Cytochrome P450 regulation by hyperoxia and nitric oxide

高氧和一氧化氮对细胞色素 P450 的调节

• 批准号: 6549740
• 负责人: XANTHI Ioanna COUROUCLI
• 金额: $ 10.01万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6549740
• 关键词: aromatic hydrocarbon receptor; cytochrome P450; disease /disorder prevention /control; fluorimetry; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rat; lung injury; medical complication; newborn animals; nitric oxide; oxygen therapy; polymerase chain reaction; respiratory disease /disorder therapy; respiratory insufficiency /failure; respiratory therapy; western blottings

DESCRIPTION (provided by applicant): Inhaled nitric oxide (INO) and supplemental oxygen are used clinically in the treatment of pulmonary diseases associated with respiratory failure in infants. In preterm and term human infants, oxygen and/or nitric oxide therapy may contribute to lung injury. The long-term goal of this application is to develop rational strategies for the prevention/treatment of lung pathologies in infants exposed to hyperoxia and INO. This application comprises 4 Specific Aims: (1) To characterize the specific mechanisms by which hyperoxia and INO, alone or in combination, modulate hepatic and pulmonary cytochrome P450 (CYP) enzyme expression, in relation to acute lung injury. Newborn rats will be exposed to hyperoxia (geater than 95 percent oxygen), INO, a mixture of hyperoxia and INO, or maintained in room air for selected time points for up to 7 days. Message levels [(Northern hybridization, reverse transcriptase polymerase chain reaction (RT-PCR), in situ hybridization), apoprotein contents (Western blotting and immunohistochemistry), and enzyme activities (fluorimetry) of specific CYP isoforms, i.e. CY`PlAl/lA2 and CYP2EI will be determined in the liver and lung. (2) To test the hypothesis that endogenous NO contributes to the downregulation of specific CYP enzymes by hyperoxia, and to the mechanism(s) of lung injury by hyperoxia. Mice lacking the gene for NOS2 or NOS3 will be used to specifically test the hypothesis that endogenous NO contributes to hyperoxic lung injury. (3) To determine the possible role of the aryl hydrocarbon receptor (AHR) in the differential susceptibilities of newborn rodents exposed to hyperoxia or hyperoxia + INO. AHR null mice will be used to determine the possible role of the AHR in acute lung injury. (4) To test the hypothesis that neonatal imprinting of CYP enzymes caused by exposure to hyperoxia, [NO, or hyperoxia + INO contributes to abnormal lung maturation in adult rats. The results will provide critical information about the acute effects of INO and hyperoxia on CYP expression, and the contribution of neonatal imprinting of these enzymes in abnormal lung maturation occurring through adult life. The studies will also help identify appropriate supportive therapies to minimize any adverse effects arising out of this clinically important therapy.

描述（由申请人提供）： 吸入一氧化氮（INO）和补充氧气在临床上用于 与呼吸衰竭相关的肺部疾病的治疗 婴儿。 在早产和足月人类婴儿中，氧气和/或一氧化氮 治疗可能导致肺损伤。 这个应用程序的长期目标是 制定合理的策略来预防/治疗肺部疾病 暴露于高氧和INO的婴儿的病理学。 本申请 包括4个具体目标：（1）通过以下方式表征具体机制： 其中高氧和INO单独或联合调节肝脏和 肺细胞色素P450（CYP）酶表达，与急性肺 损伤 新生大鼠将暴露于高氧（高于95%） 氧），INO，高氧和INO的混合物，或保持在室内空气中， 选定的时间点长达7天。 消息级别[（北方 杂交，逆转录聚合酶链反应（RT-PCR）， 原位杂交），载脂蛋白含量（Western印迹和 免疫组织化学）和酶活性（荧光测定法）的特异性 将在肝脏中测定同工型，即CYP 1A 1/1A 2和CYP 2 E1， 肺。(2)为了验证内源性NO参与细胞凋亡的假设， 通过高氧下调特异性β-内酰胺酶，及其机制 高氧肺损伤 缺乏NOS 2或NOS 3基因的小鼠， 用于专门测试内源性NO有助于 高氧肺损伤(3)确定芳基的可能作用 碳氢化合物受体与新生儿的分化潜能 暴露于高氧或高氧+ INO的啮齿动物。 将使用AHR敲除小鼠 以确定AHR在急性肺损伤中的可能作用。(4)测试 这一假说认为，新生儿印迹的酶所造成的接触， 高氧、[NO]或高氧+ INO可导致肺成熟异常， 成年大鼠 结果将提供关于急性 INO和高氧对p53表达的影响，以及 这些酶的新生儿印迹在异常肺成熟发生 通过成人生活。 这些研究还将有助于确定适当的 支持治疗，以尽量减少由此产生的任何不良反应 重要的临床治疗。