CULTURE SUPERNATANT OF Lactobacillus plantarum CRL759 ANTI-INFLAMMATORY EFFECT ON LPS-CHALLENGED HUMAN RETINAL PIGMENT EPITHELIUM CELLS

BARBARA I LAYUS; GRACIELA FONT DE VALDEZ; ANA V RODRIGUEZ

Buenos Aires

Congreso; JOINT MEETING OF BIOSCIENCE SOCIETIES - LIII ANNUAL MEETING OF ARGENTINE SOCIETY OF BIOCHEMISTRY AND MOLECULAR BIOLOGY (SAIB); 2017

CULTURE SUPERNATANT OF Lactobacillus plantarum CRL759 ANTI-INFLAMMATORY EFFECT ON LPS-CHALLENGED HUMAN RETINAL PIGMENT EPITHELIUM CELLS Barbara Ivana Layus Graciela Font; Ana Virginia RodriguezCentro de Referencia para Lactobacilos (CERELA)- CONICET. Chacabuco 145. CPA: (T4000ILC). Tucumán, Argentina/ E-mail: blayus@cerela.org.arSoluble factors produced by probiotic lactic acid bacteria (LAB) such as lactobacilli can reduce inflammatory and oxidative damage. Previous studies in our laboratory showed the immunomodulatory capacity of L. plantarum strains on ARPE-19 cells (human retinal pigment epithelium cell line). The aims of this study were to determine whether L. plantarum CRL759 supernatant (LplS) was able to modulate the inflammatory response and cells migration triggered by LPS on ARPE-19 cells.L. plantarum was cultured in DMEM medium at 37ºC, 5% CO2; LplS was obtained by filtration on 0.22µm membranes. The cytotoxicity of the LplS was determined by MTT assay. A sub- cytotoxic dose of LplS or dexamethasone were pre-incubated with 2.5x105 cells before LPS (50 µ/ml) challenge. Cytokines and nitric oxide (NO) produced by ARPE-19 cells were measured by flow cytometry and Griess assay respectively. Thiobarbituric acid reactive substances (TBARS) were determined as the measure of lipid peroxidation. Migration ability was evaluated using the assay of perpendicular scratch to the cell layer by ImageJ programe. Dexamethasone was used as anti-inflammatory control in all assays.A dilution of 1:2 of the LplS did not exert toxic effects on ARPE-19 cells after 24h of incubation. LPS increased pro-inflammatory mediators produced by cells. Results showed that levels of IL-6, IL-8, NO and TBARS were significantly lower (50, 71, 93 and 98% respectively) than the LPS stimulated cells. Anti-inflammatory effects of LplS were comparable to the one achieved by dexamethasone. Migration of LPS-stimulated cells with dexamethasone treatment showed significant decrease at 24h, but cells treated with LplS significantly increased migration at the same time. This is the first report about a probiotic effect on ARPE-19 inflammatory model.Key words: probiotic; inflammation; ARPE-19