In Vivo Regulation of Protein Turnover by Hormones

激素对蛋白质周转的体内调节

• 批准号: 8512150
• 负责人: K Sreekumaran Nair
• 金额: $ 14.37万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-19 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8512150
• 关键词: 8-Oxo-2' -Deoxyguanosine; ATP Synthesis Pathway; Address; Adverse effects; Aerobic Exercise; Age; American; Antioxidants; Asian Indian; Attenuated; Biological; Buffers; Caloric Restriction; Cells; Central obesity; Characteristics; Chemicals; Coupling; DNA; DNA biosynthesis; Data; Deoxyguanosine; Development; Diabetes Mellitus; Diet; Elderly; Energy Metabolism; European; Exercise; Exhibits; Fat-Restricted Diet; Fatty acid glycerol esters; Functional disorder; Funding; Gel; Genes; Health; Hormones; Immigrant; Individual; Infusion procedures; Insulin; Insulin Resistance; Intervention; Isotopes; Laboratories; Lipids; Measures; Metabolic Diseases; Methodology; Methods; Mitochondria; Mitochondrial DNA; Muscle; Muscle Proteins; Non-Insulin-Dependent Diabetes Mellitus; Nucleic Acids; Obesity; Oxidative Stress; Participant; Pharmaceutical Preparations; Phenylalanine; Physical activity; Placebos; Plasma; Plasma Proteins; Play; Population; Post-Translational Protein Processing; Predisposition; Prevalence; Process; Production; Protein Biosynthesis; Protein Isoforms; Proteins; Proteome; Reactive Oxygen Species; Regulation; Relative (related person); Reporting; Risk; Role; Seminal; Skeletal Muscle; Societies; Spottings; System; Transcript; Two-Dimensional Gel Electrophoresis; base; deamidation; density; improved; in vivo; insight; insulin sensitivity; insulin sensitizing drugs; novel; novel strategies; oxidation; oxidative DNA damage; oxidative damage; protein degradation; protein metabolism; public health relevance; research study; response; sex; therapy design

DESCRIPTION (provided by applicant): We will first determine whether people with high muscle mitochondrial capacity produce higher amount of reactive oxygen species (ROS) on consuming high fat diet and thus exhibit elevated cellular oxidative damage. We previously found that Asian Indian immigrants have high mitochondrial capacity in spite of severe insulin resistance. Somalians are another new immigrant population with rapidly increasing prevalence of diabetes. Both of these groups traditionally consume low caloric density diets, and we hypothesize that when these groups are exposed to high-calorie Western diets, they exhibit increased oxidative stress, oxidative damage, and insulin resistance. We will compare Asian Indians, Somalians, and NE Americans who are matched for age, BMI, and sex. We will measure ROS production in skeletal muscle following high fat meal. We will compare the oxidative damage to proteins, DNA, and lipids in these three populations following 2 weeks of high fat diet in comparison with low fat diet. We will determine if elevated levels of oxidative damage in these populations is accompanied by high mitochondrial capacity, higher ROS-emitting potential, and lower insulin sensitivity than NE. Secondly the proposal will apply a novel method developed in our laboratory to determine whether abdominally obese people with severe insulin resistance, high insulin levels, and oxidative stress accumulate old and damaged muscle and plasma proteins that may cause deleterious functional consequences. We will determine whether decreasing circulating insulin and oxidative stress by enhancing insulin sensitivity decreases the accumulation of old and damaged protein in muscle and plasma. We hypothesize that three different modes of enhancing insulin sensitivity act by different mechanisms to decrease the accumulation of old and damaged proteins and DNA oxidation. We expect caloric restriction (CR) to reduce oxidative stress by reducing ROS production and thus decrease oxidative damage to proteins and DNA. In contrast aerobic exercise increases ROS production but stimulates a robust antioxidant defense system. Furthermore, unlike CR we expect exercise to increase the synthesis of nascent proteins and turnover of old, damaged proteins. Insulin sensitizers reduce oxidative stress and enhance clearance of old and damaged protein. We will study abdominally obese people and compare them with lean participants to determine whether these insulin resistant people accumulate more damaged proteins and DNA in comparison with lean insulin sensitive people. We will then determine whether 16 weeks of aerobic exercise, caloric restriction and insulin sensitizers versus placebo will attenuate oxidative damage to proteins and DNA and improve insulin sensitivity. The proposed studies will be performed utilizing the state-of-the-art methods many of which were developed in our laboratory during the previous funding period. We expect the results from these studies to provide seminal insights into the underlying mechanism of insulin resistance and type 2 diabetes, in addition to demonstrating mechanisms by which a functional proteome is maintained in vivo. PUBLIC HEALTH RELEVANCE: The proposed experiments investigate the potential novel underlying mechanisms of rapidly expanding prevalence of diabetes and related health problems in the society. First specific aim is to determine whether more efficient mitochondria, the powerhouse of cell that produce chemical energy, causes adverse effects on DNA and protein when transitioning from a diet of low calorie content to that of high energy content. The second and third aims are to measure the accumulation of damaged and old proteins in muscle and plasma, using a novel methodology in obese and insulin resistant people and to determine how this potentially deleterious process is altered by aerobic exercise, caloric restriction, and medication that enhance insulin sensitivity and reduce high insulin levels.

描述(申请人提供)：我们将首先确定肌肉线粒体能力高的人在摄入高脂肪饮食时是否会产生更多的活性氧物种(ROS)，从而表现出更高的细胞氧化损伤。我们此前发现，尽管存在严重的胰岛素抵抗，亚裔印度移民仍具有很高的线粒体能力。索马里人是糖尿病患病率迅速上升的另一个新移民群体。这两组人传统上都食用低热量密度的饮食，我们假设，当这些组接触到高卡路里的西方饮食时，他们表现出更高的氧化应激、氧化损伤和胰岛素抵抗。我们将比较年龄、体重指数和性别相匹配的亚裔印度人、索马里人和东北美洲人。我们将测量高脂肪饮食后骨骼肌中ROS的产生。我们将比较两周高脂肪饮食和低脂肪饮食对这三个人群蛋白质、DNA和脂质的氧化损伤。我们将确定这些人群中氧化损伤水平的升高是否伴随着比NE更高的线粒体容量、更高的ROS释放潜力和更低的胰岛素敏感性。其次，该提案将应用我们实验室开发的一种新方法来确定具有严重胰岛素抵抗、高胰岛素水平和氧化应激的腹型肥胖者是否积累了可能导致有害功能后果的老化和受损的肌肉和血浆蛋白质。我们将确定是否通过增强胰岛素敏感性来减少循环中的胰岛素和氧化应激，从而减少肌肉和血浆中陈旧和受损的蛋白质的积累。我们假设，三种不同的增强胰岛素敏感性的模式通过不同的机制来减少陈旧和受损蛋白质的积累和DNA氧化。我们预计热量限制(CR)可以通过减少ROS的产生来减少氧化应激，从而减少对蛋白质和DNA的氧化损伤。相比之下，有氧运动增加了ROS的产生，但刺激了强大的抗氧化防御系统。此外，与CR不同的是，我们预计锻炼会增加新生蛋白质的合成，并增加旧的受损蛋白质的周转。胰岛素增敏剂可减少氧化应激，促进陈旧和受损蛋白质的清除。我们将研究腹型肥胖者，并将他们与瘦肉型参与者进行比较，以确定这些胰岛素抵抗者与瘦肉型胰岛素敏感者相比，是否积累了更多受损的蛋白质和DNA。然后，我们将确定16周的有氧运动、卡路里限制和胰岛素增敏剂与安慰剂相比，是否会减轻对蛋白质和DNA的氧化损伤，并提高胰岛素敏感性。拟议的研究将利用最先进的方法进行，其中许多方法是我们的实验室在上一个供资期间开发的。我们期望这些研究的结果除了展示体内维持功能蛋白质组的机制外，还将为胰岛素抵抗和2型糖尿病的潜在机制提供开创性的见解。 公共卫生相关性：拟议的实验调查了糖尿病和相关健康问题在社会中迅速扩大的潜在新的潜在潜在机制。第一个具体目标是确定当从低卡路里含量的饮食过渡到高能量含量的饮食时，更高效的线粒体--产生化学能的细胞的动力源--是否会对DNA和蛋白质造成不利影响。第二和第三个目标是使用一种新的方法在肥胖者和胰岛素抵抗者中测量肌肉和血浆中受损和陈旧蛋白质的积累，并确定有氧运动、热量限制和提高胰岛素敏感性和降低高胰岛素水平的药物如何改变这一潜在的有害过程。