MicroRNA expression profile in the third- and fourth-stage larvae of Angiostrongylus cantonensis
MicroRNA expression profile in the third- and fourth-stage larvae of Angiostrongylus cantonensis
复制标题
广州管圆线虫三、四期幼虫中MicroRNA的表达谱
DOI：
10.1007/s00436-014-3836-6
复制
发表时间：
2014-05-01
影响因子：
2
通讯作者：
Wu, Zhong-Dao
中科院分区：
文献类型：
--
作者：
Li, Zhengyu;Chen, Xiaoguang;Wu, Zhong-Dao
广州管圆线虫病是由广州管圆线虫第三、四期幼虫侵入人体中枢神经系统引起的，其发病机制至今仍不清楚。microRNA是基因表达的重要调控因子，参与多种生物学过程。本研究的目的是确定和表征的miRNAs的第三（L3）和第四（L4）的幼虫。通过Solex deep sequencing。共检测到629个保守的miRNAs（L3和L4幼虫中分别有526和376个miRNAs）。cantonensis）和三个新的候选miRNA。通过生物信息学分析，鉴定了广州亚种（Cantonensis）。在A的L4中有163个miRNAs表达上调，54个miRNAs表达下调（倍数变化≥5.0）。cantonensis与A.广东话。有趣的是，Gene Ontology的“生物过程”分类揭示了26种显著差异表达的miRNAs与免疫系统相关，这意味着这些miRNAs可能通过调节参与免疫反应途径的基因参与管圆线虫病的发病机制。此外，还对26种保守的miRNA在A.广州人的身份得到了验证。real-time PCR和北方印迹分析结果表明，aca-miR-124和aca-miR-146 a-5 p在L3幼虫中低水平表达，而在L4幼虫中高水平表达。与aca-miR-124模拟物和抑制剂联合转染相比，aca-miR-124模拟物单独转染显著下调N9细胞中IL-6、IL-1β和TNF-α的mRNA表达（P <0. 05），提示A. cantonensis在抑制小胶质细胞活化中具有重要作用。总之，本研究提供了小RNA在A.并突出了L3和L4幼虫之间差异表达的保守miRNA。我们的数据表明，寄生虫的miRNA可能介导的重要作用，在A。cantonensis免疫逃避和aca-miR-146 a-5 p可作为诊断广州管圆线虫病的潜在生物标志物。
The pathogenesis of angiostrongyliasis, resulting from the third-stage and the fourth-stage Angiostrongylus cantonensis larvae invasion of the human central nervous system, remains elusive. MicroRNAs are important regulators of gene expression and involved in many biological processes. The aim of this study was to determine and characterize miRNAs of third (L3) and fourth (L4) larvae of A. cantonensis by Solex deep sequencing. A total of 629 conserved miRNAs (526 and 376 miRNAs in L3 and L4 larvae of A. cantonensis, respectively) and three novel candidate miRNA from L3 and L4 larva of A. cantonensis were identified with bioinformatic analysis. There were 163 miRNAs upregulated and 54 miRNAs downregulated (fold changes ≥5.0) in the L4 of A. cantonensis compared with that of L3 of A. cantonensis. Interestingly, Gene Ontology "biological process" classifications revealed that 26 miRNAs of significantly differential expression are associated with the immune system, which implies that these miRNAs might participate in the pathogenesis of angiostrongyliasis by regulating genes involved in immune response pathways. Furthermore, the differential expression patterns of 26 conserved miRNAs between L3 and L4 of A. cantonensis were verified. The results of real-time PCR and Northern blot showed that the aca-miR-124 and aca-miR-146a-5p have a low level expression in L3 larvae but high level expression in L4 larvae. Transfection of aca-miR-124 mimics alone significantly downregulated the mRNA expression of IL-6 and IL-1β and TNF-a in the N9 cells, compared to the combination transfection of aca-miR-124 mimics and inhibitor (P < 0.05), suggesting that miR-124 of A. cantonensis have an important role in the suppression of microglia activation. In conclusion, the study presents a general picture of the expression of small RNAs in L3 and L4 of A. cantonensis and highlights conserved miRNAs differentially expressed between L3 and L4 larvae. Our data revealed that miRNAs of parasite may mediate important roles in A. cantonensis immune evasion and aca-miR-146a-5p can serve as a potential biomarker to diagnose angiostrongyliasis.