Integrative Bioinformatics Indentification Of The Autophagic Pathway-Associated Mirna-Mrna Networks In Raw264.7 Macrophage Cells Infected With Increment Omp25 Brucella Melitensis

INFLAMMATION(2020)

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Abstract
Brucellosis is a zoonotic infectious disease caused by Brucella infection. Outer membrane protein 25 (Omp25) is closely related to the virulence and immunogenicity of Brucella. However, the molecular mechanism of Omp25 affecting Brucella-mediated macrophage autophagy remains unclear. Our previous study reported that four miRNAs (the upregulation of mmu-miR-146a-5p and mmu-miR-155-5p and downregulation of mmu-miR-149-3p and mmu-miR-5126) were confirmed and revealed the differentially expressed genes (DEGs) profile in RAW264.7 macrophage cells infected with Brucella melitensis Omp25 deletion mutant (Delta Omp25 B. melitensis). Here, we predicted the target genes of the four miRNAs by TargetScan, miRanda, and PicTar. GO and KEGG were used for functional enrichment analysis of DEGs profile to reveal the autophagic pathway-associated genes. The overlapped genes, which drawn the autophagic pathway-associated miRNA-mRNA networks by cytoscape software, were identified by intersecting with the predicted target genes and autophagic pathway-associated DEGs. qRT-PCR was performed to validate the mRNAs of networks. The results showed that the autophagic pathway-associated networks of mmu-miR-149-3p-Ptpn5, mmu-miR-149-3p-Ppp2r3c, and mmu-miR-146a-5p-Dusp16 were identified in RAW264.7 macrophage cells infected with Delta Omp25 B. melitensis. Our findings are of great significance in elucidating the function of Omp25, revealing the infection mechanism of Brucella and prophylaxising and treating brucellosis.
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Key words
Brucella,Omp25,RAW264.7,autophagy,miRNA-mRNA network
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