PKC Activation and Cardiovascular Disease in Diabetes

描述（由申请人提供）： 1型糖尿病（T1 DM）患者心血管疾病（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。T1 DM中CVD的病理学与非糖尿病受试者相似，表明加速CVD的风险因素也存在于T1 DM中。单核细胞、内皮细胞（EC）和平滑肌细胞（SMC）的活化在T1 DM和非糖尿病患者动脉粥样硬化的发展中均显示出重要性。蛋白激酶C（PKC）的激活对于促进这些细胞进入促动脉粥样硬化状态是重要的。上述许多危险因素都会激活单核细胞和血管细胞中的PKC。因此，我们提出，由于胰岛素缺乏导致的葡萄糖和脂质代谢异常将激活单核细胞、EC和SMC中的PKC，尤其是β亚型。这将导致有利于巨噬细胞/泡沫细胞形成、内皮功能障碍以及SMC迁移和增殖的蛋白质的表达和分泌。这些改变将进一步加速T1 DM患者的动脉粥样硬化。为了验证这一假设，我们提出：1）PKC α，β和δ亚型在葡萄糖诱导的单核细胞，动脉EC和SMC活化中的作用。2)通过比较以下小鼠的动脉粥样硬化病变来评估PKC β亚型活化在apoE-KO小鼠中动脉粥样硬化发展中的作用：a）有或没有PKC β抑制剂的糖尿病或非糖尿病apoE-KO小鼠; B）后代（PKC β 2 V-TgXapoE-KO）的小鼠的血管特异性PKC β 2过表达c）与apoE KO小鼠杂交的PKC β缺失小鼠（PKC β-KO）的后代。3)制备和表征使用单核细胞CD 68启动子在单核细胞中过表达PKC β 2同种型（PKC β 2 M-Tg）的小鼠，其与非糖尿病和糖尿病状态下的视网膜、肾脏和心脏异常有关。我们将PKC β 2 M-Tg小鼠与apoE基因敲除小鼠杂交，以确定其对动脉粥样硬化发展的影响。

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.