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：
7333236
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：
2000-08-01 至 2009-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7333236
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

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 Gab1and 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.

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，但背后的机制多种多样对 NO 的反应尚不清楚。该提案的目的是测试一个令人兴奋的新假设长期升高的胰岛素水平将一氧化氮的作用从保护作用转变为有害作用的能力血管中的物质。我们发现NO对运动和增殖都有抑制作用在长期接受胰岛素治疗的血管平滑肌细胞中会被消除。这些发现支持了一个新的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假设。我们的初步结果表明，慢性胰岛素所揭示的 NO 的运动刺激作用对培养的大鼠主动脉平滑肌细胞进行治疗，与 PI 3 激酶活性增加有关需要血管紧张素 II、衔接蛋白 Gab1 和蛋白酪氨酸的功能可用性磷酸酶SHP2。其他人的研究发现 Gab1 可以通过以下途径被招募到质膜上：增加 PIPS 水平。然而，长期胰岛素治疗与 Gab1 和 SHP2 的机制联系函数尚未定义。此外，还需要进行实验来确定是否可以使用类似的机制适用于通过长期胰岛素治疗消除 NO 的抗增殖作用尚未得到证实执行。最后，我们的结果的病理生理学意义尚不清楚。我们建议实施具体目标如下：目标 1. 确定是否有必要增加血管紧张素 II 功能和/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. 至确定长期胰岛素治疗是否会将 Gab1 募集至细胞膜以及胰岛素是否 Gab1 或 SHP2 独立募集到细胞膜上可以模拟运动刺激作用长期胰岛素治疗未发现NO。目标 3：揭示描述如何进行的机制在培养大鼠中，高胰岛素血症减弱 NO 作为 PDGF 诱导的 DNA 合成抑制剂的作用主动脉平滑肌细胞。目的 4. 确定诱导型一氧化氮合酶是否在血管损伤增强高胰岛素血症小鼠的新内膜形成，但在正常胰岛素血症小鼠或用 AT1 受体拮抗剂治疗的高胰岛素血症小鼠。