Eicosanoid-Induced Vascular Growth During Injury

受伤期间类二十烷酸诱导的血管生长

• 批准号: 8238737
• 负责人: KAFAIT U MALIK
• 金额: $ 39.92万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8238737
• 关键词: Acceleration; Address; Adenoviruses; Aneurysm; Angiotensin II; Animals; Aortic Injury; Apolipoprotein E; Arachidonic Acids; Area; Arterial Fatty Streak; Arteries; Atherosclerosis; Biochemical; Blood Vessels; CYP1B1 gene; Carotid Arteries; Cells; Cytochrome P450; Data; Development; Diet; Eicosanoids; Fatty acid glycerol esters; Fluorescence; Generations; Grant; Growth; High Pressure Liquid Chromatography; Hypertrophy; In Vitro; Infiltration; Inflammation; Inflammatory; Injury; Knock-out; Knockout Mice; Knowledge; Laboratories; Lesion; Leukocytes; Lipid Peroxides; Lipids; MAPK3 gene; Mass Spectrum Analysis; Measures; Metabolism; Molecular; Molecular Biology Techniques; Mus; NADP; NADPH Oxidase; Oxidases; Pathogenesis; Pharmaceutical Preparations; Plasma; Production; Protein Kinase; Rattus; Reactive Oxygen Species; Research; Research Design; Role; SRC gene; Signaling Molecule; Techniques; Testing; Therapeutic; Transgenic Animals; Transgenic Mice; Vascular Diseases; apolipoprotein E-2; base; feeding; hypercholesterolemia; in vivo; inhibitor/antagonist; injured; macrophage; migration; novel; prevent; restenosis; small hairpin RNA

DESCRIPTION (provided by applicant): The current proposal is an extension of our studies conducted over the previous grant period and is aimed to investigate the contribution of arachidonic acid (AA) metabolites generated via CYP1B1 and the underlying mechanism involved in Ang II-induced neointimal growth caused by vascular injury and atherosclerosis. It is based on our novel preliminary data that: a) the selective CYP1B1 inhibitor 2,4,32,52-tetramethoxystilbene (TMS) or adenovirus CYP1B1 shRNA (CYP1B1 shRNA) prevents Ang II-stimulated neointimal growth caused by balloon injury of rat carotid artery; b) Ang II increases neointimal growth in wire-injured carotid artery of wild type (Cyp1b1+/+),but not CYP1B1 deficient (Cyp1b1-/-), mice; c) treatment with TMS inhibits generation of fatty streak areas and their acceleration by Ang II in Apo E knockout (apoE-/-) mice fed a high fat diet (HFD); and d) Ang II and AA-induced NADPH oxidase activation in rat VSMCs is prevented by TMS or CYP1B1 shRNA, and in VSMC from Cyp1b1-/-, but not Cyp1b1+/+, mice. These findings have led us to the following central hypothesis: CYP1B1, through AA metabolites/lipid peroxides, results in activation of NADPH oxidase and production of ROS, which, by activating one or more signaling molecules, contribute to the pathogenesis of restenosis caused by Ang II during vascular injury and to atherosclerosis produced by hypercholesterolemia and its acceleration by Ang II. To test this hypothesis, we will address the following specific aims: Aim 1. Determine the contribution of CYP1B1 to neointimal growth stimulated by Ang II in balloon-injured rat and wire- injured mouse carotid artery; Aim 2a) Investigate the contribution of CYP1B1 to atherosclerosis and neointimal growth after wire injury of mouse carotid artery; Aim 2b) Examine the contribution of CYP1B1 to Ang II-induced acceleration of atherosclerosis, aneurysm, and neointimal growth; Aim. 3. Investigate the mechanism of CYP1B1-dependent Ang II- and AA-induced NADPH oxidase and ROS production in VSMCs from hypercholesterolemia in mice. To address these aims, we plan to use numerous state-of-the-art in vitro and in vivo cellular and molecular biology techniques and transgenic mice. These include using 1) adenovirus CYP1B1 shRNA; 2) apoE-/-/Cyp1b1+/+ and double knockout-apoE-/-/Cyp1b1-/- and knockin-apoE+/+/CYp1b1+/+ mice that we have generated in our laboratory; 3) isolated VSMCs from these transgenic animals; 4) morphological, histological, immunohistochemical and fluorescence, and biochemical techniques, and 5) HPLC-LC-ESI-MS for the identification of AA metabolites. The proposed studies should advance our current knowledge of the cellular and molecular mechanisms involved in the pathogenesis of restenosis and atherosclerosis and allow us to demonstrate CYP1B1 as a potential novel target for the development of more effective therapeutic drugs such as TMS for treating Ang II-induced restenosis caused by vascular injury and atherosclerosis by hypercholesterolemia. PUBLIC HEALTH RELEVANCE: The proposed studies are designed to investigate the contribution of cytochrome P450 1B1 to the action of angiotensin II to promote restenosis caused by vascular injury and aortic atherosclerotic lesions and aneurysms produced by hypercholesterolemia. They should advance our current knowledge of the mechanisms involved in the pathogenesis of this vascular disease. Moreover, they should allow us to demonstrate cytochrome P450 1B1 as a potential novel target for the development of more effective drugs such as tetramethoxystilbene for treating the pathogenesis of restenosis caused by vascular injury and atherosclerosis.

描述（由申请人提供）：当前的提案是我们在上一个资助期进行的研究的扩展，旨在研究通过CYP 1B 1产生的花生四烯酸（AA）代谢物的贡献以及血管损伤和动脉粥样硬化引起的Ang II诱导的新生内膜生长的潜在机制。基于我们新的初步数据，a）选择性CYP 1B 1抑制剂2，4，32，52-四甲氧基芪（TMS）或腺病毒CYP 1B 1 shRNA CYP 1B 1 shRNA抑制血管紧张素II诱导的大鼠颈动脉球囊损伤后内膜增生; B）血管紧张素II增加野生型钢丝损伤的颈动脉中的新生内膜生长（Cyp 1b 1 +/+），但非CYP 1B 1缺陷c）用TMS处理抑制饲喂高脂饮食（HFD）的Apo E敲除（apoE-/-）小鼠中脂肪条纹区域的产生及其被Ang II加速;和d）TMS或CYP 1B 1 shRNA阻止大鼠VSMC中Ang II和AA诱导的NADPH氧化酶活化，并且在来自Cyp 1b 1-/-但不是Cyp 1b 1 +/+小鼠的VSMC中。这些发现使我们得出以下中心假设：CYP 1B 1通过AA代谢物/脂质过氧化物导致NADPH氧化酶的激活和ROS的产生，其通过激活一种或多种信号分子，促成血管损伤期间由Ang II引起的再狭窄的发病机制和由高胆固醇血症产生的动脉粥样硬化及其由Ang II加速。为了验证这一假设，我们将讨论以下具体目标：目标1。确定CYP 1B 1对球囊损伤的大鼠和线损伤的小鼠颈动脉中由Ang II刺激的新生内膜生长的贡献;目的2a）研究CYP 1B 1对小鼠颈动脉的线损伤后的动脉粥样硬化和新生内膜生长的贡献;目的2b）检查CYP 1B 1对Ang II诱导的动脉粥样硬化、动脉瘤和新生内膜生长的加速的贡献;目的2c）研究CYP 1B 1对血管紧张素II诱导的动脉粥样硬化、动脉瘤和新生内膜生长的加速的贡献。3.探讨CYP 1B 1依赖性血管紧张素II和AA诱导高胆固醇血症小鼠血管平滑肌细胞产生NADPH氧化酶和ROS的机制。为了实现这些目标，我们计划使用许多最先进的体外和体内细胞和分子生物学技术和转基因小鼠。这些方法包括1）腺病毒CYP 1B 1 shRNA; 2）我们在实验室中产生的apoE-/-/Cyp 1b 1 +/+和双敲除apoE-/-/Cyp 1b 1-/-和敲除apoE +/+/CYp 1b 1 +/+小鼠; 3）从这些转基因动物中分离的VSMC; 4）形态学、组织学、免疫组织化学、荧光和生物化学技术; 5）HPLC-LC-ESI-MS鉴定AA代谢产物。拟议的研究应推进我们目前的知识，再狭窄和动脉粥样硬化的发病机制，并使我们能够证明CYP 1B 1作为一个潜在的新的目标，为开发更有效的治疗药物，如TMS治疗血管紧张素II诱导的血管损伤引起的再狭窄和动脉粥样硬化高胆固醇血症。 公共卫生关系：拟定的研究旨在研究细胞色素P450 1B 1对血管紧张素II促进血管损伤和高胆固醇血症引起的主动脉粥样硬化病变和动脉瘤引起的再狭窄的作用。他们应该推进我们目前的知识，在这种血管疾病的发病机制。此外，他们应该使我们能够证明细胞色素P450 1B 1作为一个潜在的新的目标，为发展更有效的药物，如四甲氧基芪治疗血管损伤和动脉粥样硬化引起的再狭窄的发病机制。