CYTOKINE REGULATION OF ENDOTHELIUM PROSTANOID SYNTHESIS

肿瘤等细胞因子对内皮细胞功能的调控 坏死因子和白细胞介素1（IL.1）* 用于调节炎症 和组织修复。这些细胞因子是众所周知的花生四烯酸的诱导剂。 酸代谢，在EC中，它们引起前列环素的合成 （PGI-2）。PGI-2与其他信号结合，是一种有效的血管扩张剂 促进白细胞聚集和外渗。的几名成员 负责PGI-2合成的细胞内酶级联反应已经 最近被克隆和鉴定，包括胞浆磷脂酶 A2（cPLA 2）和环加氧酶（Cox-l和考克斯-2）。最近的证据 表明cPLA 2和考克斯两者可能具有不同的组成性， 神经调节的同种型，并且在表达中存在变化， 这些酶在几种细胞系统中由IL-1和TNF诱导，包括 EC.然而，受调节的精确异构体和 在细胞因子处理EC中发生的酶活性不清楚 明白在此期间，我将进行系统的 方法来了解这些调节的分子方面 人脐静脉EC中TNF和IL-1的酶，包括分析 mRNA转录率、mRNA和蛋白质稳定性和水平，以及 酶活性的变化。将使用的技术包括核 径流，北方印迹，蛋白质印迹，免疫沉淀和 生化酶测定这些研究将提供一个完整的 描述了细胞因子调节这一关键的生物学途径， EC.

The control of endothelial cell (EC) function by cytokines such as tumor necrosis factor and interleukin 1 (IL.1)* serves to regulate inflammation and tissue repair. These cytokines are well known inducers of arachidonic acid metabolism, and in EC they bring about synthesis of prostacyclin (PGI-2). PGI-2 in combination with other signals, is a potent vasodilator that promotes leukocyte recruitment and extravasation. Several members of the intracellular, enzymatic cascade responsible for PGI-2 synthesis have recently been cloned and characterized, including cytosolic phospholipase A2 (cPLA2) and cyclooxygenases (Cox-l and Cox-2). Recent evidence indicates that both cPLA2 and Cox may have distinct constitutive and hormonally regulated isoforms, and that there are changes in expression these enzymes induced by IL-I and TNF in several cell systems including EC. However, the precise isoforms which are regulated and the changes in enzymatic activity that occur in cytokine treated EC are not clearly understood. During the period of this grant, I will pursue a systematic approach to understand the molecular aspects of regulation of these enzymes by TNF and IL-1 in human umbilical vein EC, including analysis of mRNA transcriptional rates, mRNA and protein stability and levels, and changes in enzyme activities. Techniques to be utilized include nuclear runoff, northern blotting, western blotting, immunoprecipitation and biochemical enzyme assays. These studies should provide a complete description of cytokine regulation of this critical biological pathway in EC.