Study on the mechanism of miRNAs on liver injury in the condition of Protoscocephalus alveolarus transhepatic portal vein infection

Abstract

Objective: To explore the role of miRNA in liver damage caused by Echinococcus multilocularis infection.

Methods: Six female C57BL mice were randomly divided into two groups, the control group and the infection group. Mice in the control group were injected with 100 μL PBS through the hepatic portal vein, and mice in the infection group were infected with E. multilocularis via the hepatic portal vein to establish a mouse model of infection. Small RNA sequencing was performed for detecting the expression of miRNAs in the liver of mice infected with 2000 E. multilocularis after 3 months of infection, screen out miRNAs related to liver damage, and verify by RT-PCR.

Results: Seventy-one differentially expressed miRNAs were found in the liver in comparison with control, and a total of 36 mouse miRNAs with |FC| >0.585 were screened out, respectively. In addition, Targetscan (V5.0) and miRanda (v3.3a) software were used to predict differential miRNAs target genes and functional enrichment of target genes. Functional annotation showed that "cytokine-cytokine interaction," "positive regulation of cytokine production," "inflammatory response," and "leukocyte activation" were enriched in the liver of E. multilocularis-infected mice. Moreover, the pathways "human cytomegalovirus infection," "cysteine and methionine metabolism," "Notch signaling pathway," and "ferroptosis" were involved in liver disease. Furthermore, four miRNAs (mmu-miR-30e-3p, mmu-miR-203-3p, mmu-miR-125b-5p, and mmu-miR-30c-2-3p) related to liver injury were screened and verified.

Conclusion: This study revealed that the expression profiling of miRNAs in the livers was changed after E. multilocularis infection, and improved our understanding of the transcriptomic landscape of hepatic echinococcosis in mice.

Keywords: Echinococcus multilocularis; liver injury; miRNA.

Figure 1

Identification of mouse models of infection. (A) Imaging of Echinococcus multilocularis. (B) B‐ultrasound detection of the liver in mice. (C−F) Changes of E. multilocularis‐specific IgG, IgM, IgA, and IgE antibodies. (G−J) Changes of total IgG, IgM, IgA, and IgE antibodies. *p < .05, **p < .01, ***p < .005.

Figure 2

Echinococcus multilocularis infection causes liver damage. (A) The HE staining of mouse liver. (B) The Masson staining of mouse liver. (C) The level of AST and ALT in serum. *p < .05, **p < .01. The red arrows indicate inflammatory cells, black arrows indicate germinal layer, and green arrows indicate vesicle. HE, hematoxylin and eosin.

Figure 3

Differential expression of miRNAs. (A) Venn diagrams of detected miRNAs. (B) Differential expressed miRNAs in different groups. (C) Heat map of differential expression miRNAs. (D) Volcano plot of miRNAs.

Figure 4

Functional enrichment of miRNA target genes. (A) GO enrichment barplot. (B) KEGG enrichment scatterplot. GO, gene ontology; KEGG, Kyoto encyclopedia of genes and genomes.

Figure 5

Validation of liver injury‐related miRNAs and target genes by RT‐qPCR. (A) Change of log2 (fold change) and fold change of differential expression gene. (B) The level of liver injury‐related miRNAs expression. (C) The level of mRNA expression. *p < .05, **p < .01, ***p < .001.