CHARACTERIZATION OF THE LUNG PGDH

肺 PGDH 的特征

• 批准号: 3320863
• 负责人: Richard T. Okita
• 金额: $ 10.3万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1992-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3320863
• 关键词: acrolein; antibody; eicosanoid metabolism; eicosanoids; electron microscopy; enzyme inhibitors; enzyme mechanism; female; gel electrophoresis; guinea pigs; human tissue; hydroxy fatty acid; immunochemistry; laboratory rabbit; lung; oxidoreductase; placenta; pregnancy; prostaglandin E; prostaglandin F; prostaglandins

The NAD+-dependent prostaglandin dehydrogenase (PGDH) catalyzes the oxidation of the 15-hydroxyl group of prostaglandins of the E and F series. This reaction represents the major route of inactivation for these reactive compounds. 15-Hydroxy compounds that did not posses the cyclopentane ring of prostaglandins also were metabolized by this enzyme. 15- Hydroxy-eicosatetraenoic acid (15-HETE) was observed to be a substrate of the lung PGDH. The PGDH has been purified from the lung of pregnant rabbits and will be used to study the metabolism of 15-HETE and other 15-hydroxy-di- and triHETEs. 15-HETE may have important biological roles in modulating 5-and 12- lipoxygenase activities of white cells and mucus secretion in the lung, thus the metabolism of 15-HETE may be important in regulating its activity. The metabolism of other omega 6hydroxy- fatty acids that contain 17- and 18-carbon atoms also will be studied. Isolated lung perfusion studies will be performed to determine if the metabolism of 15-HETE may occur in the whole lung. The enzymatic activity of the lung PGDH is induced 20- to 40-fold during pregnancy of the rabbits but our studies indicate that the amount of immunoreactive protein does not increase. An inhibitory factor of the lung PGDH was isolated from the lungs of pregnant rabbits. The inhibitor migrated with the neutral lipid fraction and the evidence suggests it consists of free fatty acids. This factor will be examined further to determine its identity and to determine if the increase in PGDH activity during pregnancy is due to changes in this inhibitor. The presence of this inhibitor is important in understanding the expression and regulation of the PGDH activity. An antibody to the PGDH has been made and will be used to localize the enzyme by immunocytochemical procedures at the light and electron microscope levels in the lung and other rabbit tissues. The anti-lung PGDH also will be used to characterize and compare the immunochemical properties of PGDH enzymes from various tissues. PGDH activity is inhibited by substances in cigarette smoke and this loss of enzymatic activity may contribute to the vascular and airway changes observed in cigarette smokers. The effects of acrolein, a reactive aldehyde in cigarette smoke, will be studied on PGDH activity. The studies described in this proposal will lead to a greater characterization of this enzyme and the function of prostaglandins and other eicosanoid derivatives in pulmonary function, in pregnancy, and in pulmonary and cardiovascular diseases.

NAD+ 依赖性前列腺素脱氢酶 (PGDH) 催化前列腺素 15-羟基的氧化 E 和 F 系列。 该反应代表了主要路线 这些活性化合物的失活。 15-羟基 不具有环戊烷环的化合物 前列腺素也通过这种酶代谢。 15- 据观察，羟基二十碳四烯酸 (15-HETE) 肺PGDH的底物。 PGD​​H 已纯化自 怀孕兔子的肺，将用于研究 15-HETE 和其他 15-羟基-二-和三HETE 的代谢。 15-HETE 可能在调节 5-和 12-白细胞的脂氧合酶活性和粘液分泌 肺，因此 15-HETE 的代谢可能很重要 调节其活动。 其他 omega 6 羟基的代谢 含有 17 和 18 个碳原子的脂肪酸也将 研究过。 将进行离体肺灌注研究 确定15-HETE的代谢是否可能发生在整个 肺。 肺PGDH的酶活性被诱导20-至 兔子怀孕期间增加了 40 倍，但我们的研究表明 免疫反应蛋白的量没有增加。 一个 肺抑制因子 PGDH 是从肺中分离出来的 怀孕的兔子。 抑制剂随中性脂质迁移 部分，证据表明它由游离脂肪酸组成。 将进一步检查该因素以确定其身份和 确定怀孕期间 PGDH 活性的增加是否与 由于该抑制剂的变化。 这种抑制剂的存在是 对于理解表达和调节很重要 PGD​​H 活性。 PGD​​H 抗体已制成，并将 用于通过免疫细胞化学定位酶 肺部光学和电子显微镜水平的手术 和其他兔组织。 抗肺PGDH也将用于 表征并比较免疫化学特性 来自各种组织的 PGDH 酶。 PGD​​H活性被抑制 香烟烟雾中的物质和酶的损失 活动可能会导致血管和气道的变化 在吸烟者中观察到。 反应活性物质丙烯醛的作用 香烟烟雾中的醛，将研究PGDH活性。 本提案中描述的研究将带来更大的成果 该酶的特性及其功能 肺中的前列腺素和其他类二十烷酸衍生物 功能、妊娠、肺和心血管 疾病。