Metabolic Markers of Carbonyl, Oxidative and Nitrosative Stress in the DCCT/EDIC

描述(由申请人提供)：高血糖水平已被确定为1型糖尿病微血管和大血管并发症进展的罪魁祸首和/或危险因素。我们对晚期糖基化终末产物(AGEs)的生化性质和临床意义的研究表明，在DCCT结束16年后，皮肤胶原蛋白AGEs是微血管和亚临床心血管并发症的强有力的长期预测因子。这种预测效果与糖化血红蛋白、年龄、糖尿病病程和其他危险因素无关。然而，除了固有的SKI荧光外，在工作台水平上测定皮肤中的特定年龄是不可行的。因此，如何将这些令人兴奋的发现转化为更可行的替代方法成为一项紧迫的任务。利用PAR声明的优势，我们建议调查一个实用的和临床相关的假设，即单独或一组基线或纵向测定血清和尿蛋白标记的羰基(AGEs)、氧化和亚硝化应激(“代谢应激标记”)，将使我们能够预测DCCT/EDIC试验中哪些1型糖尿病患者发生微血管和大血管并发症(主要终点)的长期风险增加。为了实现这一目标，组建了一个由凯斯西储大学DCCT/EDIC研究组的生物化学家和一名临床糖尿病专家调查员以及乔治华盛顿大学生物统计中心DCCT数据协调中心的统计学家组成的团队。数据将使用带有同位素稀释的强大的液-色质谱方法产生，该方法准确地测定9个标志物，即2个糖基化产物(果糖-赖氨酸和葡萄糖)，2个糖氧化产物(羧甲基赖氨酸(CML)和羧乙基-赖氨酸(CEL)，2个甲醛产物(乙二醛和甲基乙醛氢咪唑酮)，2个金属催化氧化产物(大蒜氨酸和2-氨基己二酸)和一个氧化产物(蛋氨酸亚磺酸)。用荧光高效液相色谱法测定戊二胺和亚硝化产物3-硝基酪氨酸。将对统计模型进行测试，以确定这些血清和尿液标记物与皮肤年龄的关系，如何促进并发症的长期风险，以及对已知风险因素的调整是否削弱了这些标记物的预测能力。这项研究的成功结果可能会导致实施“代谢应激小组”，作为HbA1c的辅助工具，用于在临床环境中评估糖尿病并发症的代谢危险因素。

DESCRIPTION (provided by applicant): Elevated glucose levels have been identified as culprits and/or risk factors for the progression of microvascular and macrovascular complications in Type 1 diabetes. Our studies on the biochemical nature and clinical significance of advanced glycation endproducts (AGEs) determined near DCCT closeout demonstrated that skin collagen AGEs are robust long-term predictors of microvascular and subclinical cardiovascular complications 16 years after the end of DCCT. This predicting effect is, independent of HbA1c, age, diabetes duration, and other risk factors. However, the determination of specific AGEs in skin is not feasible at the bench level, except for intrinsic ski fluorescence. Therefore, how to translate these exciting findings to a more feasible alternative approach becomes a pressing task. Taking advantage of the PAR announcement, we propose to investigate a pragmatic and clinically relevant hypothesis that determination of serum and urine protein markers of carbonyl (AGEs), oxidative and nitrosative stress ("metabolic stress markers"), individually or a set, at baseline or longitudinally, will allow us to predict which individuals with Type 1 diabetes in the DCCT/EDIC trial are at long-term increased risk of developing micro- and macrovascular vascular complications (primary endpoints). To achieve this goal, a team consisting of biochemists and a clinical diabetologist investigator from the DCCT/EDIC Research Group at Case Western Reserve University and statisticians at theDCCT Data Coordinating at the Biostatistics Center of the George Washington University has been assembled. Data will be generated using a powerful liquid chromatography mass spectrometry method with isotope dilution that accurately determines 9 markers, i.e. 2 glycation products (fructose-lysine and glucose pane), 2 glycoxidation products (carboxymethyl-lyinse (CML) and carboxyethyl-lysine (CEL), 2 oxoaldehyde products (glyoxal and methylglyoxal hydroimidazolones), 2 products of metal catalyzed oxidation (allysine and 2-amino adipic acid) and one oxidation product (methionine sulfoxide). Pentosidine and the nitrosylation product 3-nitrotyrosine will be determined by fluorescence HPLC. Statistical models will be tested to determine how these serum and urine markers relate to skin AGEs, contribute to long-term risk of complications, and whether adjustment for known risk factors attenuates the predicting power of these markers. A successful outcome of this research may lead to the implementation of a "metabolic stress panel" as adjunct to HbA1c for the evaluation of metabolic risk factors for diabetic complications in the clinical setting.