VEGF gene expression in CNS microvascular pericyte

CNS微血管周细胞中VEGF基因的表达

• 批准号: 6719501
• 负责人: PAULA DORE-DUFFY
• 金额: $ 33.5万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6719501
• 关键词: DNA binding protein; cerebral ischemia /hypoxia; enzyme linked immunosorbent assay; flow cytometry; gene expression; high performance liquid chromatography; immunocytochemistry; laboratory rat; macrophage; neurogenetics; neuroregulation; prostaglandin endoperoxide synthase; prostaglandins; translation factor; vascular endothelial growth factors; western blottings

DESCRIPTION (provided by applicant): In response to fluctuations in environmental oxygen in the cells of the blood brain barrier (BBB) undergo a number of complex adaptive measures in order to maintain tissue homeostasis and hemostasis. These adaptive responses are particularly important when the balance of oxygen availability and utilization is altered as a result of the pathophysiology of CNS disease. At the microvascular level oxygen responsive signaling mechanisms involving both the endothelial cell and the pericyte regulate angiogenesis, vascular permeability and metabolism. We propose to investigate early responses of the CNS pericyte following in vitro exposure to hypoxia using the GaSPak 100 hypoxia system (Becton Dickinson and Company Sparks, MD.) Within 15 minutes of exposure to low oxygen (1%) pericytes synthesize and release cyclopentenone prostaglandins. Increased PGD2 and the dehydration product delta12pGJ2 were detected by HPLC and by immune techniques. Delta12pGD2 was not synthesized. Pericytes constitutively express three of the five alternate splice variants of vascular endothelial cell growth factor (VEGF) mRNA, but little to no protein. Exposure to low oxygen increased mRNA levels as well as the synthesis and release of VEGF protein. Addition of either Delta12pGJ2 or 15-deoxyDelta12/14PGJ2 to pericyte under normoxic conditions increased the synthesis and release of VEGF protein in a dose dependent manner. Results suggest that PGD produced by the CNS pericyte is an early signaling molecule in regulation of the angiogenic response to hypoxia. We will further investigate the pericyte VEGF response: Aim #l: Determine the mechanism of hypoxia induced upregulation of VEGF gene expression. We will also evaluate the role of translation initially and inhibiting factor elF-4E and 4EBprincipal investigator. Aim#2: To determine the role of COX-1 in pericyte VEGF response to hypoxia. Aim#3: To determine the role of the COX-1 mediated release of pericyte PGD/PGJ in hypoxia. We propose that the CNS microvascular pericyte is a regulatory cell important in the vascular response to hypoxic stress and that cyclopentenone prostagladins are involved in the pericyte stress response.

描述（由申请人提供）：响应血液脑屏障（BBB）细胞中环境氧气的波动，采取了许多复杂的自适应措施，以维持组织稳态和止血。当CNS疾病的病理生理学改变氧气和利用率的平衡时，这些适应性反应尤其重要。在微血管水平，氧气响应信号传导机制涉及内皮细胞和周细胞调节血管生成，血管渗透性和代谢。我们建议在暴露于低氧（1％）周围的氧气（1％）合成和释放环戊烯酮的前列腺素后15分钟内，使用Gaspak 100缺氧系统（Becton Dickinson and Company Sparks）研究中枢神经系统周细胞的早期反应。通过HPLC和免疫技术检测PGD2和脱水产物Delta12pgj2。 delta12pgd2未合成。周细胞组成性地表达血管内皮细胞生长因子（VEGF）mRNA的五个替代剪接变体中的三个，但几乎没有蛋白质。暴露于低氧增加的mRNA水平以及VEGF蛋白的合成和释放。在正氧条件下向周细胞中添加delta12pgj2或15-脱氧二硫代氏菌12/14pgj2，以剂量依赖性的方式增加了VEGF蛋白的合成和释放。结果表明，中枢神经系统周围产生的PGD是调节血管生成反应缺氧的早期信号分子。我们将进一步研究周围VEGF的响应：AIM #L：确定缺氧诱导的VEGF基因表达上调的机理。我们还将评估最初翻译和抑制因子Elf-4e和4ebprincipal研究者的作用。目标＃2：确定COX-1在周围VEGF对缺氧反应中的作用。目标＃3：确定COX-1介导的周细胞PGD/PGJ在缺氧中的作用。我们建议中枢神经系统微血管周细胞是对低氧应激反应重要的调节细胞，并且环戊烯酮前列腺素参与周细胞应力反应。