■柯萨奇病毒A6(CV-A6)已成为主要的流行株,口蹄疫(HFMD)。CV-A6感染可导致严重的临床表现,包括脑炎,脑膜炎,和可能危及生命的中枢神经系统疾病。我们之前的研究结果表明,感染CV-A6的新生小鼠表现出肢体无力,瘫痪最终屈服于死亡。然而,CV-A6诱导的神经系统损伤的潜在机制仍然难以捉摸。许多报道强调了miRNA在各种病毒感染中的关键作用。
■使用分别建立的小鼠感染组和对照组,在CV-A6转染前后创建脑组织的miRNA谱,随后进行实验验证,预测,并对结果进行分析。
■感染后2天(dpi),4dpi,和2dpivs4dpi,为了验证目的,我们使用qRT-PCR分别鉴定了175,198和78个显著差异表达的miRNA.随后,我们预测了这些差异表达miRNA的靶基因,并通过GO(基因本体论)富集分析和KEGG(京都基因和基因组百科全书)富集分析确定了它们的潜在靶标。最后,我们通过双荧光素酶实验验证了miRNA-mRNA配对,同时通过WesternBlotting分析证实了靶基因的功能富集.
■这项研究的结果表明,转录调控,神经元坏死,促炎细胞因子释放,和抗病毒免疫都与CV-A6感染小鼠中枢神经系统损伤的发病机理有关。CV-A6感染引起的脑损伤可能涉及多种途径,包括神经胶质细胞激活,神经元坏死,突触破坏,神经系统的退行性疾病。它甚至可以破坏血脑屏障,导致中枢神经系统损伤。在这项研究中发现的失调的miRNA和信号通路为进一步研究CV-A6的发病机理提供了有价值的见解。
UNASSIGNED: Coxsackievirus A6 (CV-A6) has emerged as the predominant epidemic strain responsible for hand, foot and mouth disease (HFMD). CV-A6 infection can result in severe clinical manifestations, including encephalitis, meningitis, and potentially life-threatening central nervous system disorders. Our previous research findings demonstrated that neonatal mice infected with CV-A6 exhibited limb weakness, paralysis, and ultimately succumbed to death. However, the underlying mechanism of CV-A6-induced nervous system injury remains elusive. Numerous reports have highlighted the pivotal role of miRNAs in various viral infections.
UNASSIGNED: Separately established infection and control groups of mice were used to create
miRNA profiles of the brain tissues before and after CV-A6 transfection, followed by experimental verification, prediction, and analysis of the results.
UNASSIGNED: At 2 days post-infection (dpi), 4 dpi, and 2dpi vs 4dpi, we identified 175, 198 and 78 significantly differentially expressed miRNAs respectively using qRT-PCR for validation purposes. Subsequently, we predicted target genes of these differentially expressed miRNAs and determined their potential targets through GO (Gene Ontology) enrichment analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis. Finally, we verified the
miRNA-mRNA pairing via double luciferase experiments while confirming functional enrichment of target genes through Western Blotting analyses.
UNASSIGNED: The results from this study suggest that transcriptional regulation, neuronal necrosis, pro-inflammatory cytokine release, and antiviral immunity are all implicated in the pathogenesis of central nervous system injury in mice infected with CV-A6. Brain injury resulting from CV-A6 infection may involve multiple pathways, including glial cell activation, neuronal necrosis, synaptic destruction, degenerative diseases of the nervous system. It can even encompass destruction of the blood-brain barrier, leading to central nervous system injury. The dysregulated miRNAs and signaling pathways discovered in this study provide valuable insights for further investigations into the pathogenesis of CV-A6.