Transcriptome sequencing reveals fibrotic associated-genes involved in bovine mammary fibroblasts with Staphylococcus...

Bovine mammary fibrosis represents a considerable health problem of cows, primarily indicated by lactation failure. Staphylococcus aureus (S. aureus) can cause mammary damage, this multifactorial disease necessitates to identify how and to what extent molecular pathogen defense mechanisms prevent bacterial infections in bovine mammary gland. In this study, we have aimed to determine the transcriptional responses in bovine mammary fibroblasts (BMFBs) induced by S. aureus using bioinformatics analysis to determine whether mRNA expression profile changes between BMFBs activation and quiescence. Established primary BMFBs obtained from healthy Holstein bovine were induced 10⁶ CFU/mL heat-inactivated S. aureus and total RNA was isolated 6 h after treatment. The 574 DEGs were involved in gene ontology (GO) that were immune response, apoptotic process, extracellular region, receptor binding, endopeptidase activity and protein kinase activity et al. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, distinct pathway contained signaling molecules common to various inflammatory and fibrotic pathways were Pathways in cancer, Cytokine-cytokine receptor interaction, PI3K-Akt signaling pathway, TNF signaling pathway, MAPK signaling pathway and Toll-like receptor signaling pathway. The BMFBs was treated with heat-inactivated S. aureus (10⁶ CFU/mL) and also with pharmacological inhibitors of ERK1/2, P38 MAPK and JNK. The MMP-2 activity were examined gelatin zymography, MMP-2, TIMP-1, -2 and PLAU/PAI-1 protein expression were examined in vitro by western blot. The MMP-2 activity was significantly inhibited by simultaneous inhibition of ERK1/2, P38 MAPK and JNK, and MMP-2, TIMP-1,-2 and PLAU/PAI-1 protein expression were significantly decreased by inhibiting ERK1/2, P38 MAPK or JNK. This suggested a crosstalk between the ERK1/2, P38 MAPK or JNK signaling pathways in regulating extracellular matrix metabolism in the BMFBs with S. aureus. Our study complement our initial study on S. aureus-induced responses by fibrosis-associated genes in BMFBs. This may lead to development of novel therapeutic targets to control bovine mammary fibrosis induced by S. aureus.