MECHANISMS OF TOXIC INJURY IN VASCULAR TISSUE

我们的长期目标是阐明血管损伤过程中的细胞事件 有毒化学物质。我们持续关注烯丙胺（AA）代谢， 一种有毒的醛，丙烯醛，可能引起线粒体损伤 (and随后的血管增殖反应）将被增强， 大血管坏死性损伤机制的研究 血管毒性协同作用模型（AA加β-氨基丙腈，或 betaAPN）。 我们将剖析血管平滑肌细胞发生的早期代谢事件， 肌肉细胞，衡量丙烯醛的直接攻击的贡献， 细胞亲核试剂与H2 O2和活性氧的贡献 物种毒性。损伤的潜在作用和机制 建议的目标细胞器，线粒体，将被检查。我们将定义 血管（主动脉， 内皮细胞和中膜）细胞的急性AA暴露 从用AA体内处理的大鼠中。我们将描述早期反应 基因（c-myc、c-fos）、热休克蛋白（HSP 70）和生长的表达 因素我们将定义AA急性诱导的表型变化 （形态学、细胞骨架、平滑肌α-肌动蛋白表达），以及 确定AA代谢抑制剂是否会抑制这些急性 应答 为了更好地理解协同毒性，我们将进行一系列的 在培养的血管平滑肌细胞中的体外机制实验， 他们的媒体，和重组酶系统，以评估的重要性 SSAO在血管损伤中的作用，并评估丙烯醛的重要性， 谷胱甘肽结合物、加合物和其他毒性代谢物 其被提出通过靶向线粒体来启动。 这些研究将有希望导致对血管的理解 代谢、损伤和病理反应。

Our long-range goal is to clarify cellular events during vascular injury by toxic chemicals. Our continued focus on allylamine (AA)-metabolism to a toxic aldehyde, acrolein, that causes probable mitochondrial injury (and a later vascular proliferative response) will be augmented by studies of the mechanism of large vessel necrotizing injury in a new model of vascular toxic synergism (AA plus beta-aminopriopionitrile, or betaAPN). We will dissect early metabolic events occurring in vascular smooth muscle cells, weighing the contribution of acrolein's direct attack on cellular nucleophiles vs. the contribution of H2O2 and reactive oxygen species to toxicity. The potential role and mechanisms of injury to the proposed target organelle, mitochondria, will be examined. We will define altered growth and cell division characteristics of vascular (aortic, endothelial and medial) cells due to acute AA exposure in cells derived from rats treated in vivo with AA. We will characterize early response genes (c-myc, c-fos) heat shock proteins (HSP70) and expression of growth factors. We will define phenotypic changes induced acutely by AA (morphology, cytoskeleton, smooth muscle alpha-actin expression), and ascertain if inhibitors of AA metabolism will obviate these acute responses. To better understand synergistic toxicity, we will do a series of in vitro mechanistic experiments in cultured vascular smoothmuscle cells, their media, and reconstituted enzyme systems to assess the importance of SSAO in vascular damage, and assess the importance of acrolein, glutathione conjugates, adducts, and other toxic metabolites in damage which is proposed to be initiated through targeting of mitochondria. These studies will hopefully lead to an understanding of vascular metabolism, injury, and pathologic responses.