Effect of Panax ginseng extract on S. aureus invasion into bovine mammary epithelial cells and cytokines production

BECCARIA C; SILVESTRINI P; SACCO S; LOVATTO M; RENNA MS; CALVINHO L; BARAVALLE C; DALLARD BE

Mar del Plata (Bs. As.)

Congreso; LXI Reunión Anual de la Sociedad Argentina de Investigación Clínica (SAIC), LXIV Reunión anual de la Sociedad Argentina de Inmunología (SAI) y XLVIII Reunión Anual de la Sociedad Argentina de Farmacología Experimental (SAFE).; 2016

Effect of Panax ginseng extract on Staphylococcus aureus invasion into bovine mammary epithelial cells and cytokines production Beccaria C1, Silvestrini P1, Sacco S1, Lovato MB, Renna MS1, Calvinho LF2, Baravalle C1, Dallard BE1. 1. Laboratorio de Biolog¨ªa Celular y Molecular Aplicada. Instituto de Ciencias Veterinarias del Litoral (ICIVET-Litoral) (UNL-CONICET). 2. EEA Rafaela, INTA. Panax ginseng extract (PGe) has been widely used as an herbal remedy for various disorders in humans and animals. The aim of this study was to evaluate the effect of PGe on S. aureus invasion into bovine mammary epithelial cells (MAC-T) and quantify the inflammatory response. The MAC-T cells viability treated with different concentrations of PGe (0, 0.5, 1 and 3 mg/ml) was measured by XTT assay. Then, supernatants from MAC-T cultured in 24-well plates with or without different concentrations of PGe were employed for cytokines ELISA test. For S. aureus invasion assays, MAC-T were cultured with PGe (0, 0.5, 1 and 3 mg/ml) before infecting with a S. aureus strain isolated from bovine mastitis, with a multiplicity of infection (MOI) 100:1 bacteria/cell. After challenged with S. aureus for 2 h the MAC-T were washed and treated with gentamicin for 2 h to eliminate extracellular bacteria. Supernatants were then collected and cultured to verify killing by gentamicin. Data were expressed as log10 of colony forming units (CFU)/ml of internalized bacteria. No cytotoxic effects of PGe on MAC-T viability were observed. The highest concentration of PGe (3 mg/ml) induced an increase in the IL-4 production compared with other concentrations and control after 24 h of treatment (p<0.01). The IL-1¦Â and IL-6 levels were below the lowest detectable test limit in all treatments at 24 h. According to CFU/ml recovered, PGe inhibited S. aureus invasion into MAC-T cells in a dose-dependent manner (0.5 to 3 mg/ml) ranging from 5,82 to 5,28 log10 CFU/ml respectively compared with control cells (6,08 log10) (p<0.01). Results demonstrated that PGe significantly inhibited S. aureus invasion into MAC-T cells and promoted IL-4 production (anti-inflammatory cytokine). In this way, PGe could modify the early S. aureus-host interaction in bovine mammary gland. The properties of PGe reported here may open new way for the development of novel prevention or treatment strategies for bovine mastitis. Keywords: Staphylococcus aureus; Panax ginseng; Bovine mammary epithelial cells; Invasio