下调paralemmin-3表达对脂多糖诱导的肺泡上皮细胞核转录因子-κB活性的影响

发布时间：2018-03-28 06:21

本文选题：急性肺损伤　切入点：内毒素　出处：《广东医学》2014年19期

【摘要】：目的研究下调paralemmin-3(PALM3)表达对脂多糖(LPS)诱导的肺泡上皮A549细胞核转录因子-κB(NF-κB)活性的影响。方法将针对PALM3的小干扰核糖核苷酸(siRNA)转染A549细胞后(PALM3siRNA转染组),用RT-PCR法检测PALM3 mRNA表达水平,同时设立control siRNA转染组及正常细胞组作为对照。转染48 h后,对3组细胞同时给予LPS刺激12 h后,收集细胞核蛋白和培养上清液,用ELISA的方法检测各组细胞培养上清液中的TNF-α、IL-6和IL-1β水平,用凝胶电泳迁移率实验的方法检测各组细胞NF-κB活性。结果特异性siRNA转染后成功下调PALM3在A549细胞中的表达;给予LPS刺激后,与正常细胞组及control siRNA转染组相比,PALM3 siRNA转染组细胞释放炎性因子TNF-α、IL-6和IL-1β减少,同时PALM3 siRNA转染组细胞中NF-κB活性受抑。结论下调PALM3的表达可减轻A549细胞对LPS诱导的炎症反应,调控PALM3的表达有望成为治疗急性肺损伤/急性呼吸窘迫综合征等炎症失控性疾病的新靶点。
[Abstract]:Objective to study the effect of down-regulation of paralemmin-3pALM3) expression on the activity of nuclear transcription factor- 魏 B-魏-魏 B in alveolar epithelium A549 induced by lipopolysaccharide (LPS). Methods small interfering ribonucleotide siRNAs targeting PALM3 were transfected into A549 cells and then the expression of PALM3 mRNA was detected by RT-PCR. At the same time, control siRNA transfection group and normal cell group were used as control. After 48 h of transfection, three groups of cells were stimulated with LPS for 12 h, then the nuclear protein and culture supernatant were collected. The levels of TNF- 伪 IL-6 and IL-1 尾 in the supernatant of each group were detected by ELISA method, and the activity of NF- 魏 B was detected by gel electrophoresis mobility assay. Results the expression of PALM3 in A549 cells was successfully down-regulated by specific siRNA transfection, and then stimulated by LPS. Compared with normal cell group and control siRNA transfection group, the release of inflammatory cytokines TNF- 伪, IL-6 and IL-1 尾 was decreased in PALM3 siRNA transfection group, while NF- 魏 B activity was inhibited in PALM3 siRNA transfection group. Conclusion down-regulation of PALM3 expression can reduce the inflammatory response induced by LPS in A549 cells. Regulation of PALM3 expression is expected to be a new target for the treatment of acute lung injury / acute respiratory distress syndrome (ARDS).
【作者单位】：中国人民解放军海军总医院呼吸内科;第三军医大学新桥医院呼吸内科;
【基金】：国家自然科学基金资助项目(编号:81300050)
【分类号】：R563.8

【参考文献】