Koss-Mikołajczyk Izabela, Baranowska Monika, Namieśnik Jacek, Bartoszek Agnieszka
Katedra Chemii Analitycznej, Wydział Chemiczny, Politechnika Gdańska.
Katedra Chemii, Technologii i Biotechnologii Żywności, Wydział Chemiczny, Politechnika Gdańska.
Postepy Hig Med Dosw (Online). 2017 Jul 30;71(0):602-617. doi: 10.5604/01.3001.0010.3841.
As soon as the role of Reactive Oxygen Species (ROS) in so-called civilization diseases, which include non-infectious chronic diseases such as cancer, diabetes or high blood pressure has been discovered, and the possibility of employing antioxidants as a remedy for these diseases have been proposed, scientists developed a broad spectrum of methods to determine antioxidant activity of pure chemicals and plant extracts, as well as dietary supplements. Most of these methods are based on simple redox reactions between antioxidant and ROS (for example ABTS, DPPH, or FRAP tests). However, chemical methods of assessing antioxidant activity are rarely biologically relevant. They do not mirror the real effect of antioxidants in living organisms, because they are used in non-physiological conditions of temperature and pH; neither they take metabolism nor intracellular transport under consideration. The perfect model for assessment of antioxidant activity in living organisms would be human or animal model, but such determinations are very complicated and often ambiguous. The current best alternative to chemical and human tests are assays employing cell culture models being less expensive than human tests, yet still reflecting biological systems more convincingly than chemical assays. Cellular antioxidant assays are performed under physiological pH and temperature, but most importantly, they take metabolism and intracellular transport under consideration. In this review, we present cellular tests used to determine antioxidant activity that are based on luminescence and fluorescence methods.
一旦发现活性氧（ROS）在所谓的文明病（包括癌症、糖尿病或高血压等非传染性慢性病）中的作用，并且提出了使用抗氧化剂治疗这些疾病的可能性，科学家们就开发了一系列广泛的方法来测定纯化学品、植物提取物以及膳食补充剂的抗氧化活性。这些方法大多基于抗氧化剂与ROS之间的简单氧化还原反应（例如ABTS、DPPH或FRAP测试）。然而，评估抗氧化活性的化学方法在生物学上很少具有相关性。它们无法反映抗氧化剂在活生物体中的实际作用，因为它们是在非生理温度和pH条件下使用的；它们也没有考虑代谢和细胞内运输。评估活生物体中抗氧化活性的完美模型应该是人类或动物模型，但这样的测定非常复杂且往往含糊不清。目前化学测试和人体测试的最佳替代方法是采用细胞培养模型的测定方法，这种方法比人体测试成本更低，但仍比化学测定更令人信服地反映生物系统。细胞抗氧化测定是在生理pH和温度下进行的，但最重要的是，它们考虑了代谢和细胞内运输。在这篇综述中，我们介绍了基于发光和荧光方法用于测定抗氧化活性的细胞测试。
