目的:研究大鼠坐骨神经缺损后背根神经节中microRNA(miRNA)的表达变化。方法:采用miRNA芯片检测神经缺损0 d(对照组)和7 d(实验组)背根神经节组织中miRNA的表达情况,应用Real time Taqman PCR对芯片结果进行验证。结果:实验组与对照组比较发生上调1.4倍和下调>1%以上的miRNA分别有13个和5个,Real time TaqmanPCR检测的4个miRNA的表达变化与芯片一致(上调:miR-21,miR-221;下调:miR-500,miR-551b)。结论:大鼠坐骨神经缺损1周后背根神经节组织中miRNA的表达谱发生改变。
Wallerian degeneration (WD) remains an important research topic. Many genes are differentially expressed during the process of WD, but the precise mechanisms responsible for these differentiations are not completely understood. In this study, we used microarrays to analyze the expression changes of the distal nerve stump at 0, 1, 4, 7, 14, 21 and 28 days after sciatic nerve injury in rats. The data revealed 6 076 differentially-expressed genes, with 23 types of expression, specifically enriched in genes associated with nerve development and axonogenesis, cytokine biosynthesis, cell differentiation, cytokine/chemokine production, neuron differentiation, cytokinesis, phosphorylation and axon regeneration. Kyoto Encyclopedia of Genes and Genomes pathway analysis gave findings related mainly to the MAPK signaling pathway, the Jak-STAT signaling pathway, the cell cycle, cytokine-cytokine receptor interaction, the p53 signaling pathway and the Wnt signaling pathway. Some key factors were NGF, MAG, CNTF, CTNNA2, p53, JAK2, PLCB1, STAT3, BDNF, PRKC, collagen II, FGF, THBS4, TNC and c-Src, which were further validated by real-time quantitative PCR, Western blot, and immunohistochemistry. Our findings contribute to a better understanding of the functional analysis of differentially-expressed genes in WD and may shed light on the molecular mechanisms of nerve degeneration and regeneration.
目的:研究大鼠坐骨神经缺损后损伤近端神经组织中microRNA(miRNA)的表达变化。方法:采用miRNA芯片检测神经缺损0h(对照组),3h和6h(实验组)损伤近端神经组织miRNA的表达情况,应用real time Taqman PCR对芯片结果进行验证。结果:3h实验组与对照组比较发生上调和下调1.5倍以上的miRNA分别有4个和3个,6h实验组与对照组比较发生上调和下调1.5倍以上的miRNA分别有11个和6个,real time TaqmanPCR检测miR-132和miR-223的表达变化与芯片检测结果一致。结论:大鼠坐骨神经缺损3h和6h后损伤近端神经组织中miRNA的表达谱发生改变。
The peripheral nervous system is able to regenerate after injury, and regeneration is associated with the expression of many genes and proteins. MicroRNAs are evolutionarily conserved, small, non-coding RNA molecules that regulate gene expression at the level of translation. In this paper, we focus on the identification and functional annotation of novel microRNAs in the proximal sciatic nerve after rat sciatic nerve transection. Using Solexa sequencing, computational analysis, and quantitative reverse transcription PCR verification, we identified 98 novel microRNAs expressed on days 0, 1, 4, 7, and 14 after nerve transection. Furthermore, we predicted the target genes of these microRNAs and analyzed the biological processes in which they were involved. The identified biological processes were consistent with the known time-frame of peripheral nerve injury and repair. Our data provide an important resource for further study of the role and regulation of microRNAs in peripheral nerve injury and regeneration.
LI ShiYing YU Bin WANG YongJun YAO DengBing ZHANG ZhanHu GU XiaoSong
