Metabolomic Profiles of Bovine Mammary Epithelial Cells Stimulated by Lipopolysaccharide

Abstract

Bovine mammary epithelial cells (bMECs) are the main cells of the dairy cow mammary gland. In addition to their role in milk production, they are effector cells of mammary immunity. However, there is little information about changes in metabolites of bMECs when stimulated by lipopolysaccharide (LPS). This study describes a metabolomics analysis of the LPS-stimulated bMECs to provide a basis for the identification of potential diagnostic screening biomarkers and possible treatments for bovine mammary gland inflammation. In the present study, bMECs were challenged with 500 ng/mL LPS and samples were taken at 0 h, 12 h and 24 h post stimulation. Metabolic changes were investigated using high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF MS) with univariate and multivariate statistical analyses. Clustering and metabolic pathway changes were established by MetaboAnalyst. Sixty-three differential metabolites were identified, including glycerophosphocholine, glycerol-3-phosphate, L-carnitine, L-aspartate, glutathione, prostaglandin G2, α-linolenic acid and linoleic acid. They were mainly involved in eight pathways, including D-glutamine and D-glutamic acid metabolism; linoleic acid metabolism; α-linolenic metabolism; and phospholipid metabolism. The results suggest that bMECs are able to regulate pro-inflammatory, anti-inflammatory, antioxidation and energy-producing related metabolites through lipid, antioxidation and energy metabolism in response to inflammatory stimuli.

Alterations in immune and antioxidant gene networks by gamma-d-glutamyl-meso-diaminopimelic acid in bovine mammary epithelial cells are attenuated by in vitro supply of methionine and arginine

“…Bovine mammary epithelial cells contribute to the innate immune response during infection of the mammary gland by recognizing pathogens through PRR (Ibeagha-Awemu et al, 2008;Huang et al, 2019). The NOD-like receptors are cytosolic PRR, which compensate for and synergize with other PRR, such as toll-like receptors (TLR) during infection (Cardenas et al, 2011).…”

Effects of Peptidoglycan, Lipoteichoic Acid and Lipopolysaccharide on Inflammation, Proliferation and Milk Fat Synthesis in Bovine Mammary Epithelial Cells

“…The construction of mastitis model of cows was frequently based on 1 μg/mL LPS stimulation for BMECs in vitro in many studies [ 33 , 34 , 35 , 36 , 37 ]. There were some studies which reported that a lower concentration of LPS, such as 0.1 [ 38 , 39 ], 0.2 [ 40 , 41 ], 0.5 [ 42 ] μg/mL and so on, could also induce inflammatory responses of BMECs. In the present study, 0.01 μg/mL LPS only significantly promoted the gene expression of IL-6 and IL-8 .…”

Ketogenic Diets Induced Glucose Intolerance and Lipid Accumulation in Mice with Alterations in Gut Microbiota and Metabolites

“…The raw data were converted into mzXML format using Proteo Wizard software, and peak alignment, retention time correction, and peak area extraction were performed using the XCMS program. Details of the data processing and analysis were performed as described previously ( 54 ).…”