Study of Mrp2 impairment induced by fructose in primary culture of hepatocytes. Preliminary data

BAROSSO, ISMAEL R; MEDEOT, ANABELA C; SCHUCK, VIRGINIA S; ANDERMATTEN, ROMINA B; CIRIACI, N; SANCHEZ POZZI, ENRIQUE J.

Congreso; Reunión anual Safis ALACF 2021; 2021

Sociedad Argentina de Fisiología

The increment consumption of fructose in the world diet contributes to the rise in total calorie intake and is related to an increase in the incidence of the Metabolic Syndrome (MS). Previously, we demonstrated that administration of 10% fructose in drinking water over 8 weeks to normal rats, a model of MS, reduced bile flow and decreased the expression of the canalicular transportersMrp2 (Multidrug resistance associated protein 2) and Bsep (bile salt export pump). To study the signaling pathways involved in fructose actions, we develop a cellular model to evaluate its effects on Mrp2. Methods: The study was carried out in sandwich‑cultured rat hepatocytes. Cells were treated with fructose (1-10-24 mM) for 3 days. LDH (lactate dehydrogenase) release in the media was measured by a detention kit (Wienner Lab). Mrp2 activity was evaluated by secretion of fluorescent methylfluorescein (GMF) using the index BEI (biliary excretion index). For that, treatments were performed duplicated, 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%. Results: The release of LDH to the medium (%Control) was increased after cells treatment with Fructose 10 mM (357*±32) and Fructose 24 mM (574*±32) increased the release of LDH to the medium. n=4 (*p