论文摘要
目的： 探讨核因子-κB炎症信号通路在全身炎症反应时肺泡巨噬细胞过度活化中的作用，观察NF-κBp65／p50的活性和核易位情况、蛋白激酶IKK的活性变化、炎性细胞因子的表达水平，并深入探究蛋白激酶IKKα&γ对上述炎症信号通路中多个环节的协同调节效应及Emodin治疗作用的分子生物学机制；同时，进一步揭示一氧化氮（NO）在急性重症胰腺炎急性肺损伤大鼠发病中的双重功能及该作用与肺泡巨噬细胞NF-κB活性变化的密切联系，为临床治疗急性肺损伤等多种全身炎症反应性疾病、寻求更为合适的药物作用靶点提供新的实验佐证。 方法： 实验一：以小鼠RAW264.7巨噬细胞系为研究对象，应用逆转录PCR、蛋白杂交印迹、免疫细胞化学染色、免疫荧光染色等方法，观察emodin对由LPS诱导产生的炎症反应的调节作用。共分为7组，每组5个培养皿：Group 1：RAW264.7巨噬细胞正常培养组；Group 2、3、4：给予LPS（101μg／ml）刺激，分三个时间点30min、2hr、5hr；Group 5、6：分为三个时间点，在给予LPS（10μg／ml）刺激的基础上，同时加入emodin（20gμg／ml）进行干预，于不同作用时间点收集的样本。 实验二：由NF-κB介导的过度炎症反应在许多炎性疾病的发生发展过程中发挥着举足轻重的作用，是全身炎症反应时复杂病理变化的中心环节，同时也是多种因素、多层面、多环节协同作用的结果。为进一步阐明上述协同作用的分子生物学机制，应用RNA干扰技术将IκB蛋白激酶α及γ基因沉默，然后观察RAW264.7小鼠巨噬细胞经LPS刺激后，NF-κB的活化以及多种NF-κB依赖性的
论文目录
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