Dirsch V M, Stuppner H, Vollmar A M
Institut für Pharmakologie, Toxikologie und Pharmazie, Ludwig-Maximilians-Universität München, Germany.
Planta Med. 1998 Jun;64(5):423-6. doi: 10.1055/s-2006-957473.
In the field of inflammation research the inducible nitric oxide synthase (iNOS) became an important pharmacological target, since overproduction of nitric oxide (NO) after induction of this enzyme seems to be associated with numerous pathological conditions. NO released from cells can be detected and quantified photometrically as its stable product nitrite by a simple colorimetric reaction (Griess reaction). The aim of our study was to investigate whether this method might be suitable for the bio-guided fractionation of anti-inflammatory plant extracts. For this purpose we assayed extracts as well as fractions of the roots of Curcuma zanthorrhiza Roxb, which contain the known iNOS inhibitor curcumin, and compared the obtained activity with their curcumin content. Furthermore, leaf extracts of Betula pendula Roth, to which defined amounts of curcumin were added, were examined to clarify the question whether chlorophyll might interfere with the test system. The presented results suggest that the Griess assay is indeed suitable to guide fractionation of plant extracts in order to isolate highly active compounds. Factors, however, which might restrict the broad application of this assay are the limited selection of solvents which do not interfere with the system and high contents of chlorophyll in plant extracts.
在炎症研究领域，诱导型一氧化氮合酶（iNOS）已成为一个重要的药理学靶点，因为该酶诱导后一氧化氮（NO）的过量产生似乎与众多病理状况相关。细胞释放的NO可以通过简单的比色反应（格里斯反应），作为其稳定产物亚硝酸盐进行光度检测和定量。我们研究的目的是调查这种方法是否适用于抗炎植物提取物的生物导向分级分离。为此，我们检测了含有已知iNOS抑制剂姜黄素的郁金的根提取物及其馏分，并将获得的活性与其姜黄素含量进行比较。此外，还检测了添加了一定量姜黄素的欧洲白桦叶提取物，以阐明叶绿素是否会干扰测试系统这一问题。给出的结果表明，格里斯测定法确实适用于指导植物提取物的分级分离，以分离高活性化合物。然而，可能限制该测定法广泛应用的因素是不干扰该系统的溶剂选择有限以及植物提取物中叶绿素含量高。
