| 下调PALM3表达抑制脂多糖诱导的巨噬细胞炎性反应 |
| 投稿时间：2018-02-08 修订日期：2018-03-03 点此下载全文 |
| 引用本文：陈旭昕,宋立成,孟激光,韩志海.下调PALM3表达抑制脂多糖诱导的巨噬细胞炎性反应[J].医学研究杂志,2018,47(12):22-26 |
| DOI：
10.11969/j.issn.1673-548X.2018.12.007 |
| 摘要点击次数: 2028 |
| 全文下载次数: 1822 |
|
| 基金项目:国家自然科学基金资助项目（81300050）；北京市自然科学基金资助项目（7182163）；中国人民解放军海军总医院创新培养基金资助项目（CXPY201417） |
|
| 中文摘要:目的 探讨抑制paralemmin-3（PALM3）表达对脂多糖（LPS）诱导的大鼠肺泡巨噬细胞系（NR8383）炎性反应的影响。方法 将NR8383细胞并接种至6孔板中，待融合度达70%，加入0.5μg/ml LPS刺激12h，用免疫荧光检测细胞PALM3表达及亚定位；用0.5μg/ml LPS刺激NR8383细胞并在0、3、6、12、24和48h收集细胞提取总RNA，用RT-PCR法检测PALM3 mRNA；同前接种细胞至24孔板，待融合度达70%，将PALM3小干扰核糖核苷酸（siRNA）转染NR8383细胞（PALM3 siRNA转染组），同时设立正常NR8383细胞组和对照转染组（转染control siRNA），转染48h后，提细胞总蛋白用Western blot法检测PALM3蛋白水平；3组细胞给予LPS刺激12h，收集培养上清和核蛋白，用ELISA法检测培养上清中IL-8、IL-6的水平及核转录因子（NF-κB）转录活性。结果 PALM3分子在NR8383细胞中有表达，LPS可诱导PALM3表达；PALM3 siRNA转染后成功下调PALM3蛋白表达，予LPS刺激后，与正常细胞及对照转染组比较，PALM3 siRNA转染组培养上清中白介素-8（IL-8）和IL-6含量减少，PALM3 siRNA转染组细胞中NF-κB转录活性亦被抑制。结论 NR8383细胞中有PALM3表达，LPS可诱导NR8383细胞中PALM3的蛋白表达，下调PALM3表达可抑制LPS诱导的炎性反应。 |
| 中文关键词:Paralemmin-3 脂多糖 巨噬细胞 炎症 |
| |
| Downregulation of Paralemmin-3 Expression Inhibits the Inflammatory Reaction Induced by LPS in Alveolar Macrophages |
|
|
| Abstract:Objective To investigate the effect of downregulation of paralemmin-3 (PALM3) expression on the LPS-induced inflammatory reaction in rat alveolar macrophages (NR8383 cells). Methods NR8383 cells were conventionally cultured and seeded in 6-well plates. After the cell growth reached 70% confluence, NR8383 cells were treated with 0.5μg/ml for 12h. The expression and localization of PALM3 in NR8383 cells was determined by confocal immunofluorescence. And, the mRNA level of PALM3 were detected by RT-PCR at 0, 3, 6, 12, 24 and 48h post LPS-stimulation. Another NR8383 cells were seeded in 24-well plates. When the cell growth reached 70% confluence, the cells were transfected by PALM3 siRNA. Meanwhile, an empty vector was used as a negative control group and non-transfection NR8383 cells was used as a normal control group. At 48h after transfection, the protein expression of PALM3 in NR8383 cells was determined by Western blot analysis. LPS was administered into the cell culture medium at 48h post-transfection. The culture supernatant and nucleoprotein were collected at 12h after LPS-stimulation. The concentrations of interleukin-8 (IL-8) and IL-6 and the activity of nuclear transcrition factor-κB (NF-κB) were measured by ELISA. Results PALM3 was expressed in NR8383 cells and LPS could upregulate the PALM3 expression in a time-dependent manner. PALM3 siRNA could suppress the expression of PALM3 in NR8383 cells effectively. Downregulation of PALM3 expression could significantly inhibit the production of IL-8 and IL-6 and the activation of NF-κB induced by LPS in NR8383 cells. Conclusion PALM3 are expressed in rat alveolar macrophages (NR8383 cells) and LPS can cause PALM3-induced expression. Downregulation of PALM3 expression can ameliorate the inflammatory reactions to LPS in NR8383 cells. |
| keywords:Paralemmin-3 Lipopolysaccharide Alveolar macrophages Inflammation |
| 查看全文 查看/发表评论 下载PDF阅读器 |
