EICOSANOIDS, VASCULAR TONE AND BLOOD PRESSURE

类花生酸、血管张力和血压

• 批准号: 6627521
• 负责人: WILLIAM BRYSON CAMPBELL
• 金额: $ 29.9万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-01 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6627521
• 关键词: arachidonate; chemical structure function; eicosanoid metabolism; eicosanoids; gas chromatography mass spectrometry; high performance liquid chromatography; hormone regulation /control mechanism; immunologic assay /test; laboratory rabbit; lipoxygenase; muscle tone; nitric oxide; potassium channel; stereoisomer; tissue /cell culture; vascular endothelium; vascular smooth muscle; vasoactive agent; vasodilation; vasodilators; vasomotion

DESCRIPTION (Verbatim from the application): The importance of the endothelial cell (EC) lining of the blood vessel in the regulation of vascular tone is clearly established. Hormones such as acetylcholine, substance P and bradykinin relax blood vessels in the presence of the endothelium but not in its absence. This relaxation is attributed to the release of endothelium-derived relaxing factor (EDRF). There is now evidence for multiple EDRFs. One is nitric oxide (termed NO-EDRF). In addition, there is an EDRF released by arachidonic acid (AA-EDRF) that is not PGI2 but a lipoxygenase metabolite. We have identified this AA-EDRF as a metabolite of 1 5-lipoxygenase, 11(S),12(S), 1 5(S)-trihydroxyeicosatrienoic acid (11,12,1 5-THETA), using bioassay, HPLC, chemical synthesis and gas chromatography/mass spectrometry. 15(S)-Hydroxy-1 1,12-epoxyeicosatrienoic acid (15-H-i i,12-EETA) is thought to be the precursor of 11,12,15-THETA. Both 15-H-i 1,12-EETA and 1 1(S),12(S),15(S)-THETA relax the rabbit aorta and one or both represent AA-EDRF. The proposed studies will test the hypothesis that arachidonic acid is metabolized by the endothelium to vasodilator eicosanoids, 15-H-i 1,12-EETA and 11,12,15-THETA, that are involved in the regulation of vascular tone and contribute to the action of vasoactive hormones. Studies will be conducted in isolated blood vessels, smooth muscle cells and ECs. Using chemical, analytical, biochemical, electrophysiological and pharmacological approaches, the proposed experiments will 1. determine if 15-H-11,12-EETA or other HEETAs are produced by the aorta and ECs and determine the biologically active stereoisomer of 15(S)-H-11,12-EETA. Experiments will compare the vasodilator activity of the biological and synthetic stereoisomers. 2. develop an assay for 11,12,15-THETA and investigate the regulation of 15-H-11,12-EETA and 11,12,15-THETA release by vasoactive agents and inhibitors of arachidonic acid metabolism. This quantitative data will be combined with physiological and pharmacological studies to determine the contribution of the THETA and HEETA to vascular tone and the activity of vasoactive hormones. 3. study the role of 15-lipoxygenase-1 and -2 in the biosynthesis of 15-H-11,12-EETA and 11,12,15-THETA and their degradation by 15-hydroxy PG dehydrogenase using purified enzymes, antibodies and inhibitors. 4. investigate the mechanism of smooth muscle dilation by 15-H-11,12-EETA and 11,12,15-THETA by determining their action on membrane potential and potassium channel activity.

描述(来自应用程序的逐字)：内皮的重要性 细胞(EC)衬里对血管张力的调节作用 很明显已经确定了。乙酰胆碱、P物质和缓激肽等激素 在有内皮的情况下放松血管，但不要在没有内皮的情况下放松血管。 这种松弛归因于内皮衍生的松弛的释放。 系数(EDRF)。现在有证据表明存在多个EDRF。一种是一氧化氮 (称为非EDRF)。此外，还有花生四烯酸释放的EDRF (AA-EDRF)，不是PGI2，而是脂氧合酶代谢产物。我们已经确定了 此AA-EDRF为15-脂氧合酶的代谢产物，11(S)，12(S)，1 5(S)-三羟基二十碳三烯酸(11，12，15-theta)，采用生物测定，高效液相色谱法， 化学合成和气相色谱/质谱联用。15(S)-羟基-1 1，12-环氧二十碳三烯酸(15-H-I，12-EETA)被认为是其前体 11，12，15-Theta。15-H-I 1，12-EETA和11(S)，12(S)，15(S)-西塔放松 兔主动脉和一个或两个代表AA-EDRF。拟议中的研究将检验 花生四烯酸由血管内皮细胞代谢到 血管扩张剂二十烷类化合物，15-H-I，1，12-EETA和11，12，15-Theta 在调节血管张力和参与血管活性活动中起作用 荷尔蒙。研究将在分离的血管、平滑肌肉中进行 细胞和内皮细胞。使用化学、分析、生化、电生理学 和药理学方法，拟议的实验将1.确定是否 15-H-11，12-EETA或其他HEETA由主动脉和内皮细胞产生，并确定 15(S)-H-11，12-EETA的生物活性立体异构体。实验将会 比较生物和合成立体异构体的血管扩张活性。 2.建立了11，12，15-theta的测定方法，并研究了 血管活性物质和抑制剂释放15-H-11，12-EETA和11，12，15-Theta 花生四烯酸新陈代谢。这一量化数据将与 生理学和药理学研究，以确定 Theta和HEETA对血管张力和血管活性激素活性的影响。3. 15-脂氧合酶-1和-2在生物合成中的作用研究 15-H-11，12-EETA和11，12，15-Theta及其在15-羟基PG中的降解 使用纯化的酶、抗体和抑制剂进行脱氢酶。4.调查 15-H-11，12-EETA和11，12，15-Theta的血管扩张机制 通过测定它们对膜电位和钾通道的作用 活动。