Vaya Jacob
Laboratory of Natural Medicinal Compounds, Migal-Galilee Technology Center, Kiryat Shmona, Israel.
Methods Mol Biol. 2008;477:3-13. doi: 10.1007/978-1-60327-517-0_1.
Oxidative stress (OS) is linked to the development of human diseases. Early identification of OS-associated diseases is essential in the control of their progression and treatment. Efforts have been undertaken to identify reliable endogenous markers, which correlate with the progression of a disease in an organ undergoing OS. An ideal biomarker must be validated, utilize noninvasive sampling, and have a simple, specific and highly sensitive detection method. Among the currently used markers assessing OS, are those that are nonspecific (peroxide value [PV], conjugated dienes [CD], thiobarbitoric acid reactive substances [TBARS]), and others that measure end-products of oxidized degradation biomolecules (isoprostanes, oxysterols, keto-proteins, 8-oxodeoxyguanosine), whose accumulation is not necessarily correlated with augmented OS. The search for a more reliable marker necessitates new approaches to fulfill such requirements and overcome many of the obstacles associated with the current markers. We suggest a new strategy of using designed exogenous novel reporters, constructed from endogenous subunits, that are sensitive to reactive oxygen and nitrogen species (ROS/RNS) and commonly known to react with them, forming specific oxidized products. These subunits are tyrosine (representing proteins), bonded covalently to linoleic acid (representing polyunsaturated fatty acids) forming an amide bond, which can be further connected through an ester bond to a third unit, either to cholesterol (representing sterols) or to 2'-deoxyguanosine (representing DNA). Oxidation of the designed probe can outline, in real time, the formation of oxidation products and distinguish them from intrinsic biomolecules, provide information about the relative subunit susceptibilities to a specific oxidant challenge, and allow for the assessment of the utility of intervention, such as antioxidant supplementation. By utilizing such markers, it may be possible to correlate between the damaged fingerprints of the marker and the specific pathological conditions. The above markers were tested to characterize OS in in vitro and in in vivo experiments, such as in those carried out in human fluids (blood, serum, saliva), tissues (brain or muscle homogenates), and cells (macrophages, astrocytes, neurons), pertaining to OS-associated diseases, such as atherosclerosis, diabetes, and Alzheimer's disease.
氧化应激（OS）与人类疾病的发展相关。早期识别与OS相关的疾病对于控制其进展和治疗至关重要。人们一直在努力寻找可靠的内源性标志物，这些标志物与经历OS的器官中疾病的进展相关。理想的生物标志物必须经过验证，采用非侵入性采样，并且具有简单、特异且高度灵敏的检测方法。在目前用于评估OS的标志物中，有些是非特异性的（过氧化值[PV]、共轭二烯[CD]、硫代巴比妥酸反应性物质[TBARS]），还有一些是测量氧化降解生物分子的终产物（异前列腺素、氧化甾醇、酮蛋白、8-氧代脱氧鸟苷），其积累不一定与增强的OS相关。寻找更可靠的标志物需要新的方法来满足这些要求并克服与当前标志物相关的许多障碍。我们提出了一种新策略，即使用由内源性亚基构建的设计外源性新型报告分子，这些报告分子对活性氧和氮物种（ROS/RNS）敏感，并且通常已知会与它们反应，形成特定的氧化产物。这些亚基是酪氨酸（代表蛋白质），通过酰胺键与亚油酸（代表多不饱和脂肪酸）共价结合，然后可以通过酯键进一步与第三个单元连接，该单元可以是胆固醇（代表甾醇）或2'-脱氧鸟苷（代表DNA）。设计探针的氧化可以实时勾勒出氧化产物的形成，并将它们与内源性生物分子区分开来，提供有关亚基对特定氧化剂挑战的相对敏感性的信息，并允许评估干预措施（如补充抗氧化剂）的效用。通过使用此类标志物，有可能将标志物的受损指纹与特定病理状况相关联。上述标志物已在体外和体内实验中进行测试，以表征OS，例如在与OS相关疾病（如动脉粥样硬化、糖尿病和阿尔茨海默病）相关的人体体液（血液、血清、唾液）、组织（脑或肌肉匀浆）和细胞（巨噬细胞、星形胶质细胞、神经元）中进行的实验。
