目的 建立HBsAg刺激树突状细胞(DC)前后microRNA (miRNA)表达的差异谱,为研究miRNA在乙型肝炎病毒感染的免疫机制中的作用提供理论基础.方法 体外培养原代DC,分别用HBsAg、LPS、TNFα刺激24 h后提取细胞总RNA,利用基因芯片技术进行比较,筛选出HBsAg刺激后差异表达显著的miRNA,利用计算机软件技术进行靶基因初步预测.结果 HBsAg刺激前后miRNA差异表达2倍以上者共30个,其中16个上调,14个下调.三个不同刺激组间miRNA差异谱存在不同差异.筛选出的靶基因中包含DC信号通路的相关成分.结论 HBsAg刺激DC后的miRNA表达谱的变化存在特异性;靶基因的初步筛选结果进一步提示了miRNA在乙型肝炎病毒感染的免疫机制中可能起着重要作用.%Objective To establish the differential expression profiles for exploring new immune mechanism of hepatitis B virus infection.Methods DCs were separated from the bone marrow of healthy mouse and cultured in vitro.Then DCs were stimulated with HBsAg,LPS,TNF-α,respectively.Twentyfour hours later,the total RNA of cell was extracted.cDNA microarray was used to compare the differential expression of RNA.The significant differential expression of miRNA after the stimulation of HBsAg was chosen.Subsequently,target genes of the significant differential miRNA were forecasted by using computer software.Results There were 30 miRNAs whose significant differential expressions were beyond two times after the stimulation of HBsAg.Among them,16 miRNAs expressions were increased and 14 miRNAs expressions were decreased.There was significant difference in differential expression of miRNA among the 3 different stimulations.The selected target genes included relevant elements of signal pathway of DC.Conclusion The alteration of expression profiles of miRNA was specific after DC was stimulated by HBsAg.The selected target genes further demonstrated that miRNA could play important roles in the immune mechanism of HBV infection.
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