水平。建立TXA_2和PGI_2主要稳定代谢物测定方法。反过来，使得阐明各种疾病中 TXA_2 和 PGI_2 之间的产生平衡成为可能，在原发性高血压和糖尿病患者中发现 TXA_2 产生量比 PGI_2 多。因此，测量生物体液中 TXA_2 和 PGI_2 是最重要的问题。这些项目解决了这些问题，这些方法似乎是阐明 TXA_2 和 PGI_2 在人类中的病理生理作用的有用工具。