STUDIES OF PHOSPHOLIPASE A2 GENE EXPRESSION

胞浆磷脂酶A2（cPLA 2）是一种花生四烯酸特异性磷脂酶， 磷脂酶在类花生酸的细胞生产中起重要作用。细胞因子 在气道上皮细胞中研究cPLA 2活性的活化。它 发现T淋巴细胞因子干扰素γ引起的 从气道上皮细胞释放花生四烯酸。Western blot 分析显示，在用抗肿瘤药物治疗的上皮细胞中，cPLA 2增加。 干扰素-γ治疗4-24小时。稳态mRNA水平的测定 在相同的时间范围内显示cPLA 2 mRNA水平增加， 转录分析表明，在转录增加， cPLA 2 mRNA对干扰素刺激的响应率。同样，TNF 气道上皮细胞系的处理导致 花生四烯酸释放Western印迹显示cPLA 2蛋白增加。 cPLA 2 mRNA的核糖核酸酶保护试验显示， 4-24小时的状态消息。转录检测显示 TNF处理后cPLA 2 mRNA的转录速率。 白细胞介素-6能够激活某些炎症中的PLA 2活性 细胞另一种利用gp 130作为其受体的一部分的细胞因子 白血病抑制因子（LIF）气道上皮的蛋白质印迹 细胞株表达LIF受体蛋白。LIF治疗这些细胞 导致花生四烯酸释放增加。蛋白质印迹分析 细胞cPLA 2蛋白的表达揭示了蛋白质的增加， LIF治疗。cPLA 2稳态mRNA的核糖核酸酶保护测定 水平显示cPLA 2信息在治疗4-24小时内增加 关于LIF正在进行的研究将评估cPLA 2的细胞因子控制 活性通过基因表达和翻译后机制。一个 对cPLA 2细胞因子激活机制的理解可能有助于 新的呼吸道疾病的治疗。其中一份手稿 已出版并编写了两份手稿。

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-gamma for 4-24 hours. Assays for steady state mRNA levels demonstrated an increase in cPLA2 mRNA levels over the same time frame and transcriptional analysis demonstrated an increase in the transcription rate of cPLA2 mRNA in response to Interferon stimulation. Similarly, 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 4-24 hours. Transcriptional assay reveals an increase 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 which utilizes gpl30 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 an increase in 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 reveal an increase in cPLA2 message over 4-24 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 diseases of the airways. One manuscript has been published and two manuscripts have been prepared.