Systematic Quantitation of Glycoxidative Stress in Biological Systems

葡萄糖是细胞中含量最丰富的单糖。它具有化学反应活性，可以与不同类别的生物分子，如氨基酸和蛋白质发生不必要的反应，影响它们的物理化学性质，并可能影响它们的生物功能。这种反应被称为非酶糖基化，通常以缓慢的速度发生，是生物体衰老过程的主要组成部分。许多生理并发症，如糖尿病、白内障、神经退行性疾病和心血管疾病，都与非酶糖基化有关。以往的研究表明，在氧化应激条件下，这种反应会加速。生物金属，如铁和铜，可以诱导氧化并促进氧化反应。因此，一些研究推测，选择性降低内源性金属水平可能对非酶糖化有控制作用。研究发现，几类天然化合物与过量的促氧化剂金属反应，对人类不希望发生的糖基化具有抑制作用；然而，其潜在的机制尚未完全清楚。这项研究计划的首要目标是开发生物系统中非酶糖基化的定量研究方法。通过这项计划，我们将致力于识别新的生物标志物，帮助我们在系统水平上更好地了解糖基化反应。在目前的资助期间，将开发一个实验模型来研究体外非酶糖化作用，重点是低分子代谢物(目标1)。将建立和测试生物样品中主要糖基化产物的全面剖析和有针对性的定量的分析方法(目标2)。随后将开发和验证一种多重分析方法，以监测细胞培养模型中存在和不存在过量促氧化金属的情况下非酶糖化的速度和程度(目标3)。这些成果将对营养学、衰老研究、诊断和了解糖氧化应激相关疾病及其分子发病机制具有重要意义。这项研究计划结合了新的方法和假设，并为培养本科生和研究生提供了一个机会。将通过与加拿大健康食品技术和分子诊断领域的行业领先者合作，制定将这里的发现推向市场的战略。这项研究计划将对我们对非酶糖基化及其众多生理后果的知识产生重大影响，并对加拿大在自然科学领域的技术发展方面的全球领先地位产生重大影响。

Glucose is the most abundant monosaccharide in the cell. It is chemically reactive and can undergo an undesired reaction with different classes of biomolecules, such as amino acids and proteins, affecting their physicochemical properties and potentially their biological functions. Known as non-enzymatic glycation, this reaction generally occurs at a slow rate and is a major component of the aging process in living organisms. A broad range of physiological complications, such as diabetes, cataracts, and neurodegenerative and cardiovascular diseases has been linked to non--enzymatic glycation. Previous studies have shown that the reaction accelerates under oxidative stress conditions. Biometals, such as iron and copper, can induce oxidation and facilitate oxidative reactions. Some studies have, therefore, postulated that selective decreasing of endogenous metal levels may have a controlling effect on non-enzymatic glycation. Several classes of natural compounds that react with the excess amounts of prooxidant metals were found to have an inhibitory effect on undesired glycation in humans; however, the underlying mechanisms are not fully understood. The overarching goal of this research program is to develop methods for quantitative study of non-enzymatic glycation in biological systems. Through this program, we will aim to identify new biomarkers that can help us better understand the glycation reaction at a systemic level. In the present funding period, an experimental model will be developed for the study of non-enzymatic glycation in vitro with a focus on low molecular weight metabolites (objective 1). Analytical methods will be created and tested for comprehensive profiling and targeted quantitation of major glycation products in biological samples (objective 2). A multiplex assay will be subsequently developed and validated to monitor the rate and extent of non-enzymatic glycation in the presence and absence of excess prooxidant metals in a cell culture model (objective 3). These achievements will be valuable to dietetics, aging research, diagnosis, and understanding of glycoxidative stress-related disorders and their molecular pathogenesis. This research program incorporates new methods and hypotheses and presents an opportunity for training undergraduate and graduate students. Strategies to bring discoveries made herein to market will be developed through partnership with Canadian industry leaders in health food technologies and molecular diagnostics. This research program will have a significant impact on our knowledge of non-enzymatic glycation, its numerous physiological consequences, and on Canada's global leadership in technology development in the field of natural sciences.