Effects of lactic acid bacteria on inflammatory cytokines production by ARPE-19 cells

DIAZ, MA; GRIET, MILAGROS; DE MORENO, ALEJANDRA; GONZALEZ, SILVIA NELINA; RODRIGUEZ, VIRGINIA

Córdoba

Congreso; LII Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2016

SAIB

The anti-inflammatory capacity of probiotic lactic acid bacteria was extensively studied as therapy for inflammatory bowel diseases,but only two studies have been performed in ocular inflammatory disorders. The aim of this work was to investigate the capacity of 4strains ofLactobacillus plantarum(CRL 1081, CRL759, CRL 725, CRL 92) to modulate cytokines production of LPS-challengedARPE-19 cells (human retinal pigment epithelial cell line). First, bacteria were grown in MRS medium, showing that exponential growth phase was the optimal for co-culture with cells. Arpe-19 cells were co-incubated with bacterial strains at 10 and 20 MOI inDMEM medium for 6 hours at 37ºC. Then, the cells were stimulated with different LPS concentration (10, 50 and 100 µg/ml) for 24hours. IL-6, IL-8, IL-1β, TNF-α, IL-10 and IL-12p70 were detected in culture supernatants using flow cytometric bead arraytechnique. ARPE-19 cells treated with 10µg/ml LPS produced IL-6 (141.4 pg/ml) and IL-8 (335 pg/ml). Two strains exerted an antiinflammatoryeffect:L. plantarumCRL 725 at 20 MOI reduced the production of IL-6 and IL-8 by 25% andL. plantarumCRL 759 at 10MOI totally reverted the secretion of both cytokines induced by LPS in ARPE-19 cells. No IL-1β, IL-10, TNF-α and IL-12p70secretion was detected. Experimental assays to determine the mechanisms responsible for this effect are under development