CARBON MONOXIDE AND VASCULAR SMOOTH MUSCLE CELL FUNCTION

一氧化碳与血管平滑肌细胞功能

• 批准号: 7025793
• 负责人: WILLIAM DURANTE
• 金额: $ 25.12万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7025793
• 关键词: autocrine; cGMP dependent protein kinase; carbon monoxide; cardiovascular injury; cell communication molecule; cell growth regulation; cell proliferation; collagen disorder; cyclic GMP; genetically modified animals; heme oxygenase; laboratory mouse; laboratory rat; muscle function; nonhuman therapy evaluation; paracrine; respiratory therapy; tissue /cell culture; vascular endothelial growth factors; vascular smooth muscle

DESCRIPTION (provided by applicant): The broad long-term objective of this research proposal is to establish heme oxygenase-1 (HO-1)-derived carbon monoxide (CO) as a novel and biologically important gas that regulates homeostasis at sites of vascular injury. We have measured the release of CO from vascular smooth muscle cells (SMC) and found that SMC-derived CO functions in an autocrine and paracrine fashion to inhibit SMC proliferation and platelet aggregation, respectively. The central hypothesis of this proposal is that HO-1-derived CO is a critical regulator of the SMC response to vascular injury. To test our hypothesis we plan to pursue the following three complementary and linked specific aims. In aim 1, we will examine the role of CO in regulating vascular SMC migration, collagen synthesis, and the secretion of vascular endothelial growth factor (VEGF) utilizing cultured vascular SMC. The effect of exogenously administered and endogenously derived CO will be studied. SMC will be exposed to CO via an exposure chamber while endogenous CO production will be induced by adenovirus-mediated transfer of the HO-1gene. The role of HO-1-derived CO in regulating SMC function will also be examined by harvesting SMC from the aorta of HO-1 knockout animals and comparing their functional properties with SMC from wild type animals. If CO is found to alter these SMC functions, we will determine the involvement of the cGMP or p38 mitogen activated protein kinase signaling pathways. In aim 2, we will elucidate the actions of HO-1 in regulating collagen deposition and VEGF expression following arterial injury using transgenic mice deficient in HO-1. In addition, we will investigate if CO inhalation can substitute for HO-1 in preventing collagen deposition and VEGF expression in these animals. In aim 3, we will explore the effect of adenovirus-mediated HO-1 gene delivery on collagen accumulation and VEGF expression in these animals. Finally, we will determine if CO-mediated VEGF release functions in a paracrine manner to stimulate endothelial cell growth both in vitro and in vivo. It is anticipated that these studies will (a) establish CO as a novel regulator of the vessel wall's response to injury and (b) implicate the HO-1/CO system as a promising new therapeutic target in treating vascular fibroproliferative disease.

描述（由申请人提供）：本研究计划的广泛长期目标是将血红素加氧酶-1 (HO-1) 衍生的一氧化碳 (CO) 确立为一种新型且具有生物学重要意义的气体，可调节血管损伤部位的稳态。我们测量了血管平滑肌细胞（SMC）释放的CO，发现SMC衍生的CO以自分泌和旁分泌的方式分别抑制SMC增殖和血小板聚集。该提议的中心假设是 HO-1 衍生的 CO 是 SMC 对血管损伤反应的关键调节剂。为了检验我们的假设，我们计划追求以下三个互补且相互关联的具体目标。在目标 1 中，我们将利用培养的血管 SMC 检查 CO 在调节血管 SMC 迁移、胶原蛋白合成和血管内皮生长因子 (VEGF) 分泌中的作用。将研究外源性施用和内源性 CO 的影响。 SMC 将通过暴露室暴露于 CO，而内源性 CO 的产生将由腺病毒介导的 HO-1 基因转移诱导。 HO-1 衍生的 CO 在调节 SMC 功能中的作用也将通过从 HO-1 敲除动物的主动脉收获 SMC 并将其功能特性与来自野生型动物的 SMC 进行比较来检查。如果发现 CO 改变这些 SMC 功能，我们将确定 cGMP 或 p38 丝裂原激活蛋白激酶信号通路的参与。在目标 2 中，我们将使用 HO-1 缺陷的转基因小鼠阐明 HO-1 在动脉损伤后调节胶原蛋白沉积和 VEGF 表达的作用。此外，我们将研究 CO 吸入是否可以替代 HO-1 来预防这些动物中的胶原蛋白沉积和 VEGF 表达。在目标 3 中，我们将探讨腺病毒介导的 HO-1 基因递送对这些动物中胶原蛋白积累和 VEGF 表达的影响。最后，我们将确定 CO 介导的 VEGF 释放是否以旁分泌方式发挥作用，以刺激体内和体外的内皮细胞生长。预计这些研究将（a）确立 CO 作为血管壁损伤反应的新型调节剂，（b）表明 HO-1/CO 系统作为治疗血管纤维增殖性疾病的有前途的新治疗靶点。