Abstract

Bovine mastitis is an inflammatory response of mammary glands caused by pathogenic microorganisms such as Escherichia coli (E. coli). As a key virulence factor of E. coli, lipopolysaccharide (LPS) triggers innate immune responses via activation of the toll-like-receptor 4 (TLR4) signaling pathway. However, the molecular regulatory network of LPS-induced bovine mastitis has yet to be fully mapped. In this study, bovine mammary epithelial cell lines MAC-T were exposed to LPS for 0, 6 and 12 h to assess the expression profiles of long non-coding RNAs (lncRNAs) using RNA-seq. Differentially expressed lncRNAs (DElncRNAs) were filtered out of the raw data for subsequent analyses. A total of 2,257 lncRNAs, including 210 annotated and 2047 novel lncRNAs were detected in all samples. A large proportion of lncRNAs were present in a high abundance, and 112 DElncRNAs were screened out at different time points. Compared with 0 h, there were 22 up- and 25 down-regulated lncRNAs in the 6 h of post-infection (hpi) group, and 27 up- and 22 down-regulated lncRNAs in the 12 hpi group. Compared with the 6 hpi group, 32 lncRNAs were up-regulated and 25 lncRNAs were down-regulated in the 12 hpi group. These DElncRNAs are involved in the regulation of a variety of immune-related processes including inflammatory responses bMECs exposed to LPS. Furthermore, lncRNA TCONS_00039271 and TCONS_00139850 were respectively significance down- and up-regulated, and their target genes involve in regulating inflammation-related signaling pathways (i.e.,Notch, NF-κB, MAPK, PI3K-Akt and mTOR signaling pathway), thereby regulating the occurrence and development of E. coli mastitis. This study provides a resource for lncRNA research on the molecular regulation of bovine mastitis

Highlights

  • Mastitis is one of the most common diseases in dairy cows worldwide, causing damage to mammary tissue, reduces milk production, decreases the quality of dairy products, and increases mortality and culling rates [1, 2]

  • Novel long non-coding RNAs (lncRNAs) were classified as intergenic lncRNA, antisense lncRNA, and sense overlapping (41.3%) (Figure 1B)

  • The results indicated that these DElncRNA may play an important role in the regulation of the inflammatory response following LPS treatment of bovine mammary epithelial cells (bMECs)

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Summary

Introduction

Mastitis is one of the most common diseases in dairy cows worldwide, causing damage to mammary tissue, reduces milk production, decreases the quality of dairy products, and increases mortality and culling rates [1, 2]. When E. coli infects host cells, lipopolysaccharide (LPS, a pathogenassociated molecular pattern) [3, 9] is recognized by toll-likereceptor 4 (TLR4) which triggers immune responses, acting as an early signal of pathogenic microbial infection [8, 10]. This leads to activation of the nuclear factor kappa B (NF-κB) signaling pathway and induction of the expression of several proinflammatory genes, such as interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) [11]. Few studies on the regulatory mechanisms in dairy cows have been reported in the literature, and this is a critical shortcoming

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