NO-induced vascular smooth muscle cell motility

NO诱导血管平滑肌细胞运动

• 批准号: 7163460
• 负责人: AVIV HASSID
• 金额: $ 35.44万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7163460
• 关键词: 1-Phosphatidylinositol 3-Kinase; AGTR2 gene; Abbreviations; Accounting; Alleles; Analysis of Variance; Angiotensin II; Angiotensin Receptor; Antigens; Arginine; Atherosclerosis; Attenuated; Blood Vessels; Bromodeoxyuridine; C-Type Natriuretic Peptide; Cell membrane; Chronic; Condition; Copy Number Polymorphism; Cultured Cells; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; DNA biosynthesis; DNA chemical synthesis; Elevation; Event; Extracellular Signal Regulated Kinases; Glycine; Hemagglutinin; Hyperinsulinism; Injury; Insulin; Insulin Resistance; Internal Ribosome Entry Site; Investigation; LY294002; Letters; Mediating; Metabolic syndrome; Mitogen-Activated Protein Kinases; Modeling; Mus; N-acetylpenicillamine; Neuro-Oncological Ventral Antigen 2; Nitric Oxide; Non-Insulin-Dependent Diabetes Mellitus; Obesity; PH Domain; PTPN11 gene; Pathogenesis; Pathology; Phenylmethylsulfonyl Fluoride; Phosphatidylinositols; Platelet-Derived Growth Factor; Play; Polymerase Chain Reaction; Prevalence; Protein Tyrosine Phosphatase; Proteins; Publications; Purpose; Quinoxalines; Rate; Rattus; Recruitment Activity; Reporting; Research Personnel; Role; S-nitro-N-acetylpenicillamine; Saralasin; Smooth Muscle Myocytes; Sodium Dodecyl Sulfate-PAGE; Testing; Tyrosine; Vascular Diseases; adapter protein; cell motility; enhanced green fluorescent protein; human NOS2A protein; human NOS3 protein; inhibitor/antagonist; insight; kinase inhibitor; neointima formation; programs; protein aminoacid sequence; receptor; research study; response; vascular smooth muscle cell proliferation

DESCRIPTION (provided by applicant): Nitric oxide (NO) is generally considered to play a protective role in blood vessels. However, others and we have obtained evidence that this may not always be the case but the mechanisms underlying diverse responses to NO are not known. The purpose of this proposal is to test an exciting new hypothesis on the capacity of chronically elevated insulin levels to switch the role of nitric oxide from protective to deleterious substance in blood vessels. We have found that the inhibitory effects of NO on both motility and proliferation are abrogated in vascular smooth muscle cells chronically treated with insulin. These findings support a new hypothesis on the role of insulin as a switcher of vascular smooth muscle cell phenotypic responses to NO. Our preliminary results indicate that the motility-stimulatory effect of NO, uncovered by chronic insulin treatment of cultured rat aortic smooth muscle cells, is associated with increased PI 3 kinase activity and requires the functional availability of angiotensin II, of the adapter protein Gab1 and the protein tyrosine phosphatase SHP2. Studies by others have found that Gab1 can be recruited to the plasma membrane via increased PIPS levels. However, the mechanistic linkage of chronic insulin treatment to Gab1 and SHP2 function has not been defined. Moreover, experiments to determine whether similar mechanisms may be applicable to abrogation of the antiproliferative effect of NO by chronic insulin treatment have not been performed. Finally, the pathophysiological significance of our results is unknown. We propose to implement the following specific aims: Aim 1: To determine whether increased angiotensin II function is necessary and/or sufficient to account for the effect of insulin on PI3K activity and NO-induced cell motility. Aim 2: To determine whether chronic insulin treatment recruits Gab1 to the cell membrane and whether insulin- independent recruitment of Gab1 or SHP2 to the cell membrane can mimic the motility-stimulatory effect of NO uncovered by chronic insulin treatment. Aim 3: To uncover mechanisms that describe how hyperinsulinemia attenuates the effect of NO as inhibitor of PDGF-induced DNA synthesis, in cultured rat aortic smooth muscle cells. Aim 4: To determine whether expression of inducible nitric oxide synthase in vascular injury enhances neointima formation in hyperinsulinemic mice, but has the opposite effect in normoinsulinemic mice or in hyperinsulinemic mice treated with an AT1 receptor antagonist.

描述（由申请人提供）：一氧化氮（NO）通常被认为在血管中起保护作用。然而，其他人和我们已经获得的证据表明，这可能并不总是如此，但对NO的不同反应的机制尚不清楚。该提案的目的是测试一个令人兴奋的新假设，即长期升高的胰岛素水平将一氧化氮的作用从血管中的保护物质转变为有害物质的能力。我们已经发现，在血管平滑肌细胞长期用胰岛素处理，NO对运动和增殖的抑制作用被废除。这些发现支持了一个新的假说，即胰岛素是血管平滑肌细胞对NO表型反应的转换器。我们的初步结果表明，NO的运动刺激作用，通过培养的大鼠主动脉平滑肌细胞的慢性胰岛素治疗发现，与PI 3激酶活性增加有关，并需要血管紧张素II的功能可用性，衔接蛋白Gab 1和蛋白质酪氨酸磷酸酶SHP 2的结合。其他人的研究发现，Gab 1可以通过增加PIPS水平被招募到质膜。然而，长期胰岛素治疗与Gab 1和SHP 2功能的机制联系尚未确定。此外，还没有进行实验来确定类似的机制是否可适用于通过长期胰岛素治疗来消除NO的抗增殖作用。最后，我们的研究结果的病理生理意义是未知的。我们建议实现以下具体目标：目的1：确定是否增加血管紧张素II功能是必要的和/或足以解释胰岛素对PI 3 K活性和NO诱导的细胞运动的影响。目标二：确定慢性胰岛素治疗是否将Gab 1募集到细胞膜，以及Gab 1或SHP 2的胰岛素非依赖性募集到细胞膜是否可以模拟慢性胰岛素治疗所揭示的NO的运动刺激作用。目标3：揭示高胰岛素血症如何减弱NO作为PDGF诱导的DNA合成抑制剂的作用的机制。目标4：确定诱导型一氧化氮合酶在血管损伤中的表达是否增强高胰岛素血症小鼠的新生内膜形成，但在正常胰岛素血症小鼠或用AT 1受体拮抗剂治疗的高胰岛素血症小鼠中具有相反的作用。