Systematic Quantitation of Glycoxidative Stress in Biological Systems

葡萄糖是细胞中最丰富的单糖。它具有化学反应性，可以与不同类别的生物分子（例如氨基酸和蛋白质）发生不需要的反应，影响它们的理化性质和潜在的生物功能。这种反应被称为非酶糖化，通常发生速度较慢，是生物体衰老过程的主要组成部分。多种生理并发症，例如糖尿病、白内障、神经退行性疾病和心血管疾病都与非酶糖化有关。 先前的研究表明，该反应在氧化应激条件下会加速。生物金属，例如铁和铜，可以诱导氧化并促进氧化反应。因此，一些研究假设选择性降低内源性金属水平可能对非酶糖化具有控制作用。人们发现，几类天然化合物与过量的促氧化金属发生反应，对人体中不需要的糖化具有抑制作用。然而，其根本机制尚未完全了解。该研究计划的总体目标是开发定量研究生物系统中非酶糖化的方法。通过这个项目，我们的目标是识别新的生物标志物，帮助我们更好地了解系统水平的糖化反应。在目前的资助期内，将开发一个实验模型，用于体外非酶糖化研究，重点关注低分子量代谢物（目标 1）。将创建并测试分析方法，以对生物样品中主要糖化产物进行全面分析和有针对性的定量（目标 2）。随后将开发并验证多重测定，以监测细胞培养模型中存在和不存在过量促氧化剂金属的情况下非酶糖化的速率和程度（目标 3）。这些成果对于饮食学、衰老研究、诊断以及糖氧化应激相关疾病及其分子发病机制的理解具有重要价值。该研究计划采用了新的方法和假设，并为培训本科生和研究生提供了机会。将通过与加拿大保健食品技术和分子诊断行业领导者的合作，制定将本文的发现推向市场的策略。该研究计划将对我们对非酶糖化及其众多生理后果的了解以及加拿大在自然科学领域技术开发的全球领导地位产生重大影响。

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.