Protective Effects of Stearic Acid on Gestational and Acquired Diabetes Mellitus

硬脂酸对妊娠期和获得性糖尿病的保护作用

• 批准号: 7289736
• 负责人: Gregory A Graf
• 金额: $ 29.16万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2010-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7289736
• 关键词: AMP-activated protein kinase kinase; Ablation; Acetyl-CoA Carboxylase; Acyl Coenzyme A; Adipocytes; Adipose tissue; Affect; Angiotensin II; Biochemical; Biochemistry; Blood Pressure; Blood Vessels; CD36 gene; Cardiovascular system; Carnitine; Cartoons; Caveolae; Cells; Coenzyme A; Commit; Data; Development; Diabetes Mellitus; Esterification; Esterified Fatty Acids; Esters; Fatty Acids; Gene Targeting; Genes; Genetic; Gestational Diabetes; Glucose; Goals; In Vitro; Individual; Insulin; Insulin Resistance; Intervention; Knock-out; Knockout Mice; Linoleic Acids; Lipids; Location; Measurement; Measures; Mediating; Membrane Microdomains; Metabolic; Mitochondria; Modeling; Mothers; Mus; Nonesterified Fatty Acids; Numbers; OGTT; Outcome; Phenotype; Phosphorylation; Pregnancy; Production; Proteins; Rate; Research Personnel; Role; Route; Signal Transduction; Stearic Acids; Testing; Thick; Tissues; Transferase; Triad Acrylic Resin; Triglycerides; Upper arm; Weight; Withdrawal; clinically relevant; concept; day; db/db mouse; diabetes mellitus genetics; diabetic; diacylglycerol O-acyltransferase; fatty acid biosynthesis; fatty acid oxidation; feeding; glucose metabolism; improved; in vivo; in vivo Model; knockout gene; novel; oral glucose tolerance; oxidation; peroxisome; programs; protective effect; uptake; AMP-activated protein kinase kinase; Ablation; Acetyl-CoA Carboxylase; Acyl Coenzyme A; Adipocytes; Adipose tissue; Affect; Angiotensin II; Biochemical; Biochemistry; Blood Pressure; Blood Vessels; CD36 gene; Cardiovascular system; Carnitine; Cartoons; Caveolae; Cells; Coenzyme A; Commit; Data; Development; Diabetes Mellitus; Esterification; Esterified Fatty Acids; Esters; Fatty Acids; Gene Targeting; Genes; Genetic; Gestational Diabetes; Glucose; Goals; In Vitro; Individual; Insulin; Insulin Resistance; Intervention; Knock-out; Knockout Mice; Linoleic Acids; Lipids; Location; Measurement; Measures; Mediating; Membrane Microdomains; Metabolic; Mitochondria; Modeling; Mothers; Mus; Nonesterified Fatty Acids; Numbers; OGTT; Outcome; Phenotype; Phosphorylation; Pregnancy; Production; Proteins; Rate; Research Personnel; Role; Route; Signal Transduction; Stearic Acids; Testing; Thick; Tissues; Transferase; Triad Acrylic Resin; Triglycerides; Upper arm; Weight; Withdrawal; clinically relevant; concept; day; db/db mouse; diabetes mellitus genetics; diabetic; diacylglycerol O-acyltransferase; fatty acid biosynthesis; fatty acid oxidation; feeding; glucose metabolism; improved; in vivo; in vivo Model; knockout gene; novel; oral glucose tolerance; oxidation; peroxisome; programs; protective effect; uptake

DESCRIPTION (provided by applicant): Hypothesis. The central hypothesis of this proposal is that CD36, ABCD2, and mitoNEET mediate the protective effects of stearic acid on gestational diabetes and on the acquired diabetes of the progeny. Rationale. The goal of this proposal is to test the novel hypothesis that stearic acid ameliorates gestational diabetes mellitus (GDM) and subsequent diabetes in the progeny of GDM mothers through a triad of CD36, ABCD2, and mitoNEET proteins. We will use the CSTBLKS/J-Lep/*'* mouse that develops GDM around day 16 of gestation. We will examine the mechanisms of stearic acid on CD36, ABCD2, and mitoNEET expression and function in both GDM and on long-term metabolic and cardiovascular outcomes in the progeny of the db/+ dams. Finally, we will use individual genetic knockout models for each protein to determine the extent to which each protein mediates the action of stearic acid in both GDM and diabetes of the progeny. The uniqueness and potential clinical relevance of this approach is the mechanistic analysis of a possible means for treatment of both GDM and pursuant diabetes mellitus acquired by the offspring. We propose in vivo studies that can be further dissected by in vitro studies with isolated tissues and cells, which will enable us to thoroughly explore the protective mechanism of action of stearic acid. 1. To determine the impact of stearic acid on the development and extent of GDM. Db/+ dams will be used to examine whether stearic acid affects the development of GDM by measuring oral glucose tolerance, vascular responsiveness, and adipocyte function. The expression and subcellular localization of CD36, ABCD2, and mitoNEET in adipose will be quantified and functionality of the proteins assessed through measurements of fatty acid biochemistry (p-oxidation, fatty acid composition, and angiotensin II secretion). Similarly, we will also examine the effect of stearic acid treatment fed exclusively to the db/+ dams on the extent of diabetes in the offspring. Finally, we will thoroughly examine the mechanism of action of stearic acid on fatty acid biochemistry in insulin resistant 3T3-L1 adipocytes. This aim is important because we will mechanistically evaluate a potential palliative intervention for both GDM and the accompanying diabetes of the offspring. 2. To determine the duration which stearic acid improves the diabetic phenotype in db/db mice. Db/db progeny will be treated with stearic acid and the duration and extent of the protective effect investigated. The effect of withdrawal of stearic acid on the lasting benefit of the intervention will also be examined. We will measure blood pressure, vascular responsiveness, weight and glucose. We will examine the mechanisms of stearic acid action by determining the expression and subcellular location of CD36, ABCD2, and mitoNEET in adipose and assessing function of each protein through measurements of adipose fatty acid biochemistry. This aim is important because it will provide data on the cardiovascular benefit and sustainability of stearic acid intervention. 3. To determine whether the beneficial effects of stearic acid are mediated by CD36, ABCD2, and/or mitoNEET. We will cross mice null for CD36, ABCD2, or mitoNEET with db/+ mice and assess the efficacious effects of stearic acid in ameliorating GDM and the ensuing diabetic phenotype of the double crossed db/db progeny lacking each individual gene. We will measure OGTT, vascular responsiveness, and adipose fatty acid biochemistry in db/+ dams. Furthermore, we will assess the effect of individual gene knockout on the ability of stearic acid to affect the development of diabetes in the db/db mice. This will be accomplished through measurements of weight, glucose, insulin, blood pressure, and vascular responsiveness. Mechanistically, the effects of gene ablation will be dissected through comprehensive analyses of isolated primary adipose biochemistry. This aim is critical because each double crossed mouse line will provide the definitive mechanistic data regarding the role of each protein in mediating the beneficial effects of stearic acid.

描述（由申请人提供）：假设。该提案的中心假设是CD36，ABCD2和Mitoneet介导了硬脂酸对妊娠糖尿病和后代获得的糖尿病的保护作用。理由。该提案的目的是检验新的假设，即硬脂酸可以通过CD36，ABCD2和Mitoneet蛋白质的三合会来改善GDM母亲后代的妊娠糖尿病（GDM）和随后的糖尿病。我们将使用CSTBLKS/J-LEP/*'*鼠标，该鼠标在妊娠第16天左右开发GDM。我们将在GDM和DB/+大坝后代的GDM以及长期代谢和心血管结局中检查CD36，ABCD2和Mitoneet表达和功能的硬脂酸的机制。最后，我们将为每种蛋白质使用单个遗传基因敲除模型来确定每种蛋白在后代的GDM和糖尿病中介导硬脂酸的作用的程度。这种方法的独特性和潜在临床相关性是对后代获得的GDM和根据糖尿病的可能手段的机械分析。我们提出了可以通过分离的组织和细胞进行体外研究进一步阐述的体内研究，这将使我们能够彻底探索硬脂酸作用的保护机理。 1。确定硬脂酸对GDM的发育和程度的影响。 DB/+大坝将用于检查硬脂酸是否通过测量口服葡萄糖耐受性，血管反应性和脂肪细胞功能来影响GDM的发展。脂肪中CD36，ABCD2和Mitoneet的表达和亚细胞定位将被量化，并通过测量脂肪酸生物化学（P-氧化，脂肪酸组成和血管素II分泌）评估蛋白质的功能。同样，我们还将检查仅喂入DB/+大坝对后代糖尿病程度的硬脂酸处理的影响。最后，我们将彻底检查硬脂酸对胰岛素耐药3T3-L1脂肪细胞中脂肪酸生物化学的作用机理。这个目标很重要，因为我们将对后代的GDM和随附的糖尿病进行机械学评估潜在的姑息干预措施。 2。确定硬脂酸改善db/db小鼠中糖尿病表型的持续时间。 DB/DB后代将用硬脂酸和研究的保护作用的持续时间和程度处理。还将检查戒断硬脂酸对干预效果的持久益处的影响。我们将测量血压，血管反应能力，体重和葡萄糖。我们将通过确定脂肪中CD36，ABCD2和mitoneet的表达和亚细胞位置来检查硬脂酸作用的机制，并通过测量脂肪脂肪酸生物化学的测量来评估每种蛋白质的功能。该目标很重要，因为它将提供有关硬脂酸干预的心血管益处和可持续性的数据。 3。确定硬脂酸的有益作用是否是由CD36，ABCD2和/或Mitoneet介导的。我们将与DB/+小鼠的CD36，ABCD2或Mitoneet无效，并评估硬脂酸在改善GDM的有效作用以及随之而来的双交叉DB/DB后代的糖尿病表型中缺乏每个基因的糖尿病表型。我们将在DB/+大坝中测量OGTT，血管反应性和脂肪脂肪酸生物化学。此外，我们将评估单个基因基因敲除对硬脂酸影响DB/DB小鼠糖尿病发展的能力的影响。这将通过测量体重，葡萄糖，胰岛素，血压和血管反应能力来实现。从机械上讲，将通过对孤立的原代脂肪生物化学的全面分析进行基因消融的影响。该目标至关重要，因为每条双重小鼠系将提供有关每种蛋白质在介导硬脂酸的有益作用中的作用的确定机械数据。