Eliglustat protects from damage caused by Shiga toxin type 2 in human renal tubular epithelial cells.

SÁNCHEZ DS; FISCHER SIGEL LK; BALESTRACCI A; IBARRA C; AMARAL MM; SILBERSTEIN C

Buenos Aires

Congreso; Reunión de las Sociedades de Biociencias. Modalidad virtual.; 2020

SAIC. SAI.

Shiga toxin-producing Escherichia coli is responsible for Hemolytic Uremic Syndrome (HUS), a cause of renal failure in children. We have previously shown that C-9 and Eliglustat (EG), inhibitors of glucosylceramide synthase and globotriaosylceramide (Gb3), prevent the cytotoxic effects of Shiga toxin type 2 (Stx2), in human cortical renal tubular epithelial cells (HRTEC) primary cultures and HK2 cell line. The aim of this work was to evaluate the efficacy of EG, elucidating EG treatments necessary to achieve total protection against Stx2 in HRTEC and HK2. Cells were incubated with Stx2 (1 ng/ml, 24 and 72 h) and pre-incubated with or without EG (1-500 nM, 6 and 24 h), followed by co-incubation with same dilutions of EG and Stx2 (24 and 72 h). Total number of cells stained with Hoechst was counted in microphotographs and compared with cell viability measured by neutral red uptake. Early and late apoptosis and necrosis was evaluated by annexin V/propidium iodide staining. Tubulogenesis was evaluated in HRTEC grown on matrigel. Treatment of cellswith Stx2 significantly decreased cell confluence and viability and the number of cells attached (p