【摘要】 目的:初步探讨伴侣蛋白C1GALT1C1在IgA肾病(IgA nephropathy,IgAN)发病中的作用及从患者外周血单核细胞中筛选靶向C1GALT1C1基因的miRNA。方法:收集入住我院的IgAN和健康成人外周抗凝血,分离获取单核淋巴细胞,各分成两份。一份用于提取单核淋巴细胞细胞总蛋白行免疫印迹实验检测C1GALT1C1蛋白水平。慢病毒载体介导DAKIKI细胞发生C1GALT1C1基因沉默并检测细胞分泌的糖基化低下的IgA1水平。另一份IgAN和健康成人外周血单核淋巴细胞用于miRNA表达谱测序、生物信息学分析与靶向C1GALT1C1基因的miRNA筛选。结果:IgAN患者外周单核淋巴细胞C1GALT1C1蛋白平均水平下调约40%;慢病毒载体介导DAKIKI细胞发生C1GALT1C1基因沉默后细胞分泌的糖基化水平低下IgA1水平升高;miRNA表达谱发现有123个差异表达在2倍以上的miRNA,其中有25个表达上调,它们的预测靶基因都为C1GALT1C1。结论:伴侣蛋白C1GALT1C1的低表达可能对IgAN发病起了一定的作用;从IgAN患者外周血单核细胞中筛选到靶向C1GALT1C1基因的25个miRNA对于该病的诊断和预后可能有一定的指导意义。更多还原

【Abstract】 Objective:To preliminarily investigate the role of chaperone C1 GALT1 C1 in the pathogenesis of IgAN and to screen miRNAs targeting the C1 GALT1 C1 gene from peripheral blood monocytes of IgAN patients.Methods:Peripheral anticoagulant blood was collected from IgAN patients and healthy controls,and mononuclear lymphocytes were isolated and divided each into two parts. One was used to extract the total protein of mononuclear lymphocytes and conduct west-ern blot assay to detect the protein level of C1 GALT1 C1. After lentiviral vectors mediating C1 GALT1 C1 gene silencing in DAKIKI cells,underglycosylation IgA1 levels in cell secretion were detected by ELISA. Sequencing,bioinformatics analysis and screening of miRNAs targeting C1 GALT1 C1 gene in another peripheral blood mononuclear lymphocytes of IgAN patients and healthy controls were applied.Results:The average level of C1 GALT1 C1 protein in peripheral mono-cytes of IgAN patients was down-regulated by about 40%. Lentivirus vector mediated the increase of underglycosylated IgA1 in DAKIKI cells after C1 GALT1 C1 gene silencing. The miRNA expression profile found that 123 miRNAs differen-tially expressed more than two times,among which 25 were up-regulated and their predicted target genes were C1 GALT1 C1.Conclusion :The low expression of chaperone C1 GALT1 C1 may play a role in the pathogenesis of IgAN.Twenty-five miRNAs targeting the C1 GALT1 C1 gene were screened from peripheral blood mononuclear cells of IgAN pa-tients,which may have some guiding significance for the diagnosis and prognosis of this disease.更多还原