P450 EICOSANOIDS, CELL CALCIUM AND VASCULAR FUNCTION

P450 类二十烷酸、细胞钙和血管功能

• 批准号: 6125832
• 负责人: EARL F ELLIS
• 金额: $ 30.01万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6125832
• 关键词: biological signal transduction; brain circulation; calcium flux; calcium indicator; cytochrome P450; eicosanoid metabolism; eicosanoids; gas chromatography mass spectrometry; high performance liquid chromatography; intracellular transport; laboratory rat; paracrine; phospholipase A2; radiotracer; thapsigargin; tissue /cell culture; ultrasound blood flow measurement; vascular smooth muscle; vasodilation; vasomotion; voltage /patch clamp

DESCRIPTION: (Adapted from the application) Recent evidence has suggested that Calcium Influx Factor (CIF), an unidentified factor which signals influx of extracellular calcium in response to depletion of intracellular calcium stores, is 5,6-epoxyeicosatrienoic acid (5,6-EET), a P450 monooxygenase metabolite of arachidonic acid (AA). The proposed studies will further test the hypothesis that P450 metabolites of AA formed upon depletion of Ca2+ stores induce influx of extracellular Ca2+, and thus influence cell function and intercellular signaling. The hypothesized relationships will be examined in cultured astrocytes, endothelial cells and vascular smooth muscle (VSM) cells as well as in anesthetized rats. They will determine whether agonist-or thapsigargin-induced store depletion activates phospholipase A2, liberates AA and induces 5,6-EET formation with subsequent influx of Ca2+. They will also elucidate whether 5,6-EET is released from stimulated astrocytes and endothelium and acts as a paracrine signal for VSM causing activation of Ca2+-dependent K+ channels, hyperpolarization and relaxation. The hypothesized relationships will be tested in cultured cells using microspectrofluorometry with the calcium indicator Fura-2, radiolabeled tracers, HPLC, gas chromatography/mass spectrometry and patch clamping. Tests of applicability to the control of the cerebral circulation will employ the in vivo cranial window technique, microscopy and laser-Doppler flowmetry. These studies will provide important basic information relevant to the role of P450 eicosanoids in the regulation of intra- and intercellular relationships of many cell types, including those which control brain blood flow.

描述：（从应用程序改编）最新证据提出了 钙涌入因子（CIF），这是一个信号的未识别因子 响应细胞内耗竭的细胞外钙的流入 钙存储，为5,6-蛋白酶钠酸（5,6-EET），P450 蛛网膜酸（AA）的单加氧酶代谢产物。 提出的研究 将进一步检验以下假设： Ca2+存储的耗竭诱导细胞外Ca2+的涌入，因此 影响细胞功能和细胞间信号。 假设 关系将在培养的星形胶质细胞，内皮细胞和 血管平滑肌（VSM）细胞以及麻醉大鼠。 他们 将确定激动剂或thapsigargin诱导的商店耗尽 激活磷脂酶A2，释放AA并诱导5,6-EET形成 随后的Ca2+流入。 他们还将阐明是否5,6-EET是 从刺激的星形胶质细胞和内皮中释放，并用作旁分泌 VSM的信号导致Ca2+依赖性K+通道的激活， 超极化和放松。 假设的关系将是 使用钙测试在培养的细胞中测试 指标Fura-2，放射性标记的示踪剂，HPLC，气相色谱/质量 光谱和贴片夹具。 对控制的适用性测试 脑循环将采用体内颅窗技术， 显微镜和激光多普勒流量指标。 这些研究将提供 与p450类eicosanoids在 调节许多细胞类型的细胞内和细胞间关系， 包括控制脑血流的那些。