REG OF THE NADPH OXIDASE BY ANTI-INFLAMMATORY AGENTS

本计画的长期目标将是研究吞噬白血球产生超氧化物的NADPH氧化酶的调控。 由该酶系统产生的氧物种在宿主防御感染以及在炎症和再灌注损伤期间的组织损伤中起主要作用。 我们的工作将集中在NADPH氧化酶的细胞色素B组分上，特别是编码gp 91-phox的基因，gp 91-phox是吞噬细胞细胞色素B异二聚体的91千道尔顿糖蛋白。 该基因和密切相关基因的表达受发育调控，并且几乎对骨髓单核细胞具有谱系特异性。 糖皮质激素和非甾体抗炎药（NSAID）调节各种基因和细胞功能的表达。 拟定的研究将检验中心假设，即糖皮质激素和NSAID通过下调编码NADPH氧化酶系统组分的基因表达来抑制吞噬细胞NADH氧化酶活性。 我们将：1.通过影响吞噬细胞功能的细胞因子开发用于分化人单核细胞系THP-1的系统：我们将检测NADPH氧化酶活性、细胞色素B含量以及编码gp 91-phox和其他NADPH氧化酶组分的基因的表达。 2.研究糖皮质激素和非甾体类抗炎药（NSAID）对姜黄素分化的THP-1细胞NADPH氧化酶系统的影响：我们将检测糖皮质激素和NSAID对NADPH氧化酶活性、细胞色素B含量以及编码gp 91-phox和其他NADPH氧化酶组分的基因表达的影响。 3.研究细胞因子、糖皮质激素或NSAID对人外周血单核细胞/巨噬细胞中NADPH氧化酶系统调节的影响：我们将研究糖皮质激素或NSAID对细胞因子刺激下人外周血单核细胞/巨噬细胞中NADPH氧化酶活性、细胞色素B含量以及编码gp 91-phox和其他NADPH氧化酶组分的基因表达的影响。拟议的研究应推进我们的基本知识的转录控制机制在骨髓单核细胞谱系，以及确定新的机制，抗炎剂发挥其抗炎作用，在这种情况下，通过下调基因的表达编码吞噬NADPH氧化酶系统的组件。 更好地了解NADPH氧化酶系统的药理学调节将提供信息，以帮助调节其在炎症和氧化剂介导的组织损伤过程中的表达。

The long-term goal of this project will be to investigate the regulation of the superoxide-generating NADPH oxidase of phagocytic leukocytes. Oxygen species generated by this enzyme system play a major role in host defense against infection and as well as in tissue damage during inflammation and reperfusion injury. Our work will focus on the cytochrome b component of the NADPH oxidase, in particular the gene encoding gp91-phox, the 91 kilodalton glycoprotein of the phagocyte cytochrome b heterodimer. Expression of this and closely related genes is developmentally regulated and virtually lineage-specific to myelomonocytic cells. Glucocorticoids and non-steroidal anti-inflammatory drugs (NSAID) modulate the expression of various genes and cell functions. The proposed studies will test the central hypothesis that glucocorticoids and NSAID inhibit phagocytic NADH oxydase activity by down-regulating expression of genes encoding components of the NADPH oxidase system. We will: 1. Develop a system for the differentiation of the human monocytic cell line THP-1 by cytokines that influence phagocyte function: We will examine the NADPH oxidase activity, cytochrome b content, and expression of the genes encoding gp91-phox and the other NADPH oxidase components. 2. Study the effects of glucocorticoids and NSAID on the NADPH oxidase system of cytokine-differentiated THP-1 cells: We will examine the effect of glucocorticoids and NSAID on the NADPH oxidase activity, cytochrome b content, and expression of the genes encoding gp91-phox and the other NADPH oxidase components. 3. Investigate the effects of cytokines, glucocorticoids or NSAID on the regulation of the NADPH oxidase system in human peripheral blood monocytes/macrophages: We will investigate the effects of glucocorticoids or NSAID on the NADPH oxidase activity, cytochrome b content, and expression of the genes encoding gp91-phox and the other NADPH oxidase components in human peripheral blood monocytes/macrophages under cytokine stimulation. The proposed studies should advance our fundamental knowledge of transcriptional control mechanisms in the myelomonocytic lineage and as well determine new mechanisms by which anti-inflammatory agents exert their anti-inflammatory effects, in this case by down-regulating the expression of genes encoding components of the phagocytic NADPH oxidase system. Better understanding of the pharmacological regulation of the NADPH oxidase system will provide information to help modulate its expression during inflammation and oxidant-mediated tissue damage.