大鼠肾上腺儿茶酚 - O - 甲基转移酶活性测定方法：自发性高血压大鼠与Wistar - Kyoto大鼠的比较

An assay for determination of rat adrenal catechol-O-methyltransferase activity: comparison of spontaneously hypertensive rats and Wistar-Kyoto rats.

作者信息

Tsunoda Makoto, Imai Kazuhiro

机构信息

Laboratory of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

出版信息

Anal Bioanal Chem. 2004 Dec;380(7-8):887-90. doi: 10.1007/s00216-004-2884-7. Epub 2004 Nov 13.

DOI:10.1007/s00216-004-2884-7

PMID:15551073

Abstract

A method has been developed for measurement of catechol-O-methyltransferase (COMT) activity in the rat adrenal gland. Epinephrine, synthesized in the adrenal gland, was used as substrate, and its enzymatic product, metanephrine, was quantified by high-performance liquid chromatography (HPLC) with fluorescence detection. The method has sufficient precision and accuracy. Soluble (S) and membrane-bound (MB) COMT activity in Wistar-Kyoto (WKY) rats was 20.7 +/- 3.5 and 18.6 +/- 3.4 pmol min(-1) mg(-1) protein (n = 5), respectively. To clarify the role of adrenal COMT in blood-pressure regulation, S and MB COMT activity in spontaneously hypertensive rats were determined. Respective activity was 18.6 +/- 3.4 and 17.0 +/- 1.1 pmol min(-1) mg(-1) (n = 5), which is similar to that in WKY rats. This finding suggests that COMT in the adrenal gland might not be related to blood pressure regulation.

摘要

已开发出一种测量大鼠肾上腺中儿茶酚-O-甲基转移酶（COMT）活性的方法。肾上腺中合成的肾上腺素用作底物，其酶促产物间甲肾上腺素通过高效液相色谱（HPLC）荧光检测进行定量。该方法具有足够的精密度和准确度。Wistar-Kyoto（WKY）大鼠的可溶性（S）和膜结合（MB）COMT活性分别为20.7±3.5和18.6±3.4 pmol min⁻¹ mg⁻¹蛋白质（n = 5）。为阐明肾上腺COMT在血压调节中的作用，测定了自发性高血压大鼠的S和MB COMT活性。各自的活性为18.6±3.4和17.0±1.1 pmol min⁻¹ mg⁻¹（n = 5），与WKY大鼠相似。这一发现表明肾上腺中的COMT可能与血压调节无关。