【作者】 钱琤；

【导师】 仲人前； 邓安梅； 韩焕兴； 谭龙益；

【作者基本信息】 第二军医大学 ， 临床检验诊断学， 2008， 博士

【摘要】 本研究旨在探讨原发性胆汁性肝硬化中T细胞microRNA(miRNA)表达谱及其调控机制。首先采用芯片和定量PCR技术分析并验证原发性胆汁性肝硬化中T细胞miRNA的表达谱;然后培养自身抗原PDC-E2特异性T细胞,研究特定miRNA对其自身反应性的调控作用;接着建立原发性胆汁性肝硬化小鼠模型,研究特定miRNA对小鼠PBC样病变的调控作用;最后通过生物信息学预测和生物学实验验证,探讨特定miRNA可能的靶基因及其作用位点。结果表明,与正常对照组相比,PBC患者T细胞中有23个miRNA低表达,仅有2个miRNA高表达;miR-1et-7b、miR-17-5p、miR-20a和miR-346表达可抑制自身抗原特异性T细胞增殖和IFN-γ、TNF-α的分泌,而miR-129和miR-451可促进细胞增殖和IFN-γ、TNF-α的分泌;在小鼠中输入Ad-miR-17-5p或Ad-miR-346可延缓由poly I:C诱发产生的PBC样病变;miR-17-5p、miR-346的靶基因分别为TNFRSF21、BCL-6,作用位点分别为TNFRSF21 3’UTR区335-342位和BCL-63’UTR区576-583及893-900位。更多还原

【Abstract】 To explore the RNA mechanism of PBC,we focus on the microRNA(miRNA) profile in the peripheral blood T cells of PBC and study on the action mechanism of miRNA in PBC.We first explored changes in the miRNA expression profile in the peripheral blood T cells of PBC patients by microarray and real-time PCR,then investigated the regulatory effects of some miRNA abnormally expressed in PBC on the proliferation of PDC-E2-specific autoreactive T cells.Subsequently,the effects of miRNA on the pathology of PBC disease were studied in animal models.Finally,miRNA target genes were predicted by bioinformatics and the effect of miRNA on the function of target genes was determined.The results showed that Twenty-three miRNA in the array were found to be expressed in low levels and two miRNA had high levels of expression in the peripheral blood T cells of patients with PBC.The proliferation and secretion of IFN-γ,TNF-αof autoantigen-specific T cells were inhibited by miR-let-7b,miR-17-5p,miR-20a,miR-346 and promoted by miR-129,miR-451.Injecting poly I:C into syngenic wild-type mice produced PBC-like pathological changes,which were alleviated following injection of Ad-miR-17-5p or Ad-miR-346.The target gene of miR-17-5p,miR-346 is TNFRSF21, BCL-6.The interaction sites are TNFRSF21 3’ UTR 335-342,BCL-6 3’ UTR 576-583 and 893-900.更多还原