STUDIES OF PHOSPHOLIPASE A2 GENE EXPRESSION

胞浆磷脂酶A2(CPLA2)是花生四烯酸专一性的 磷脂酶在类二十碳化合物的细胞生产中起重要作用。 细胞因子激活对呼吸道cPLA2活性的影响 上皮细胞。研究发现，T淋巴细胞细胞因子 干扰素-γ引起呼吸道花生四烯酸释放 上皮细胞。Western印迹分析显示， 干扰素-g作用于上皮细胞4~24小时的cPLA2 几个小时。对稳态信使核糖核酸水平的检测显示增加 同时检测cPLA2基因的表达水平。转录分析 显示cPLA2基因转录速率增加。 对干扰素刺激的反应。同样，肿瘤坏死因子 (肿瘤坏死因子)处理呼吸道上皮细胞系导致 激活花生四烯酸释放。西方印迹揭示 CPLA2蛋白增加。CPLA2的核糖核酸酶保护试验 在4-24小时内，信使核糖核酸表达增加。 转录分析显示转录速度增加 CPLA2基因对肿瘤坏死因子处理的反应。 白介素6能够激活某些细胞的PLA2活性 炎性细胞。另一种使用gp130的细胞因子 其受体为白血病抑制因子(LIF)。An的蛋白质印迹 呼吸道上皮细胞株表达LIF受体蛋白。寿命 对这些细胞的处理会导致花生四烯酸增加 放手。细胞cPLA2蛋白的Western印迹分析显示 对激光诱导因子治疗的反应增加了蛋白质。核糖核酸酶 对cPLA2稳态mRNA水平的保护实验显示 在LIF治疗4至24小时后，cPLA2消息增加。 正在进行的研究将评估细胞因子对cPLA2活性的控制 通过基因表达和翻译后机制。一个 了解细胞因子激活cPLA2的机制可能 允许对呼吸道疾病进行新的治疗方法。二 手稿已经出版，一份手稿已经出版 准备好了。

Cytosolic phospholipase A2 (cPLA2) is an arachidonate-specific phospholipase important in cellular production of eicosanoids. Cytokine activation of cPLA2 activity was studied in airway epithelial cells. It was found that the T lymphocyte cytokine Interferon-gamma caused arachidonic acid release from airway epithelial cells. Western blot analysis revealed an increase in cPLA2 in epithelial cells treated with Interferon-g for 4 to 24 hours. Assays for steady-state mRNA levels demonstrated increased cPLA2 mRNA levels over the same time. Transcriptional analysis demonstrated an increase in the transcription rate of cPLA2 mRNA in response to Interferon stimulation. Similarly, tumor necrosis factor (TNF) treatment of an airway epithelial cell line results in activation of arachidonic acid release. Western blots reveal increases in cPLA2 protein. Ribonuclease protection assay for cPLA2 mRNA reveals increases in steady-state message for 4 to 24 hours. Transcriptional assay reveals an increased rate of transcription of cPLA2 mRNA in response to TNF treatment. Interleukin-6 is capable of activating PLA2 activity in some inflammatory cells. Another cytokine that uses gp130 as a portion of its receptor is leukemia inhibitory factor (LIF). Western blot of an airway epithelial cell line revealed LIF receptor protein. LIF treatment of these cells results in increased arachidonic acid release. Western blot analysis of cellular cPLA2 protein reveals an increase in protein in response to LIF treatment. Ribonuclease protection assay for cPLA2 steady-state mRNA levels reveals an increase in cPLA2 message over 4 to 24 hours of treatment with LIF. Ongoing studies will assess the cytokine control of cPLA2 activity by gene expression and by post-translational mechanisms. An understanding of the mechanisms of cytokine activation of cPLA2 may allow for new therapeutic treatment of diseases of the airways. Two manuscripts have been published, and one manuscript has been prepared.