Exogenous hydrogen sulfide prevents lipopolysaccharide-induced inflammation by blocking the TLR4/NF-κB pathway in MAC-T cells

Abstract

Mastitis impairs animal health and results in economic loss. Lipopolysaccharide (LPS) may cause immune response and inflammation in the bovine mammary gland. Hydrogen sulfide (H₂S) is the third gasotransmitter that acts as an anti-inflammation regulator in many cells. Despite the importance of H₂S in regulating inflammation, the effect and mechanism of exogenous H₂S on LPS-induced inflammation in bovine mammary epithelial cells are unknown. In the present study, with NaHS as a donor of H₂S, the bovine mammary epithelial cell line (MAC-T) was applied as an in vitro model to study the role of H₂S on LPS-induced MAC-T cells. The results verified that the cell viability was diminished by LPS but restored by exogenous H₂S at a physiologically relevant concentration (10 μM). Additionally, the production of H₂S was mitigated in the LPS-induced MAC-T cells. Meanwhile, exogenous H₂S decreased the intracellular ROS production and mRNA expression levels of the pro-inflammatory cytokines, TNF-α, IL-1β, IL-8, and IL-6. Furthermore, exogenous H₂S inhibited the mRNA expression of TLR4 and activation of NF-κB signaling pathway. In summary, exogenous H₂S exerts anti-inflammatory effects through attenuating oxidative stress and blocking the TLR4/NF-κB pathway in the LPS-induced bovine mammary epithelial cells. Our findings might clarify new prophylactic approaches for mastitis.

Keywords: Anti-inflammatory; Dairy mammary epithelial cells; Hydrogen sulfide; Lipopolysaccharide; NF-κB signaling pathways.