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[利用蛋白质印迹法对生药和汉方药进行科学评价及其在生物代谢研究中的应用]

[Scientific Evaluation of Crude Drugs and Kampo Medicines Using the Eastern Blotting Method and Its Application to Biological Metabolic Studies].

作者信息

Morinaga Osamu

机构信息

Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Nagasaki International University.

出版信息

Yakugaku Zasshi. 2018;138(6):743-750. doi: 10.1248/yakushi.17-00204.

Abstract

The scientific evaluation of crude drugs and kampo medicines (KMs) was demonstrated using the eastern blotting method with monoclonal antibodies (MAbs) against bioactive natural compounds. Scutellariae radix is one of the most important crude drugs used in KMs. Part of its pharmaceutical properties is due to the flavone glycoside baicalin (BI). A quantitative analysis method based on eastern blotting was developed for BI using an anti-BI MAb. A rapid, simple, sensitive, specific analytical system was subsequently established for BI with the eastern blotting technique using dot-blot and chemiluminescent methods. This system was useful as a high-throughput analytical method for the determination of BI in KMs as well as HPLC and enzyme-linked immunosorbent assay systems. Furthermore, an eastern blotting method was applied to the biological metabolic study of glycyrrhizic acid (GL), the major active constituent of licorice, for investigation of metabolites of GL such as 3-monoglucuronyl-glycyrrhetinic acid (3MGA) because licorice causes pseudoaldosteronism as a side effect. This approach may make it possible to determine the pathogenic agents of licorice-induced pseudoaldosteronism.

摘要

利用针对生物活性天然化合物的单克隆抗体(MAb)的蛋白质免疫印迹法,对生药和汉方药(KM)进行了科学评估。黄芩是KM中最重要的生药之一。其部分药理特性归因于黄酮苷黄芩苷(BI)。使用抗BI单克隆抗体建立了基于蛋白质免疫印迹法的BI定量分析方法。随后,采用斑点印迹和化学发光方法,利用蛋白质免疫印迹技术为BI建立了一种快速、简单、灵敏、特异的分析系统。该系统作为一种高通量分析方法,可用于测定KM中的BI,也可用于高效液相色谱(HPLC)和酶联免疫吸附测定系统。此外,蛋白质免疫印迹法被应用于甘草主要活性成分甘草酸(GL)的生物代谢研究,以研究GL的代谢产物,如3-单葡萄糖醛酸基甘草次酸(3MGA),因为甘草会引起假性醛固酮增多症作为副作用。这种方法可能有助于确定甘草诱导的假性醛固酮增多症的致病因子。

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