CYTOKINE REGULATION OF ENDOTHELIUM PROSTANOID SYNTHESIS

肿瘤等细胞因子对内皮细胞功能的调控 坏死因子和白介素1(IL1)*调节炎症 和组织修复。这些细胞因子是花生四烯酸的众所周知的诱导剂 酸代谢，在EC中它们导致前列环素的合成 (PGI-2)。PGI-2与其他信号相结合，是一种有效的血管扩张剂 这会促进白细胞的募集和外渗。的几位成员 负责PGI-2合成的细胞内，酶的级联反应有 最近被克隆和鉴定，包括胞浆磷脂酶 A2(CPLA2)和环氧合酶(COX-L和COX-2)。最近的证据 提示cPLA2和COX可能具有不同的本构关系和 荷尔蒙调节的异构体，以及表达的变化 这些酶由IL-I和肿瘤坏死因子在几个细胞系统中诱导，包括 欧共体。然而，被调控的精确异构体和基因的变化 细胞因子处理EC中的酶活性尚不清楚 明白了。在这笔赠款期间，我将进行有系统的 了解这些基因调控的分子方面的方法 人脐静脉内皮细胞中肿瘤坏死因子和白介素1的酶活性分析 信使核糖核酸转录速率、信使核糖核酸和蛋白质的稳定性和水平，以及 酶活性的变化。将利用的技术包括核能 径流、北方印迹、Western印迹、免疫沉淀和 生化酶分析。这些研究应该提供一个完整的 这一关键生物学途径的细胞因子调控的描述 欧共体。

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.