NO-induced vascular smooth muscle cell motility

NO诱导血管平滑肌细胞运动

基本信息

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
项目类别：
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

项目摘要

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）通常被认为对血管具有保护作用。然而，其他人和我们已经获得的证据表明情况可能并不总是如此，但对 NO 的不同反应背后的机制尚不清楚。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能否将血管中一氧化氮的作用从保护性物质转变为有害物质。我们发现，在长期接受胰岛素治疗的血管平滑肌细胞中，NO 对运动和增殖的抑制作用被消除。这些发现支持了关于胰岛素作为血管平滑肌细胞对 NO 表型反应的转换者的作用的新假设。我们的初步结果表明，通过长期胰岛素治疗培养的大鼠主动脉平滑肌细胞发现，NO 的运动刺激作用与 PI 3 激酶活性增加有关，并且需要血管紧张素 II、衔接蛋白 Gab1 和蛋白酪氨酸磷酸酶SHP2。其他人的研究发现，Gab1 可以通过增加 PIPS 水平而被招募到质膜上。然而，长期胰岛素治疗与 Gab1 和 SHP2 功能的机制联系尚未明确。此外，还没有进行实验来确定类似的机制是否适用于通过长期胰岛素治疗消除NO的抗增殖作用。最后，我们的结果的病理生理学意义尚不清楚。我们建议实现以下具体目标：目标 1：确定增加血管紧张素 II 功能是否必要和/或足以解释胰岛素对 PI3K 活性和 NO 诱导的细胞运动的影响。目标 2：确定长期胰岛素治疗是否会将 Gab1 募集至细胞膜，以及 Gab1 或 SHP2 与胰岛素无关地募集至细胞膜是否可以模拟长期胰岛素治疗所发现的 NO 的动力刺激作用。目标 3：在培养的大鼠主动脉平滑肌细胞中，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：确定血管损伤中诱导型一氧化氮合酶的表达是否会增强高胰岛素血症小鼠的新内膜形成，但在正常胰岛素小鼠或用 AT1 受体拮抗剂治疗的高胰岛素血症小鼠中具有相反的作用。