| Toxoplasma gondii is one of the most successful intracellular protozoa in the world and can infect most warm-blooded animals.Intestinal flora can enhance metabolic function,resist pathogens,improve host immunity,and directly or indirectly affect most physiological functions of animals through these basic functions.Meanwhile,liver is also an important target organ for Toxoplasma infection,and intestinal flora can participate in and influence the pathogenesis of disease through the gut-liver axis and play an important role in liver disease.However,few studies have investigated the effects of toxoplasma infection at different times and doses on the transcriptome expression profile of liver and the relationship between toxoplasma infection and intestinal flora.Therefore,in this study,20 and 10 toxoplasma cysts were gavaged to mice,which were divided into high and low doses for gavage infection.Mouse liver samples were collected for mRNA and small RNA transcriptome sequencing,and cecal contents were collected for metagenomic sequencing,and the three sequencing results were analyzed.To elucidate the effects of Toxoplasma gondii infection on intestinal microflora diversity and function and gene expression and transcriptional regulation in liver tissue.The results are as follows:(1)There were the most differentially expressed genes in the liver mRNA transcriptome of mice in the high-dose infection group during the acute infection stage,including 2622 upregulated genes and 1737 down-regulated genes.However,there were fewer up-regulated and down-regulated genes in the low-dose infection group compared with the control group at acute infection stage(66 and 73 genes,respectively.In chronic infection stage,the differential gene expression level in liver of mice decreased in both high and low dose infection groups.In addition,due to the different dose of infection,there are more different genes between the acute and chronic stage in the high-dose infection group than in the low-dose infection group.In the low-dose infection group,GO and KEGG enrichment analysis showed that the most annotated items and pathways were adipocyte differentiation and PPAR signaling pathway,respectively.GO and KEGG enrichment analysis in the high-dose infection group showed that the most annotated entries and pathways were WBC-cell adhesion and cytokine-cytokine receptor interaction,respectively.(2)R package DESeq2 was used for mi RNA difference analysis.Differential mirnas were found only in the liver of mice infected with Toxoplasma gondii at high doses.Among them,the number of differential mirnas in acute infection stage was the largest,with 53 mirnas,while only 5 mirnas existed in chronic infection stage.In acute infection,GO and KEGG enrichment analysis showed that the most annotated entries and pathways were dendrite development and MAPK signaling pathway,respectively.In chronic infections,GO and KEGG enrichment analyses showed that the most annotated entries and pathways were transcriptional dysregulation in osteogenesis and cancer,respectively.(3)A total of 156 SGB were obtained through metagenomic data assembly and species annotation was carried out.A total of 145 species were annotated.These species are classified into 9 different phyla,31 families and 78 genera.The α diversity of intestinal flora at phylum,family and species level increased,and there were significant inter-group differences in species βdiversity among groups.The abundance analysis showed that the intestinal flora abundance in the infection group was lower than that in the control group,and 9 characteristic families of acute infection period and 8 characteristic families of chronic infection period were screened out during the acute infection period.Functional analysis showed that the KEGG pathway enriched most significantly in amino acid metabolism during acute infection and energy metabolism during chronic infection.CAZy database annotation analysis showed that GH43 was the most abundant carbohydrate family in acute infection stage and GT4 in chronic stage.COG analysis showed that the replication,recombination and repair functions of homologous proteins were significantly enriched in both acute and chronic phases,except for unknown functions.In this study,transcriptome,small RNA and metagenomic sequencing were used to determine the differential gene expression and transcriptional regulation of intestinal flora and liver in healthy mice and Toxoplasma gondii infected mice.To elucidate the relationship between the diversity and function of intestinal flora and gene expression and transcriptional regulation in liver tissue.It provides supplementary data for the transcriptome,small RNA and intestinal microflora metagenome of mice,and provides reference for the study of toxoplasma pathogenic mechanism and the prevention and treatment of toxoplasmosis. |
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