Inhibitory effect of an ethnoveterinary oil and water formulation on LTA-induced mastitis via TLR2/NF-κB signaling in mammary epithelial cells

Abstract

Mastitis, an inflammatory disease, becoming a problematic condition in the dairy industry, which causes severe economic loss. It is primarily infected by Staphylococcus aureus (S. aureus). Current conventional therapies for mastitis often lead to antibiotics resistance. To address this issue, a plant based ethnoveterinary medicine approach is being explored as a potential treatment for this condition. The aim of the present study was to evaluate the anti-inflammatory and anti-mastitis effect of oil based (EO) and water based (EW) ethno veterinary formulations in Lipoteichoic acid (LTA) stimulated HC11 cells. LTA induced inflammatory model in HC11 mouse mammary epithelial cells were used in this study. The inflammatory genes expressions such as inducible nitric oxide synthase (iNOS), Toll like receptor 2 (TLR2), and NF-κB p65 were analysed by RT-PCR analysis. The antioxidant enzymes and inflammatory mediators were analysed using ELISA method. NAGase activity was analysed by using colorimetric method. Reactive oxygen species (ROS) generation was assessed by fluorescent microscopy method. Results revealed that EO inhibited the production of nitric oxide (NO), Lipid perioxidation and the gene expression of iNOS, TLR2, and NF-κB p65 in the LTA-induced HC11 cells. Moreover, the DCFH-DA staining showed that EO enhances the endogenous antioxidant enzymes activity and mitigated the oxidative stress and ROS generation. Also, results showed that the treatment with EO possesses potent anti-mastitis effect by inhibiting NAGase activity. These results indicated that EO exhibits its anti-inflammatory and anti-mastitis effects mainly through attenuating TLR2 and NF-κB signaling pathway. Hence, EO could be a potential therapeutic agent against mastitis.