Protective Effects of Stearic Acid on Gestational and Acquired Diabetes Mellitus

描述（由申请人提供）：假设。该建议的中心假设是，CD 36，ABCD 2和mitoNEET介导硬脂酸对妊娠糖尿病和后代获得性糖尿病的保护作用。理由。该提案的目的是检验新的假设，即硬脂酸通过CD 36、ABCD 2和mitoNEET蛋白三联体改善妊娠期糖尿病（GDM）和GDM母亲后代的后续糖尿病。我们将使用在妊娠第16天左右发生GDM的CSTBLKS/J-Lep/L1小鼠。我们将研究硬脂酸对GDM中CD 36、ABCD 2和mitoNEET表达和功能的作用机制，以及对db/+母鼠后代长期代谢和心血管结局的作用机制。最后，我们将使用每种蛋白质的个体遗传敲除模型来确定每种蛋白质在多大程度上介导硬脂酸在后代GDM和糖尿病中的作用。这种方法的独特性和潜在的临床相关性是对治疗GDM和后代获得的继发性糖尿病的可能方法的机制分析。我们建议在体内研究，可以进一步解剖在离体组织和细胞的体外研究，这将使我们能够彻底探索硬脂酸的保护作用机制。1.确定硬脂酸对GDM发展和程度的影响。Db/+母鼠将用于通过测量口服葡萄糖耐量、血管反应性和脂肪细胞功能来检查硬脂酸是否影响GDM的发展。将定量脂肪中CD 36、ABCD 2和mitoNEET的表达和亚细胞定位，并通过测量脂肪酸生物化学（β-氧化、脂肪酸组成和血管紧张素II分泌）评估蛋白质的功能。同样，我们还将检查仅对db/+母鼠饲喂硬脂酸处理对后代糖尿病程度的影响。最后，我们将深入研究硬脂酸对胰岛素抵抗3 T3-L1脂肪细胞中脂肪酸生物化学的作用机制。这一目标是重要的，因为我们将机械地评估一个潜在的姑息干预GDM和伴随糖尿病的后代。2.确定硬脂酸改善db/db小鼠糖尿病表型的持续时间。将用硬脂酸处理Db/db后代，并研究保护作用的持续时间和程度。还将检查停止硬脂酸对干预的持久益处的影响。我们将测量血压、血管反应性、体重和血糖。我们将通过测定CD 36、ABCD 2和mitoNEET在脂肪中的表达和亚细胞位置，并通过测量脂肪脂肪酸生物化学来评估每种蛋白质的功能，来研究硬脂酸作用的机制。这一目标很重要，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由CD 36、ABCD 2和/或mitoNEET介导。我们将CD 36、ABCD 2或mitoNEET缺失的小鼠与db/+小鼠杂交，并评估硬脂酸在改善GDM和缺乏每个单独基因的双杂交db/db后代的糖尿病表型方面的有效作用。我们将测量db/+母鼠的OGTT、血管反应性和脂肪脂肪酸生化。此外，我们将评估单个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、血压和血管反应性来实现。从机制上讲，基因消融的影响将通过对分离的初级脂肪生物化学的综合分析来剖析。这一目标是至关重要的，因为每个双杂交小鼠品系将提供关于每种蛋白质在介导硬脂酸有益作用中的作用的确定性机制数据。

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.