12/15-Lipoxygenase and Diabetic Neuropathy

12/15-脂氧合酶和糖尿病神经病变

基本信息

批准号：
7314920
负责人：
IRINA G OBROSOVA
金额：
$ 31.45万
依托单位：
LSU PENNINGTON BIOMEDICAL RESEARCH CTR
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：
2007-09-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7314920
关键词：
12-HETE 12-HPETE Acids Adult Afferent Neurons Aldehyde Reductase Animal Model Animals Arachidonate 15-Lipoxygenase Arachidonic Acids Atrophic Behavioral Biochemical Biological Assay Cells Cholesterol Esters Coculture Techniques Complications of Diabetes Mellitus Condition Cultured Cells Cytochrome P450 Data Development Diabetes Mellitus Diabetic Neuropathies Diabetic mouse Diet Dioxygenases Enzymes Epithelial Cells Exposure to Fatty acid glycerol esters Free Radicals Functional disorder Gene Expression Glucose High Pressure Liquid Chromatography Human Inflammatory Response Iron Kidney Kidney Diseases Leukotrienes Link Lipid Peroxidation Lipids Lipoproteins Lipoxins Lipoxygenase Lipoxygenase 1 Lipoxygenase Inhibitors Metabolic Pathway Metabolism Mitogen-Activated Protein Kinases Modeling Molecular Mus Neurons Neuropathy Nuclear Pathogenesis Peripheral Peripheral Nerves Phospholipids Phosphorylation Physiological Polymerase Chain Reaction Prevention Production Prostaglandins Protein Overexpression Retinal Diseases Role Scheme Schwann Cells Severities Signal Transduction Site Small Interfering RNA Smooth Muscle Spinal Cord Streptozocin Stress Time Tissues Transcription Factor AP-1 Transfection Tubular formation Up-Regulation Western Blotting Wild Type Mouse cyclooxygenase 1 diabetic exposed human population feeding in vivo indexing interest macrophage member monocyte mouse model podocyte type I and type II diabetes

项目摘要

DESCRIPTION (provided by applicant): Evidence for the important role of arachidonic acid metabolism in diabetes complications is emerging. Recent in vivo and cell culture studies including those from our group revealed that 12/15-lipoxygenase (12/15-LO) 1) contributes to impaired cell signaling and inflammatory response; and 2) is implicated in the pathogenesis of endothelial dysfunction, nephropathy and retinopathy, associated with diabetes. The objective of this proposal is to evaluate the role of 12/15-LO in peripheral diabetic neuropathy (PDN) using animal models of Type 1 and Type 2 diabetes and high glucose-exposed co-cultures of mouse Schwann cells and DRG neurons and cultures of human Schwann cells (HSC). Our preliminary data indicate that 1) 12/15-LO is abundantly expressed in the peripheral nerve and its expression and activity increase in diabetic conditions; 2) 12/15-LO-/- mice develop less severe PDN than wild-type mice; 3) some manifestations of PDN in STZ-diabetic mice are reversed by a short-term 12/15-LO inhibitor treatment; 4) 12/15-LO overexpression is manifest after a short-term (24h) exposure of HSC to high glucose; and 5) a 12/15-LO inhibitor treatment counteracts high glucose-induced mitogen-activated protein kinase (MAPK) phosphorylation in HSC. The SPECIFIC AIMS are 1) evaluate two structurally unrelated 12/15-LO inhibitors on functional, biochemical and structural indices of PDN in mouse models of Type 1 and Type 2 diabetes, i.e., STZ-diabetic and high-fat diet fed mice; 2) compare severity of PDN in 12/15-LO-/- mice and wild-type mice with Type 1 and Type 2 diabetes; 3) assess the role for the most important, "upstream", mechanism of PDN, i.e. increased aldose reductase activity, in 12/15-LO upregulation, and 4) examine the contribution of 12/15-LO to MAPK activation, in peripheral nerve, spinal cord and DRG neurons in the afore-mentioned animal and cell culture models. The project will combine physiological, behavioral, biochemical, immunohistochemical and structural studies in animals with biochemical (HPLC, Western blotting, spectrofluorometric enzymatic assays) and molecular (real-time PCR, siRNA transfections) approaches in cell culture models. The findings will generate new information on the role for 12/15-LO in PDN of Type 1 and Type 2 diabetes, and may provide rationale for development of 12/15-LO inhibitors for its prevention and treatment.

描述（由申请人提供）：花生四烯酸代谢在糖尿病并发症中的重要作用的证据正在出现。最近的体内和细胞培养研究（包括我们小组的研究）表明，12/15-脂氧合酶 (12/15-LO) 1) 会导致细胞信号传导和炎症反应受损； 2) 参与与糖尿病相关的内皮功能障碍、肾病和视网膜病的发病机制。该提案的目的是使用 1 型和 2 型糖尿病动物模型以及小鼠雪旺细胞和 DRG 神经元和培养物的高葡萄糖暴露共培养物来评估 12/15-LO 在周围糖尿病神经病变 (PDN) 中的作用人类雪旺细胞 (HSC)。我们的初步数据表明：1）12/15-LO在周围神经中大量表达，并且在糖尿病状态下其表达和活性增加； 2) 12/15-LO-/- 小鼠的 PDN 比野生型小鼠严重程度较轻； 3)短期12/15-LO抑制剂治疗可以逆转STZ糖尿病小鼠中PDN的一些表现； 4) HSC短期（24小时）暴露于高糖后明显出现12/15-LO过表达； 5) 12/15-LO抑制剂治疗可抵消HSC中高葡萄糖诱导的丝裂原激活蛋白激酶(MAPK)磷酸化。具体目标是 1) 评估两种结构上不相关的 12/15-LO 抑制剂对 1 型和 2 型糖尿病小鼠模型（即 STZ 糖尿病小鼠和高脂饮食喂养小鼠）中 PDN 的功能、生化和结构指标的影响； 2) 比较患有1型和2型糖尿病的12/15-LO-/-小鼠和野生型小鼠的PDN严重程度； 3) 评估 PDN 最重要的“上游”机制，即醛糖还原酶活性增加，在 12/15-LO 上调中的作用，以及 4) 检查 12/15-LO 对外周血中 MAPK 激活的贡献上述动物和细胞培养模型中的神经、脊髓和DRG神经元。该项目将把动物的生理、行为、生化、免疫组织化学和结构研究与细胞培养模型中的生化（HPLC、蛋白质印迹、荧光分光酶测定）和分子（实时 PCR、siRNA 转染）方法结合起来。这些发现将产生关于 12/15-LO 在 1 型和 2 型糖尿病 PDN 中的作用的新信息，并可能为开发 12/15-LO 抑制剂用于预防和治疗提供理论依据。