Cell Biology of Diabetic Microvascular Complications

糖尿病微血管并发症的细胞生物学

基本信息

批准号：
7805460
负责人：
GEORGE L KING
金额：
$ 41.09万
依托单位：
JOSLIN DIABETES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7805460
关键词：
1,2-diacylglycerol Adverse effects Affect Animals Apoptosis Arteries Basement membrane Biochemical Blood Vessels Blood flow Capillary Permeability Cardiac Cardiovascular Pathology Cardiovascular system Cell Adhesion Molecules Cell model Cells Cellular biology Clinical Clinical Research Clinical Trials Data Development Diabetes Mellitus Diabetic Angiopathies Diabetic Retinopathy Diglycerides Endothelial Cells Endothelin-1 Enzyme Activation Functional disorder Glomerular Filtration Rate Glucose Hyperglycemia Kidney Kidney Glomerulus Laboratories Lesion Leukocytes MAP Kinase Gene MAPK14 gene Mediating Membrane Proteins Memory Metabolic Mitogen-Activated Protein Kinases Modeling Mus Myelin P2 Protein Myocardium NADPH Oxidase Oxidative Stress Pathogenesis Pathology Pathway interactions Patients Pericytes Phase Phenotype Physiological Platelet-Derived Growth Factor Play Production Protein Biosynthesis Protein Isoforms Protein Kinase C Proteins Proteinuria Proto-Oncogene Proteins c-akt Renal Plasma Flow Renal function Reporting Resolution Retina Retinal Retinal Diseases Role Signal Transduction Signaling Molecule Testing Tight Junctions Tissues Transforming Growth Factors Transgenic Mice Vascular Endothelial Growth Factors Vascular Permeabilities Visual Acuity angiogenesis animal data base cadherin 5 capillary bed citrate carrier cytokine diabetic diabetic patient genetic regulatory protein hemodynamics inhibitor/antagonist macular edema monocyte occludin overexpression prevent promoter retina blood vessel structure ruboxistaurin transcription factor

项目摘要

DESCRIPTION (provided by applicant): We proposed that many of the retinal, renal and cardiovascular pathologies in diabetes are due to the activation of diacylglycerol (DAG) and protein kinase C (PKC), especially ¿1/2 and d isoform pathways induced by hyperglycemia (HG). A large body of evidence in cultured vascular cells and vascular tissues (retinal, renal, glomeruli, arteries and myocardium) from diabetic animals and patients have shown that DAG/ PKC pathways are activated and associated with increases in cytokine expression or activities (VEGF, ET-1, PDGF ¿ orTGF ¿, ICAM's), capillary permeability, matrix protein production, changes in signaling molecules (MAP kinases and AKT) and elevation of oxidative stress via NADPH oxidase activation. Clinical studies have shown that PKC ¿ isoform selective inhibitor ruboxistaurin (RBX) preserved visual acuity, renal function and endothelial functions in the presence of hyperglycemia. However, for diabetic retinopathy (DR), treatment with RBX did not prevent the progression to proliferative retinopathy, but accelerated the resolution of macular edema. Both basic and clinical results suggest that PKC ¿ isoform activation may selectively affect endothelial functions. We have provided preliminary data to show that hyperglycemia enhanced pericyte apoptosis, which parallels with PKC d isoform and p38 MAPK activation, remained elevated even after the cells were returned to normal glycemic condition. Thus, these results suggest that the pericyte cell model could be a model of "Metabolic Memory" as observed in clinical studies. Thus, we are proposing that the differential activation of PKC ¿ 1/2 and d isoforms induced by hyperglycemia, are causing different and specific cellular changes in retinal endothelial cells and pericytes, respectively. Thus, to prevent the progression of DR, both PKC ¿ 1/2 and d isoforms may need to be normalized. To test this new hypothesis, we will: (1) identify the specific signaling mechanisms by which PKC ¿ 1/2 and d isoforms are mediating their different effects in retinal endothelial cells (activation of ERK1/2 MAPK-occludin and others) and pericytes (p38 MAPK, NADPH oxidases) ¿ and others; (2) define the isoform selective roles of PKC ¿ 1/2 and d isoforms on the various changes in biochemical, hemodynamic and pathological parameters observed in the endothelial cells and pericytes by using control and diabetic PKC ¿-/- and d-/- mice and endothelial cell targeted overexpression of PKCP2 in C57/BL6 mice. Finally, the idea of dual activation of PKC ¿ / d isoforms are needed to induce DR will be tested by treating diabetic PKC d-/- mice with PKC ¿ specific inhibitor RBX to determine whether most of the retinal pathologies in diabetes can be prevented.

描述（由应用提供）：我们提出，糖尿病中的许多视网膜，肾脏和心血管病理学是由于二酰基甘油（DAG）和蛋白激酶C（PKC）的激活，尤其是高血糖症（HG）诱导的1/2和Disoform途径。来自糖尿病动物和患者的培养的血管细胞和血管组织（视网膜，肾脏，肾小球，动脉和心肌）的大量证据表明，DAG/ PKC途径被激活，并且与细胞因子表达或活性的增加相关（VEGF，ET-1，PDGF。信号分子（MAP激酶和AKT）和通过NADPH氧化酶激活的氧化应激升高。临床研究表明，在存在高血糖的情况下，PKC - 同工型选择性抑制剂Ruboxistaurin（RBX）保留的视力，肾功能和内皮功能。但是，对于糖尿病性视网膜病（DR），用RBX治疗并不能阻止进展增殖，而是加速了黄斑水肿的分辨率。碱性和临床结果都表明PKC�同工型激活可能会选择性地影响内皮功能。我们提供了初步数据，以表明高血糖增强的周细胞凋亡与PKC D同工型和p38 MAPK激活相关，即使在细胞恢复正常血糖状态后，也保持升高。这是这些结果表明，如临床研究中观察到的那样，周细胞模型可能是“代谢记忆”的模型。这就是我们提出的，高血糖诱导的PKC€1/2和D同工型的差异激活分别引起视网膜内皮细胞和周细胞周围的不同和特定的细胞变化。为了防止DR的进展，可能需要将PKC€1/2和D同工型归一化。为了检验这一新的假设，我们将：（1）确定PKC€1/2和D同工型在视网膜内皮细胞（ERK1/2 MAPK-OCCLUDIN等激活）和周围（P38 MAPK，NADPH OXIDES的激活）中介导其不同的作用的特定信号传导机制。 (2) define the isoform selective roles of PKC ¿ 1/2 and d isoforms on the various changes in biochemical, hemodynamic and pathological parameters observed in the endothelial cells and pericytes by using control and diabetic PKC ¿ -/- and d-/- mice and endothelial cell targeted overexpression of PKCP2 in C57/BL6 mice.最后，将需要通过PKC»特定的抑制剂RBX处理诱导DR的PKC？ / D同工型双重激活的想法，以确定是否可以预防糖尿病中的大多数视网膜病理。