Abstract
Current knowledge on the role of microRNAs (miRNAs) in rabbit hemorrhagic disease virus (RHDV) infection and the pathogenesis of rabbit hemorrhagic disease (RHD) is still limited. RHDV replicates in the liver, causing hepatic necrosis and liver failure. MiRNAs are a class of short RNA molecules, and their expression profiles vary over the course of diseases, both in the tissue environment and in the bloodstream. This paper evaluates the expression of miRNAs in the liver tissue (ocu-miR-122-5p, ocu-miR-155-5p, and ocu-miR-16b-5p) and serum (ocu-miR-122-5p) of rabbits experimentally infected with RHDV. The expression levels of ocu-miR-122-5p, ocu-miR-155-5p, and ocu-miR-16b-5p in liver tissue were determined using reverse transcription quantitative real-time PCR (RT-qPCR), and the expression level of circulating ocu-miR-122-5p was established using droplet digital PCR (ddPCR). The expression levels of ocu-miR-155-5p and ocu-miR-16b-5p were significantly higher in the infected rabbits compared to the healthy rabbits (a fold-change of 5.8 and 2.5, respectively). The expression of ocu-miR-122-5p was not significantly different in the liver tissue from the infected rabbits compared to the healthy rabbits (p = 0.990), while the absolute expression level of the circulating ocu-miR-122-5p was significantly higher in the infected rabbits than in the healthy rabbits (p < 0.0001). Furthermore, a functional analysis showed that ocu-miR-155-5p, ocu-miR-16b-5p, and ocu-miR-122-5p can regulate the expression of genes involved in processes correlated with acute liver failure (ALF) in rabbits. Search tool for the retrieval of interacting genes/proteins (STRING) analysis showed that the potential target genes of the three selected miRNAs may interact with each other in different pathways. The results indicate the roles of these miRNAs in RHDV infection and over the course of RHD and may reflect hepatic inflammation and impairment/dysfunction in RHD.
Highlights
Rabbit hemorrhagic disease virus (RHDV), genotype GI.1, belongs to the family Caliciviridae, genusLagovirus [1], and is an etiological agent of rabbit hemorrhagic disease (RHD)
The present results evaluate the expression of miRNAs in the liver tissue and serum of rabbits experimentally infected with rabbit hemorrhagic disease virus (RHDV)
Infected rabbits were divided into groups to assess if there was a relationship between the expression level of ocu-miR-122-5p in the serum and the survival time of the animals
Summary
Rabbit hemorrhagic disease virus (RHDV), genotype GI., belongs to the family Caliciviridae, genusLagovirus [1], and is an etiological agent of rabbit hemorrhagic disease (RHD). Phylogenetic analysis showed a division of the 105 strains Lagovirus europaeus into seven groups, determined by their chronology, geographical location, and evolutionary events, such as mutation and recombinations [3]. This disease affects rabbit populations worldwide, and more than 30 years have passed since the first outbreak of rabbit hemorrhagic disease, the mechanisms underlying the pathogenesis of the disease are not yet fully understood. RHD-related death can be caused directly by systemic hemorrhagic diathesis and disseminated intravascular coagulation (DIC) [4], likely due to liver cell damage from apoptosis induced by RHDV [8,14].
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