The Mirna Profiles Analysis Of Murine Spinal Cord Infected With Rabies Virus And The Preliminary Study Of The Interaction

Rabies virus is a member of the Lyssavirus genus of the Rhabdoviridae family. It is a highly neurotropic virus that can cause fatal infections of warm-blooded animals. At present, the pathogenesis of rabies virus has not been fully understood. For without treatment, prevention and control by vaccination only is the situation. Therefore, to study the pathogenic mechanism is particularly important by different ways.MicroRNA (miRNA) is a class of endogenous small molecule with a length of about21-23nt non-coding single-strand RNA. It is an important class of gene regulation by completely or incompletely paired with the target gene, in order to play a role in degradation mRNA or inhibition of mRNA translation. miRNA plays an important role in expression and regulation of genes, cell differentiation, cellular apoptosis and antiviral defense.In this study, we constructed a small RNA library and sequencing analysis of partial small RNA in the RNA library extracting from spinal cord tissue of normal mouse or mouse infected with rabies virus GX01. We acquried a miRNA expression profile of mouse brain infected with rabies virus GX01. The miRNAs which have up-regulated expression more than three folds contain miR-34a-3p, miR-155-5p, miR-223-3p and miR-223-5p. However, the miRNA with three folds down-regulated expression was miR-193b-5p only. Then we analysised the KEGG pathway via predicting the five differentially expressed miRNA target genes, and further139new miRNAs in total were verified to relate with rabies virus infection by computer analysis. In addition, we validated miR-34a-3p, miR-155-5p and miR-223-3p three differentially expressed miRNA by real-time qRCR, showing that differential expression folds upward trend were the similar with miR-155-5p and miR-223-3p. However, the miR-34a-3p was not consistent, initially speculating that the results may be related to detection methods.The three miRNAs miR-101c, miR-155, miR-223were selected to construct a recombinant expressing plasmid pEGFP-C1-miR-101c, pEGFP-C1-miR-155, pEGFP-C1-miR-223, and these miRNAs were successfully expressed in BHK cells, based on obtaining the target genes from miRBasce and amplified miRNA primary transcripts including its precursor. At the same time, to understand how miRNAs play antiviral activity with their targeting gene, we explored the interaction between the three recombinant expression plasmids and rabies virus GX01. The results showed that the three recombinant expression plasmids could reduce viral titer to a certain extent and suppressed multipility rate of rabies virus. N gene transcription of rabies virus was not significantly changed post infected12h and24h, while its transcription increased in different degrees in infected cells during48h to72h. However, at the level of protein expression, rabies virus N protein expression was not significantly changed. miRNA expression for the impact of rabies virus proliferation rate and its interaction with the host remains to be a further study.