發(fā)布時間：2018-02-27 04:34

  本文關鍵詞： emodin 核因子-κB 抑制蛋白-κB I-κB激酶 RNA干擾 脂多糖 誘生性一氧化氮合酶 腫瘤壞死因子-α 白介素-10 巨噬細胞 急性胰腺炎 急性肺損傷 一氧化氮 中西醫(yī)結合　出處：《大連醫(yī)科大學》2006年博士論文　論文類型：學位論文

【摘要】：目的： 探討核因子-κB炎癥信號通路在全身炎癥反應時肺泡巨噬細胞過度活化中的作用，觀察NF-κBp65／p50的活性和核易位情況、蛋白激酶IKK的活性變化、炎性細胞因子的表達水平，并深入探究蛋白激酶IKKαγ對上述炎癥信號通路中多個環(huán)節(jié)的協(xié)同調節(jié)效應及Emodin治療作用的分子生物學機制；同時，進一步揭示一氧化氮(NO)在急性重癥胰腺炎急性肺損傷大鼠發(fā)病中的雙重功能及該作用與肺泡巨噬細胞NF-κB活性變化的密切聯(lián)系，為臨床治療急性肺損傷等多種全身炎癥反應性疾病、尋求更為合適的藥物作用靶點提供新的實驗佐證。 方法： 實驗一：以小鼠RAW264.7巨噬細胞系為研究對象，應用逆轉錄PCR、蛋白雜交印跡、免疫細胞化學染色、免疫熒光染色等方法，觀察emodin對由LPS誘導產生的炎癥反應的調節(jié)作用。共分為7組，每組5個培養(yǎng)皿：Group 1：RAW264.7巨噬細胞正常培養(yǎng)組；Group 2、3、4：給予LPS(101μg／ml)刺激，分三個時間點30min、2hr、5hr；Group 5、6：分為三個時間點，在給予LPS(10μg／ml)刺激的基礎上，，同時加入emodin(20gμg／ml)進行干預，于不同作用時間點收集的樣本。 實驗二：由NF-κB介導的過度炎癥反應在許多炎性疾病的發(fā)生發(fā)展過程中發(fā)揮著舉足輕重的作用，是全身炎癥反應時復雜病理變化的中心環(huán)節(jié)，同時也是多種因素、多層面、多環(huán)節(jié)協(xié)同作用的結果。為進一步闡明上述協(xié)同作用的分子生物學機制，應用RNA干擾技術將IκB蛋白激酶α及γ基因沉默，然后觀察RAW264.7小鼠巨噬細胞經LPS刺激后，NF-κB的活化以及多種NF-κB依賴性的
[Abstract]:Objective:. To investigate the role of nuclear factor- 魏 B inflammatory signaling pathway in the excessive activation of alveolar macrophages during systemic inflammatory reaction, to observe the activity and nuclear translocation of NF- 魏 Bp65/p50, the changes of protein kinase IKK activity and the expression of inflammatory cytokines. The co-regulation effect of protein kinase IKK 偽 緯 on many parts of the above inflammatory signaling pathway and the molecular biological mechanism of Emodin therapy were also investigated. To further explore the dual function of nitric oxide (no) in the pathogenesis of acute lung injury in rats with severe acute pancreatitis and its close relationship with the change of NF- 魏 B activity in alveolar macrophages. To provide new experimental evidence for clinical treatment of various systemic inflammatory and reactive diseases such as acute lung injury and seeking more suitable drug targets. Methods:. Experiment 1: mouse RAW264.7 macrophage cell lines were studied by reverse transcription-PCR, Western blot, immunocytochemical staining, immunofluorescence staining and so on. To observe the effect of emodin on the inflammatory response induced by LPS, we divided into 7 groups, 5: 1: RAW264.7 macrophage normal culture group (n = 5: 1: RAW264.7) were stimulated by LPS(101 渭 g / ml, and they were divided into three groups at 30 min. On the basis of the stimulation of LPS(10 渭 g / ml, emodin(20g 渭 g / ml was added at the same time, and the samples were collected at different time points. Experiment two: the excessive inflammatory reaction mediated by NF- 魏 B plays an important role in the occurrence and development of many inflammatory diseases. It is the central link of complex pathological changes in systemic inflammatory reaction, and it is also a variety of factors and layers. In order to further elucidate the molecular mechanism of the synergistic effect, I 魏 B protein kinase 偽 and 緯 genes were silenced by RNA interference technique. Then, the activation of NF- 魏 B and various NF- 魏 B-dependent macrophages stimulated by LPS in RAW264.7 mice were observed.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2006
【分類號】：R363;R285

【參考文獻】

相關期刊論文 前2條

1 楊毅;吸入一氧化氮對急性肺損傷的治療作用—歐洲多中心研究報告[J];國外醫(yī)學.呼吸系統(tǒng)分冊;2000年03期

2 姜勇;內毒素激活內皮細胞的信號機制的研究進展[J];中華醫(yī)學雜志;1999年01期

本文編號：1541163