CELLULAR REGULATION OF PROSTAGLANDIN SYNTHESIS

描述(改编自《调查者摘要》)：这是一场竞争 续签赠款，该基金几年来一直在审查 前列腺素内酯过氧化合成酶(PGHS)的调节。肾小球系膜 细胞(GMC)在几种肾小球疾病的发生中起关键作用 如肾小球硬化和增生性肾小球肾炎，它们是 以GMC损伤和增殖为特征。促炎药可以 诱导细胞合成二十烷类化合物，即氧化产物 花生四烯酸，可调节GMC基质的合成和增殖。 这个应用程序检查细胞内信号机制调节 催化细胞产生前列腺素的PGHS的表达和作用 花生四烯酸。PGHS特别令人感兴趣，成为 非甾体抗炎药。将使用体外和体内研究相结合的方法来测试 有丝分裂原激活蛋白调控PGHS-2表达的假说 蛋白激酶(MAPK)和PGHS-2的过度表达具有抗细胞凋亡的作用 通过差异基因表达来调节。 已勾勒出三个具体目标。特定目标1将评估 假设MAPK的激活代表了 对PGHS-2表达的不同刺激，这是负调控的 双特异性磷酸酶。在这里，对PGHS-2表达的影响 腺病毒介导的野生型和突变型ERK、JNK基因转移 和p38MAPKs，以及肾脏中的双特异性磷酸酶 细胞。特定目标2将检验PGHS-2表达导致 在几种细胞类型中发挥抗凋亡作用。这里的效果是 腺病毒介导的PGHS-2基因导入原代细胞诱导的细胞凋亡 将对参数进行研究。其抗细胞凋亡作用的机制将是 利用cDNA鉴定PGHS-2表达调控基因的研究 微阵列。在特定目标3中，PGHS-2的表达与体内的相关性 与MAPK级联反应的关系将使用两种大鼠模型进行评估 肾小球肾炎(GN)在这里，PGHS-2的表达与前列腺素的产生 将在大鼠肾小球中进行评估，并与MAPK激活和 明确疾病时间点的肾小球肾炎的组织病理学。

DESCRIPTION (Adapted from Investigator's Abstract): This is a competitive renewal of a grant, which has for several years examined the mechanisms of regulation of prostaglandin endoperoxide synthase (PGHS). Glomerular mesangial cells (GMC) play a key role in the genesis of several glomerular diseases such as glomerulosclerosis and proliferative glomerulonephritis, which are characterized by GMC injury and proliferation. Proinflammatory agents can induce cellular synthesis of eicosanoids, namely oxygenated products of arachidonic acid, that can regulate GMC matrix synthesis and proliferation. This application examines the intracellular signaling mechanisms regulating the expression and actions of PGHS, which catalyses prostaglandin production from arachidonic acid. PGHS is of particular interest, being the principal target of NSAIDS. A combination of in vitro and in vivo studies will be used to test the hypothesis that PGHS-2 expression is regulated by mitogen activated protein kinases (MAPKs), and that over expression of PGHS-2 has anti-apoptotic effects mediated through differential gene expression. Three Specific Aims have been outlined. Specific Aim 1 will evaluate the hypothesis that activation of MAPKs represents the convergence point of divergent stimuli of PGHS-2 expression, and this is negatively regulated by dual-specificity phosphatases. Here, the effects on PGHS-2 expression after adenovirus mediated transfer of genes encoding wild type and mutant ERK, JNK and p38 MAPKs, and dual-specificity phosphatases will be studied in kidney cells. Specific Aim 2 will examine the hypothesis that PGHS-2 expression leads to anti-apoptotic effects in several cell types. Here the effect of adenovirus-mediated transfer of PGHS-2 gene into primary cells on apoptotic parameters will be studied. The mechanism of the anti-apoptotic effect will be studied by identifying genes controlled by PGHS-2 expression using cDNA micro-arrays. In Specific Aim 3, the in vivo relevance of PGHS-2 expression and relation to the MAPK cascade will be evaluated using two rat models of glomerulonephritis (GN). Here, PGHS-2 expression and prostaglandin production will be assessed in rat glomeruli and correlated with MAPK activation and the histopathology of GN at defined time points of the disease.