CORNEAL ARACHIDONIC ACID METABOLISM

这笔赠款的目的是确定：(1)花生四烯酸如何 (Aa)正常大鼠角膜上皮细胞的代谢受到调节；(2)什么 角膜灼伤后花生四烯酸代谢发生改变； (3)非甾体抗炎药能否预防角膜 通过抑制AA代谢而引起炎症和血管形成。监管 AA代谢的研究将在分离的完整上皮中进行 中性蛋白水解酶治疗角膜。细胞磷脂将是 标记有[~3H]AA和各种物质将进行测试，以确定 如果它们激活磷脂酶A2，从而释放游离AA。 磷脂酶A2激活是调节AA代谢的关键步骤 并且有可能受到药物抑制。另一个项目 将用硝酸钾/硝酸银烧灼大鼠角膜 调查在甲型H1N1流感流入前AA代谢的变化 炎性细胞。外源[~3H]AA的代谢将被量化 此外，我们还将开发一种用于测量 L-3-磷脂酰胆碱，1-硬脂酰基-2-[~3H]花生四烯基代谢。这些 两种互补的方法将使我们能够检测到受伤引起的变化 磷脂酶A2激活与脂氧合酶/环氧合酶途径 活动。最后一项研究将检验脂氧合酶的影响 抑制剂，环氧合酶抑制剂，和双重 环氧合酶-脂氧合酶抑制剂对角膜炎症和角膜炎症的影响 大鼠角膜灼伤后的血管形成。机管局的变更 炎症和血管生成反应过程中的新陈代谢 仅使用生理盐水(药物载体)处理的动物进行研究。 这些研究将确定AA代谢物是否是重要的介体 角膜炎症和血管形成，并将阐明潜在的 非甾体抗炎药在治疗角膜中的作用 受伤。

The objectives of this grant are to determine: (1) how arachidonic acid (AA) metabolism is regulated in normal rat corneal epithelium; (2) what changes occur in arachidonate metabolism following corneal cauterization; and (3) whether nonsteroidal anti-inflammatory drugs can prevent corneal inflammation and vascularization by inhibiting AA metabolism. Regulation of AA metabolism will be studied in intact epithelium isolated by neutral-protease treatment of corneas. Cellular phospholipids will be labeled with [3H]AA and a variety of substances will be tested to determine if they activate phospholipase A2 with resultant release of free AA. Phospholipase A2 activation is a key step in regulation of AA metabolism and is potentially amenable to pharmacological inhibition. Another project will use potassium/silver nitrate cauterization of rat corneas to investigate changes in AA metabolism occurring prior to the influx of inflammatory cells. Metabolism of exogenous [3H]AA will be quantitated and, in addition, we will develop an assay for measuring L-3-phosphatidylcholine, 1-stearoyl-2-[3H]arachidonyl metabolism. These two complimentary approaches will allow us to detect injury-induced changes in phospholipase A2 activation and lipoxygenase/cyclooxygenase pathway activity. A final study will examine the influence of lipoxygenase inhibitors, cyclooxygenase inhibitors, and a dual cyclooxygenase-lipoxygenase inhibitor on corneal inflammation and vascularization following cauterization of rat corneas. Changes in AA metabolism during the inflammatory and vascularization responses will also be studied using animals treated with normal saline (drug vehicle) only. These studies will determine whether AA metabolites are important mediators of corneal inflammation and vascularization and will clarify the potential usefulness of nonsteroidal antiinflammatory drugs for treating corneal injuries.