Nicotinic receptor effect of ACh on skin endothelium

乙酰胆碱对皮肤内皮的烟碱受体作用

• 批准号: 6756245
• 负责人: KENT T KEYSER
• 金额: $ 3.63万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6756245
• 关键词: acetylcholine; angiogenesis; animal tissue; biological signal transduction; human tissue; immunocytochemistry; neurotransmitter agonist; neurotransmitter antagonist; nicotinic receptors; polymerase chain reaction; receptor binding; receptor expression; skin; skin disorder; vascular endothelium; western blottings

DESCRIPTION (provided by applicant): Nicotinic acetylcholine receptors (nAChRs) are widely expressed in neuronal and non-neuronal tissues. In non-neuronal tissue, nAChRs have been implicated in wound healing, cell proliferation and angiogenesis. Although a few nAChR subunits have been identified in non-neuronal tissues, the combinations of subunits that combine to form functional nAChRs have yet to be determined. In addition, previous experiments that addressed nAChRs in the context of angiogenesis relied upon agonists and antagonists that do not discriminate between different receptor subtypes. We hypothesize that the ACh binding to nAChRs activates intracellular signaling pathways that can trigger angiogenesis. This could have relevance to cutaneous vascular malformations that occur in children. Thus, the goals of this research are to investigate the overall role of nAChRs in angiogenesis, to identify the contributions of specific receptor subtypes, and ultimately to understand the mechanisms whereby activation of nAChRs affects angiogenesis. The Specific Aims are: 1) To determine the effects of cholinergic agonists and antagonists on de novo angiogenesis in normal tissue. This will be evaluated by microvascular sprouting in an ex vivo vascular explant system. 2) To analyze subunit expression in normal vascular endothelial cells and in dermal microvascular endothelial cells. Subaim 1. RT-PCR studies that rely on tested primers sets will be used to determine the expression pattern of nAChR subunits in vascular endothelial cells from normal mice, in the human vascular endothelial cell (HUVEC) in vitro model, and in the human dermal derived microvascular endothelial cells (HDMEC). Subaim 2. To determine the receptor composition of nAChRs expressed in normal vascular tissue. Although a3, a7, and a9 nAChR subunits have been identified in human aortic endothelial cells, their expression on HUVEC and HDMEC as well as the expression of other subunits have not yet been identified and the combinations of subunits that comprise functional nAChRs have yet to be determined. 3) Finally we will evaluate changes in the expression of CD40, VAMC-1, and E-selectin by HUVECs and HDMECs following the administration of cholinergic agonists. The results could have important implications for the pathogenesis and management of proliferative vascular lesions that occur in the skin such as hemangiomas and in inflammatory vascular lesions such as small and medium vessel vasculitis.

描述（由申请人提供）：烟碱乙酰胆碱受体（nAChR）在神经元和非神经元组织中广泛表达。在非神经元组织中，nAChR 与伤口愈合、细胞增殖和血管生成有关。尽管在非神经元组织中已鉴定出一些 nAChR 亚基，但组合形成功能性 nAChR 的亚基组合尚未确定。此外，之前在血管生成背景下研究 nAChR 的实验依赖于不区分不同受体亚型的激动剂和拮抗剂。我们假设 ACh 与 nAChR 结合激活细胞内信号通路，从而触发血管生成。这可能与儿童发生的皮肤血管畸形有关。因此，本研究的目标是研究 nAChR 在血管生成中的总体作用，以确定 特定受体亚型的贡献，并最终了解 nAChR 激活影响血管生成的机制。具体目标是： 1) 确定胆碱能激动剂和拮抗剂对正常组织从头血管生成的影响。这将通过离体血管外植体系统中的微血管发芽来评估。 2)分析正常血管内皮细胞和真皮微血管内皮细胞中的亚基表达。 Subaim 1.依赖于测试引物组的RT-PCR研究将用于确定nAChR亚基在正常小鼠血管内皮细胞、人血管内皮细胞（HUVEC）体外模型和人真皮中的表达模式衍生的微血管内皮细胞（HDMEC）。 Subaim 2. 确定受体 正常血管组织中表达的 nAChR 的组成。尽管a3、a7和a9 nAChR亚基已在人主动脉内皮细胞中被鉴定，但它们在HUVEC和HDMEC上的表达以及其他亚基的表达尚未被鉴定，并且组成功能性nAChR的亚基组合仍有待确定。决定。 3)最后我们将评估给予胆碱能激动剂后HUVEC和HDMEC的CD40、VAMC-1和E-选择素表达的变化。这些结果可能对皮肤中发生的增殖性血管病变（如血管瘤）和炎症性血管病变（如中小血管血管炎）的发病机制和治疗具有重要意义。