CARBON MONOXIDE AND VASCULAR SMOOTH MUSCLE CELL FUNCTION

这项研究提案的广泛、长期目标是建立 血管平滑肌细胞来源的一氧化碳(CO)作为一种新的 和生物上重要的信使分子，调节血管 血管壁损伤部位的动态平衡。我们最近发现 SMC通过血红素加氧酶-1(HO-1)的分解代谢产生一氧化碳 特定的体液和血液动力学因素在 血管损伤微环境诱导HO-1基因表达与一氧化碳 放手。基于这些发现，我们建议研究生物 一氧化碳对血管SMC功能的影响这些研究将确定 外源性和内源性CO对血管紧张素转换酶的影响 血管张力、SMC增殖、SMC胶原合成。总而言之， 显示CO改变SMC功能的情况下，我们将确定 这种效应的分子基础。因为我们已经证明了钴能刺激 血管SMC中可溶性鸟苷环化酶的活性，我们将进行研究 CGMP和蛋白激酶G(PKG)和/或A(PKA)是否介导 CO的生物效应。我们将进一步探讨CO作为一种 通过产生过度表达的SMC来实现SMC功能的自分泌调节 HO-1酶。如果HO-1转染的SMC改变了我们的生物学反应 将尝试通过使用特定的SMC来恢复响应 金属原卟啉HO抑制剂或HO-1反义治疗SMC 寡聚脱氧核苷酸。最后，我们将研究一氧化碳在 大鼠颈动脉内膜增厚和血管反应性的调节 动脉球囊损伤模型。我们将确定当地是否 抑制或过度表达HO-1活性和CO产生调节 血管损伤部位的病变形成和血管张力。在……里面 此外，我们将确定cGMP和PKG/PKA是否负责 CO在体内的任何作用。预计这些研究 将(A)建立CO作为一种新的SMC衍生的信号分子 调节SMC收缩、增殖和胶原合成，以及(B) 导致治疗中新的治疗策略的发展 闭塞性血管疾病。

The broad, long-term objective of this research proposal is to establish vascular smooth muscle cell (SMC)-derived carbon monoxide (CO) as a novel and biologically important messenger molecule that regulates vascular homeostasis at sites of vessel wall injury. We have recently discovered that SMC generate CO from the catabolism of heme oxygenase-1 (HO-1) and that specific humoral and hemodynamic factors encountered in the microenvironment of vascular injury induces HO-1 gene expression and CO release. Based on these findings, we propose to study the biological effects of CO on vascular SMC function. These studies will determine the effect of both exogenously administered and endogenously-derived CO on blood vessel tone, SMC proliferation, and SMC collagen synthesis. In all instances where CO is shown to alter SMC function we will determine the molecular basis of this effect. Since we have shown that Co stimulates soluble guanylate cyclase activity in vascular SMC, we will investigate whether cGMP and protein kinase G (PKG) and/or A (PKA) mediates the biological effects of CO. We will further explore the role of CO as an autocrine regulator of SMC function by generating SMC that over-express HO-1 enzyme. If HO-1 transfected SMC have altered biological responses we will attempt to restore responses by treating SMC with specific metalloprotoporphyrin HO inhibitors or by treating SMC with HO-1 antisense oligodeoxynucleotides. Finally, we will investigate the role of CO in regulating intimal thickening and vascular reactivity in the rat carotid artery balloon injury model. We will determine whether the local inhibition or over-expression of HO-1 activity and CO production regulates lesion formation and vascular tone at sites of vascular injury. In addition, we will determine whether cGMP and PKG/PKA are responsible for any of the in vivo effects of CO. It is anticipated that these studies will (a) establish CO as a novel SMC-derived signaling molecule that regulates SMC contraction, proliferation and collagen synthesis, and (b) lead to the development of novel therapeutic strategies in treating occlusive vascular disease.