Allicin Alleviated LPS-Induced Mastitis via the TLR4/NF-κB Signaling Pathway in Bovine Mammary Epithelial Cells

Int J Mol Sci. 2023 Feb 14;24(4):3805. doi: 10.3390/ijms24043805.

Abstract

Dairy farming is the most important economic activity in animal husbandry. Mastitis is the most common disease in dairy cattle and has a significant impact on milk quality and yield. The natural extract allicin, which is the main active ingredient of the sulfur-containing organic compounds in garlic, has anti-inflammatory, anticancer, antioxidant, and antibacterial properties; however, the specific mechanism underlying its effect on mastitis in dairy cows needs to be determined. Therefore, in this study, whether allicin can reduce lipopolysaccharide (LPS)-induced inflammation in the mammary epithelium of dairy cows was investigated. A cellular model of mammary inflammation was established by pretreating bovine mammary epithelial cells (MAC-T) with 10 µg/mL LPS, and the cultures were then treated with varying concentrations of allicin (0, 1, 2.5, 5, and 7.5 µM) added to the culture medium. MAC-T cells were examined using RT-qPCR and Western blotting to determine the effect of allicin. Subsequently, the level of phosphorylated nuclear factor kappa-B (NF-κB) was measured to further explore the mechanism underlying the effect of allicin on bovine mammary epithelial cell inflammation. Treatment with 2.5 µM allicin considerably decreased the LPS-induced increase in the levels of the inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) and inhibited activation of the NOD-like receptor protein 3 (NLRP3) inflammasome in cow mammary epithelial cells. Further research revealed that allicin also inhibited the phosphorylation of inhibitors of nuclear factor kappa-B-α (IκB-α) and NF-κB p65. In mice, LPS-induced mastitis was also ameliorated by allicin. Therefore, we hypothesize that allicin alleviated LPS-induced inflammation in the mammary epithelial cells of cows probably by affecting the TLR4/NF-κB signaling pathway. Allicin will likely become an alternative to antibiotics for the treatment of mastitis in cows.

Keywords: MAC-T; NF-κB; NLRP3; allicin; anti-inflammation; mastitis.

MeSH terms

  • Animals
  • Cattle
  • Disulfides* / therapeutic use
  • Epithelial Cells / metabolism
  • Female
  • Inflammation / metabolism
  • Interleukin-6 / metabolism
  • Lipopolysaccharides
  • Mastitis, Bovine* / drug therapy
  • Mice
  • NF-kappa B* / metabolism
  • Signal Transduction
  • Sulfinic Acids* / therapeutic use
  • Toll-Like Receptor 4 / metabolism

Substances

  • allicin
  • Disulfides
  • Interleukin-6
  • Lipopolysaccharides
  • NF-kappa B
  • Sulfinic Acids
  • TLR4 protein, human
  • Tlr4 protein, mouse
  • Toll-Like Receptor 4