摘要
[目的]研究转染小凹蛋白-1(Caveolin-1,CAV-1)基因对脂多糖(LPS)诱导的人支气管上皮(HBE)细胞Toll样受体4(TLR-4)表达的影响,以及大黄素对CAV-1和TLR-4表达的干预效应。[方法]本实验分为2部分:第1部分,对LPS诱导的HBE细胞给予不同剂量的大黄素干预,并设立正常对照、阳性对照(阿托伐他汀)和阴性对照(溶媒DMSO),用Western blot检测各组细胞CAV-1的表达水平;第2部分,通过构建CAV1-EGFP重组质粒,瞬时转染HBE细胞,于转染48h后给予LPS刺激,并给予大黄素干预转染后LPS诱导的HBE,Western blot检测各组细胞TLR-4的表达。[结果]中、高剂量的大黄素均能抑制LPS诱导的HBE细胞CAV-1的表达;与各对照组相比,CAV-1过表达能上调LPS诱导的HBE细胞TLR-4的表达,大黄素能抑制该效应。[结论]转染CAV-1基因能上调LPS诱导的HBE细胞TLR-4的表达,大黄素能抑制LPS诱导的HBE细胞CAV-1与TLR-4的表达。
[Objective]To investigate the effect of Caveolin-1（CAV-1） gene transfection on the expression of TLR-4 in LPS-induced HBE cells and the intervention of Emodin on the expression of CAV-1 and TLR-4.[Methods]The experiment was divided into two parts.In the first part of the experiment,the protein of LPS-induced HBE cells from different intervention groups was extracted to measure the expression of CAV-1 by western blot.In the second part of the experiment,the CAV1-EGFP recombinant plasmid was transiently transfected into the HBE cells.Thereafter,LPS was administered into the cell culture medium at 48h post-transfection,and Emodin was given into the LPS-induced HBE cells after transfection.Then the expression of TLR-4 was assayed by western blot.[Results]Both the middle-dose Emodin and the high-dose Emodin suppressed the expression of CAV-1 in LPS-induced HBE cells.Compared with that of control groups,CAV1-EGFP overexpression up-regulated the expression of TLR-4 in LPS-induced HBE cells and Emodin inhibitted this effect.[Conclusion]CAV-1 gene transfection could up-regulated the expression of TLR-4 in LPS-induced HBE cells.Emodin could inhibit the expression of CAV-1 and TLR-4 in LPS-induced HBE cells.
出处
《中国中西医结合消化杂志》
CAS
2012年第9期405-409,共5页
Chinese Journal of Integrated Traditional and Western Medicine on Digestion
基金
国家自然科学基金资助项目(No:30873225)
湖北省卫生厅科研基金资助课题(No:2010z-y20)
