Abstract
BackgroundThe n6-methyladenosine (m6A) modification is present widely in mRNAs and long non-coding RNAs (lncRNAs), and is related to the occurrence and development of certain diseases. However, the role of m6A methylation in Clostridium perfringens type C infectious diarrhea remains unclear.MethodsHere, we treated intestinal porcine jejunum epithelial cells (IPEC-J2 cells) with Clostridium perfringens beta2 (CPB2) toxin to construct an in vitro model of Clostridium perfringens type C (C. perfringens type C) infectious diarrhea, and then used methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) to identify the methylation profiles of mRNAs and lncRNAs in IPEC-J2 cells.ResultsWe identified 6,413 peaks, representing 5,825 m6A-modified mRNAs and 433 modified lncRNAs, of which 4,356 m6A modified mRNAs and 221 m6A modified lncRNAs were significantly differential expressed between the control group and CPB2 group. The motif GGACU was enriched significantly in both the control group and the CPB2 group. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation analysis showed that the differentially methylated modified mRNAs were mainly enriched in Hippo signaling pathway and Wnt signaling pathway. In addition, the target genes of the differentially m6A modified lncRNAs were related to defense response to virus and immune response. For example, ENSSSCG00000042575, ENSSSCG00000048701 and ENSSSCG00000048785 might regulate the defense response to virus, immune and inflammatory response to resist the harmful effects of viruses on cells.ConclusionIn summary, this study established the m6A transcription profile of mRNAs and lncRNAs in IPEC-J2 cells treated by CPB2 toxin. Further analysis showed that m6A-modified RNAs were related to defense against viruses and immune response after CPB2 toxin treatment of the cells. Threem6A-modified lncRNAs, ENSSSCG00000042575, ENSSSCG00000048785 and ENSSSCG00000048701, were most likely to play a key role in CPB2 toxin-treated IPEC-J2 cells. The results provide a theoretical basis for further research on the role of m6A modification in piglet diarrhea.
Highlights
Bacterial diarrhea in piglets, which leads to a decline in piglet survival and seriously affects pig husbandry, is an urgent problem in pig production
IPEC-J2 cells were stimulated with C. perfringens beta2 (CPB2) (20 mg/mL) for 24 h, which was defined as the CPB2 group, and unstimulated IPEC-J2 cells were defined as the control group
To explore whether CPB2 toxin-treated cells were related to m6A methylation, the overall level of m6A methylation was determined
Summary
Bacterial diarrhea in piglets, which leads to a decline in piglet survival and seriously affects pig husbandry, is an urgent problem in pig production. Clostridium perfringens type C (C. perfringens type C) is one of the main pathogenic bacteria causing diarrhea in piglets [1]. C. perfringens type C attaches to the top of the villi of intestinal epithelial cells and multiplies in the chorionic basement membrane to produce C. perfringens beta (CPB2) toxin. The prevention and treatment of this diarrhea disease relies mainly on vaccines and antibiotics. These measures have reduced and controlled the occurrence of diarrhea to some extent, longterm use of antibiotics has led to increased bacterial resistance and resulted in food safety and human health concerns. Mutation of pathogenic bacteria is a continuous challenge for the research and development of vaccines, making the prevention and treatment of piglet diarrhea more difficult. The role of m6A methylation in Clostridium perfringens type C infectious diarrhea remains unclear
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