Evaluation of in vivo enzymatic activity of cortisol metabolism associated with hypertension

与高血压相关的皮质醇代谢的体内酶活性评估

• 批准号: 12672225
• 负责人: KASUYA Yasuji
• 金额: $ 1.98万
• 依托单位: Tokyo University of Pharmacy and life Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12672225/
• 关键词: cortisol; cortisone; stable isotope methodology; 11β-HSD activity; hypertension; GC-MS-SIM; 13C_4-and 2H_1-label; pharmacokinetic analysis; 安定同位体標識; 11β-HSD2酵素活性評価; GC-MS分析

This study is concerned with oral administrations of [1,2,4,19-13C_4] cortisol (cortisol-13C_4) and [1,2,4,19-13C_4,11α-^2H] cortisol (cortisol-13C_4,^2H_1) to a human subject to reliably evaluate the individual activities of two isozymes of 11β-HSD. The use of a GC-MS method allowed the simultaneous measurement of the plasma concentrations of cortisol-13C_4, ^2H_1, cortisone-13C_4, and cortisol-13C_4 together with endogenous cortisol and cortisone. The lossof 11α-^2H during the conversion of cortisol-13C_4, ^2H_1 to cortisone-13C_4 by 11β-HSD2 and the regenerated cortisol-13C_4 from cortisone-13C_4 by 11β-HSD1 provided a direct and accurate means of distinguishing the activities of the two isozymes. The kinetic analysis associated with the metabolism of orally administered cortisol-13C_4 and cortisol-13C_4, ^2H1 was of great importance in assessing the 11β-HSD activities.This stable isotope approach offered unique advantages since it allowed to measure stable isotopically labeled cortisol and its labeled metabolite cortisone simultaneously with endogenous (unlabeled) cortisol and cortisone. Very important was the potential use of stable isotopically labeled cortisol in man as a tracer for the native cortisol to recognize the conversion of cortisol to cortisone in the kidney. Our results from the stable isotope methodology strongly suggest that cortisol-13C_4, ^2H_1 used in this study served as an appropriate tracer for elucidating the kinetics of the interconversion of cortisol to cortisone in man.

本研究对人体口服[1，2，4，19-13C_4]皮质醇(Cortisol-13C_4)和[1，2，4，19-13C_4，11α-^2H]皮质醇(Cortisol-13C_4，^2H_1)进行了研究，以可靠地评价11β-HSD两种同工酶的个体活性。用GC-MS法可同时测定血浆中皮质醇-13C_4、皮质醇-13C_4、皮质醇-13C_4、皮质醇和皮质醇的浓度。11α-HSD2将皮质醇-13C_4、^2H_1转化为皮质醇-13C_4的过程中损失的11β-2H和11β-HSD1从皮质醇-13C_4中再生的皮质醇-13C_4为区分这两种同工酶的活性提供了一种直接、准确的手段。与口服皮质醇-13C4和皮质醇-13C4，2H1代谢相关的动力学分析在评估11种β-HSD活性方面具有重要意义。这种稳定同位素方法具有独特的优势，因为它允许同时测量稳定的同位素标记的皮质醇及其标记的代谢物皮质醇和内源性(未标记的)皮质醇和皮质松。非常重要的是在人体中使用稳定的同位素标记的皮质醇作为天然皮质醇的示踪剂来识别肾脏中皮质醇转化为皮质醇的可能性。我们的稳定同位素方法学结果有力地表明，本研究中使用的皮质醇-13C_4，^2H_1可作为阐明人体皮质醇相互转化为皮质醇的动力学的合适示踪剂。

项目成果

Kasuya Y, Yokokawa A, Takayama S, Shibasaki H, and Furuta T.: "Evaluation of 11β-HSD activities in vivo following oral administration of cortisol-13C_4, 2H_1 to a human subject"Steroid. 68. 167-176 (2003)

Kasuya Y、Yokokawa A、Takayama S、Shibasaki H 和 Furuta T.：“对人类受试者口服皮质醇-13C_4、2H_1 后体内 11β-HSD 活性的评估”类固醇。

Furuta T, Eguchi N, Yokokawa A, Shibasaki H, and Kasuya Y.: "Synthesis of multi-labeled cortisols and cortisones with 2H and 13C for study of cortisol metabolism in humans."Steroids. 65. 180-189 (2000)

Furuta T、Eguchi N、Yokokawa A、Shibasaki H 和 Kasuya Y.：“多标记皮质醇和具有 2H 和 13C 的可的松的合成，用于研究人类皮质醇代谢。”类固醇。

Furuta T, Efuchi N, Shibasaki H, and Kasuya Y.: "Simultaneous determination of endogenous and 13C-labelled cortisols and cortisones in human plasma by stable isotope dilution mass spectrometry."J. Chromatogr. B, Biomed. Sci.. 738. 119-127 (2000)

Furuta T、Efuchi N、Shibasaki H 和 Kasuya Y.：“通过稳定同位素稀释质谱法同时测定人血浆中的内源性和 13C 标记的皮质醇和可的松。”J.

Kasuya Y, Yokokawa A, Furuta T, Shibasaki H: "Measurement of cortisol secretion rate by stable isotope methodology following oral administration of ^<13>C-labeled cortisol to a human subject"Steroids. 67・9. 783-788 (2002)

Kasuya Y、Yokokawa A、Furuta T、Shibasaki H：“对人类受试者口服 13 C 标记的皮质醇后通过稳定同位素方法测量皮质醇分泌率”Steroids 67·9。 ）

Kasuya Y, Yokokawa A.Furuta T, Shibasaki H: "Measurement of cortisol secretion rate by stable isotope methodology following oral administration of ^<13>C-labeled cortisol to a human subject"Steroids. 67・9. 783-788 (2002)

Kasuya Y、Yokokawa A.Furuta T、Shibasaki H：“对人类受试者口服^ 13 C标记的皮质醇后通过稳定同位素方法测量皮质醇分泌率”Steroids 67・9。 ）