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: 8583336
• 负责人: KAFAIT U MALIK
• 金额: $ 37.85万
• 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/8583336
• 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.

描述（由申请人提供）：当前的提案是我们在上一个赠款期间进行的研究的扩展，目的是研究通过CYP1B1产生的花生四烯酸（AA）代谢产物的贡献，以及参与ANG II诱导的新内膜生长引起的基础机制，由血管造成的脉管造成的疾病和动脉粥样硬化引起。它基于我们新的初步数据：a）选择性CYP1B1抑制剂2,4,32,52-四甲氧基苯二甲酸酯（TMS）或腺病毒CYP1B1 shRNA（CYP1B1 SHRNA）会导致ANG II刺激的新生长，导致Rat Carotid Carotid Carotid carotid carotid artery的新刺激性生长。 b）ANG II增加了野生型颈动脉（CYP1B1+/+）的颈动脉的新内膜生长，但不足CYP1B1缺乏（CYP1B1 - / - ），小鼠； c）用TMS治疗抑制脂肪条纹区域的产生及其在Apo e敲除（APOE - / - ）小鼠中的ANG II加速度的加速度（HFD）； d）ANG II和AA诱导的大鼠VSMC中的NADPH氧化酶活性由TMS或CYP1B1 shRNA预防，而在CYP1B1 - / - 的VSMC中，但不是CYP1B1+/+，而不是小鼠。 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.为了检验这一假设，我们将解决以下特定目的：目标1。确定CYP1B1对气球受损害的大鼠和受伤的小鼠颈动脉刺激的新内膜生长的贡献； AIM 2A）研究CYP1B1对小鼠颈动脉电线损伤后对动脉粥样硬化和新内膜生长的贡献； AIM 2B）检查CYP1B1对ANG II诱导的动脉粥样硬化，动脉瘤和新内膜生长的贡献；目的。 3。研究小鼠高胆固醇血症的VSMC中CYP1B1依赖性ANG II-和AA诱导的NADPH氧化酶的机制。为了解决这些目标，我们计划使用大量的体外和体内细胞和分子生物学技术以及转基因小鼠。其中包括使用1）腺病毒CYP1B1 shRNA； 2）apoe - / - /cyp1b1+/+和双基因敲除 - / - / - /cyp1b1 - / - 以及我们在实验室中生成的apein-apoe+/+/+/+/+/cyp1b1+/+小鼠； 3）从这些转基因动物中分离出VSMC； 4）形态学，组织学，免疫组织化学和荧光以及生化技术，以及5）用于鉴定AA代谢物的HPLC-LC-ESI-MS。拟议的研究应提高我们当前对涉及再狭窄和动脉粥样硬化发病机理的细胞和分子机制的了解，并使我们能够证明CYP1B1是开发更有效的治疗药物（例如TMS，例如治疗ANG II诱导的再狭窄引起的血管损伤和动脉粥样硬化症引起的血管造成的再狭窄）的潜在新颖靶标。