PKC Activation and Cardiovascular Disease in Diabetes

糖尿病中的 PKC 激活与心血管疾病

• 批准号: 7107973
• 负责人: GEORGE L KING
• 金额: $ 40.52万
• 依托单位: JOSLIN DIABETES CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7107973
• 关键词: angiogenesis; apolipoprotein E; atherosclerosis; cytokine; diabetic angiopathy; disease /disorder model; enzyme activity; enzyme induction /repression; genetically modified animals; glucose metabolism; insulin dependent diabetes mellitus; kinase inhibitor; laboratory mouse; leukocyte activation /transformation; lipid metabolism; monocyte; protein isoforms; protein kinase C

DESCRIPTION (provided by applicant): Risk factors for cardiovascular disease (CVD) in type 1 diabetic patients (T1DM) include hyperglycemia, hyperlipidemia, renal dysfunction, oxidants, glycated lipids and proteins and other metabolic abnormalities. The pathologies of CVD in T1DM are similar to those of nondiabetic subjects, suggesting the risk factors accelerating CVD also present in T1DM. Activation of monocytes, endothelial cells (EC) and smooth muscle cells (SMC) are shown to be important for the development of atherosclerosis both in T1DM and nondiabetic patients. Protein kinase C (PKC) activation is important for promoting these cells into pro-atherogenic state. Many of the risk factors stated above, will activate PKC in monocytes and vascular cells. Thus, we propose that abnormal metabolites of glucose and lipids due to insulin deficiency will activate PKC, especially the beta isoforms, in monocytes, EC and SMC. This will result in the expression and secretion of proteins in favor of the formation of macrophages/foam cells, endothelial dysfunction, and SMC migration and proliferation. These alterations will further accelerate atherosclerosis in T1DM. To test this hypothesis definitively, we propose: l) to characterize the role of PKC alpha, beta, and delta isoforms in glucose-induced activation of monocytes, arterial EC and SMC. 2) to assess the role of PKC beta isoform activation in the development of atherosclerosis in apoE-KO mice by comparing the atherosclerotic lesions in :a) diabetic or nondiabetic apoE-KO mice with or without PKC beta inhibitor; b) offsprings (PKC beta 2 V-TgXapoE-KO) of mice with vascular-specific overexpression of PKC beta 2 (PKC beta 2 V-Tg) crossbred with apoE KO-mice; c) offsprings of PKC beta null mice (PKC beta-KO) crossbred with apoE-KO mice. 3) to make and characterize mice overexpressing PKC beta 2 isoform in monocytes using a monocyte CD68 promoter (PKC beta 2 M-Tg) with respect to retinal, renal and cardiac abnormalities in nondiabetic and diabetic state. We will crossbreed PKC beta 2 M-Tg mice with apoE KO mice to determine its effect on the development of atherosclerosis.

描述（由申请人提供）： 1型糖尿病患者（T1DM）中心血管疾病（CVD）的危险因素包括高血糖，高脂血症，肾功能不全，氧化剂，糖化脂质和蛋白质以及其他代谢异常。 T1DM中CVD的病理与非糖尿病受试者相似，这表明T1DM中也存在加速CVD的危险因素。单核细胞，内皮细胞（EC）和平滑肌细胞（SMC）的激活对于在T1DM和非糖尿病患者中的动脉粥样硬化的发展都很重要。蛋白激酶C（PKC）激活对于将这些细胞促进促动脉粥样硬化状态很重要。上述许多危险因素将激活单核细胞和血管细胞中的PKC。因此，我们提出，由于胰岛素缺乏症引起的葡萄糖和脂质异常代谢物将激活PKC，尤其是单核细胞，EC和SMC中的β同工型。这将导致蛋白质的表达和分泌，而有利于形成巨噬细胞/泡沫细胞，内皮功能障碍以及SMC迁移和增殖。这些改变将进一步加速T1DM的动脉粥样硬化。为了确切地检验该假设，我们提出：l）表征PKCα，β和三角洲同工型在葡萄糖诱导的单核细胞激活，动脉EC和SMC中的作用。 2）通过比较以下方法，评估PKCβ同工型激活在APOE-KO小鼠中动脉粥样硬化中的作用：a）具有或没有PKCβ抑制剂的糖尿病或非糖尿病患者或非糖尿病患者APOE-KO小鼠； b）具有PKCβ2（PKC Beta 2 V-TG）的血管特异性过表达的小鼠的后代（PKC Beta 2 V-TGXAPOE-KO），用APOE KO-MICE杂交； c）PKC Beta Null小鼠（PKC Beta-KO）的后代与Apoe-KO小鼠杂交。 3）使用单核细胞CD68启动子（PKC Beta 2 M-TG）在单核细胞中过表达PKCβ2同工型的小鼠相对于视网膜，肾脏和心脏异常的非糖尿病和糖尿病状态。我们将与APOE KO小鼠交叉杂交的PKC Beta 2 M-TG小鼠，以确定其对动脉粥样硬化发展的影响。