PDF(Pubmed)
Abstract：
Rabies virus (RABV) is a neurotropic virus that causes fatal neurological disease, raising serious public health issues and attracting extensive attention in society. To elucidate the molecular mechanism of RABV-induced neuronal damage, we used hematoxylin-eosin staining, transmission electron microscopy, transcriptomics analysis, and immune response factor testing to investigate RABV-infected neurons. We successfully isolated the neurons from murine brains. The specificity of the isolated neurons was identified by a monoclonal antibody, and the viability of the neurons was 83.53-95.0%. We confirmed that RABV infection induced serious damage to the neurons according to histochemistry and transmission electron microscope (TEM) scanning. In addition, the transcriptomics analysis suggested that multiple genes related to the pyroptosis pathway were significantly upregulated, including gasdermin D (Gsdmd), Nlrp3, caspase-1, and IL-1β, as well as the chemokine genes Ccl2, Ccl3, Ccl4, Ccl5, Ccl7, Ccl12, and Cxcl10. We next verified this finding in the brains of mice infected with the rRC-HL, GX074, and challenge virus standard strain-24 (CVS-24) strains of RABV. Importantly, we found that the expression level of the Gsdmd protein was significantly upregulated in the neurons infected with different RABV strains and ranged from 691.1 to 5764.96 pg/mL, while the basal level of mock-infected neurons was less than 100 pg/mL. Taken together, our findings suggest that Gsdmd-induced pyroptosis is involved in the neuron damage caused by RABV infection.
摘要：
狂犬病病毒（RABV）是一种嗜神经病毒，可导致致命的神经系统疾病，引起了社会的广泛关注。阐明RABV诱导神经元损伤的分子机制,我们用苏木精-伊红染色,透射电子显微镜,转录组学分析，和免疫反应因子测试以研究RABV感染的神经元。我们成功地从鼠脑中分离出神经元。通过单克隆抗体鉴定分离的神经元的特异性，神经元的活力为83.53-95.0％。根据组织化学和透射电子显微镜（TEM）扫描，我们证实了RABV感染对神经元的严重损伤。此外,转录组学分析表明，与焦亡途径相关的多个基因显著上调，包括gasderminD(Gsdmd)，Nlrp3、caspase-1和IL-1β，以及趋化因子基因Ccl2，Ccl3，Ccl4，Ccl5，Ccl7，Ccl12和Cxcl10。接下来，我们在感染rRC-HL的小鼠的大脑中验证了这一发现，GX074和攻击RABV的病毒标准毒株-24(CVS-24)毒株。重要的是,我们发现Gsdmd蛋白的表达水平在不同RABV株感染的神经元中显著上调，范围从691.1到5764.96pg/mL，而模拟感染神经元的基础水平低于100pg/mL。一起来看,我们的发现表明Gsdmd诱导的焦亡参与了RABV感染引起的神经元损伤。
