decreased mrp2 expression by fructose in the primary culture model in sandwich-cultured rat hepatocytes

BAROSSO ISMAEL RICARDO; MEDEOT ANABELA CAROLINA; SCHUCK VIRGINIA SOLEDAD; AGUSTINA FAZZI; HILLOTE GERALDINE LUCIA; SALAS GIMENA; SANCHEZ POZZI ENRIQUE JUAN

BUENOS AIRES

Congreso; CONGRESO ANUAL DE FISIOLOGIA; 2023

SAFIS

The increase in fructose consumption in the global diet contributes to an increase in total caloric intake and it is related to anincrease in the incidence of Metabolic Syndrome (MS). Previously, we demonstrated that administration of 10% fructose indrinking water for 8 weeks to normal rats, a model of MS, reduced bile flow and decreased the expression of canaliculartransporters Mrp2 (multidrug resistance-associated protein 2). and Bsep (export of bile salts bomb). Aim: To develop a model tostudy the signaling pathways involved in the actions of fructose, we evaluated whether incubation of a primary culture of rathepatocytes in collagen sandwich configuration (SCRHs) with fructose can mimic the effect on Mrp2 observed in vivo. Methods:Once the hepatocytes were polarized, they were treated with 22 mM fructose for 24 hours. LDH (lactate dehydrogenase)release in the media was measured by a detention kit (Wienner Lab). Mrp2 activity was evaluated by secretion of fluorescentglutathione methylfluorescein (GMF) using the index BEI (biliary excretion index). For that, treatments were performedduplicated, in one the experiment was performed with standard buffer whereas in the other a Ca2+/Mg2+ free buffer was used.The BEI of GMF was calculated as: BEI = (fluorescenceCa2+/Mg2+ - fluorescenceCa2+/Mg2+ -free) / fluorescence Ca2+/Mg2+x100%. Mrp2 expression was evaluated by SDS-polyacrylamide gel electrophoresis and transfer to PVDF membranes of cellhomogenates from both groups using primary antibodies directed against Mrp2. Results: (Average ± SEM). Fructose 24 mM didnot affect LDH (C: 100±5%; F: 105±3%; n=3) and produced a decrease in BEI by 60% with respect to the Control (C: 24±4; F: 9±4;n=4; *p