乙型肝炎表面抗原对RAW264.7细胞株Toll样受体表达模式的影响  

The effects of HBsAg on the expression of Toll like receptors in Raw264.7 cells

作　　者：杜玲[1] 田怡[1] 刘建生[1] 

机构地区：[1]上海瑞金医院集团闵行区中心医院消化科,上海201508

出　　处：《中国病毒病杂志》2013年第1期51-54,共4页Chinese Journal of Viral Diseases

基　　金：上海市卫生局青年基金资助项目(2009021);上海市闵行区科委项目(2011MHZ11)

摘　　要：目的探讨乙肝表面抗原(HBsAg)对Toll样受体(TLR)表达的影响效应以及TLR激动剂对这一效应的干预作用。方法以HBsAg与RAW264.7细胞共孵育,在有或无TLR激动剂[PGN(TLR2配体)、PolyIC(TLR3配体)、LPS(TLR4配体)、R848(TLR7/8配体)、CpG(TLR9配体)]刺激下,以RT-real time PCR检测Toll样受体TLR2、3、4、7、9的基因表达情况;同时,用流式细胞技术检测HBsAg和LPS单独或共刺激的Raw264.7细胞TLR表达情况。结果定量PCR检测和流式细胞分析均表明,HBsAg导致TLR3表达显著下调(P<0.05),其中TLR3基因表达下调80%,受体表达也显著下调(P<0.05);对其他4个TLR表达也有一定下调作用,但差异无统计学意义。加入Toll样受体激动剂后,TLR3和TLR9基因表达显著上调(P<0.01)。TLR4基因表达也有部分上调作用。流式细胞技术检测表明,HBsAg+LPS显著上调TLR3和TLR9的表达水平(P<0.01)。结论 HBsAg在细胞水平能显著抑制Toll样受体的表达,且Toll样受体激动剂能改善HBsAg的抑制效应。Objective To study the effects of HBsAg with or without Toll-like receptor（TLR）agonists on the expression of TLR in Raw264.7 cell line.Methods After incubation of RAW264.7 cells and HBsAg with or without TLR agonists,the expression of TLR 2,3,4,7,9 was detected using RT-Real-time PCR.Meanwhile,HBsAg and/or LPS treated RAW264.7 cells were collected for the fluorescence correlation spectroscopy（FCS）analysis of TLR.Results In the presence of HBsAg,gene expression of TLR3 was significantly downregulated（P0.05）,and varied down-regulation of the other four TLRs were also observed（P0.05）.All TLR agonists significantly increased the gene expression of both TLR3 and TLR9 on HBsAg treated cells（P0.01）,while TLR agonists showed various effects on the upregulation of TLR4 gene expression.FCS analysis demonstrated similar results of TLRs expression in Raw264.7 cells.Conclusions HBsAg significantly inhibits the expression of TLRs,and TLR agonists could reverse this effects.

关 键 词：乙型肝炎病毒 乙型肝炎表面抗原 REAL-TIME PCR TOLL样受体 

分 类 号：R512.62[医药卫生—内科学]