NADPH oxidase participates in the cholestatic alterations induced by estradiol 17ß-D-glucuronide downstream MEK-ERK 1/2 kinases.

SALAS, GIMENA; MEDEOT, ANABELA CAROLINA; SCHUCK, VIRGINIA SOLEDAD; ANDERMATTEN, ROMINA B.; MISZCZUK, GISEL S.; CIRIACI, NADIA; RAZORI, M. VALERIA; BASIGLIO, CECILIA L; CROCENZI, FERNANDO A.

online

Congreso; REUNIÓN DE SOCIEDADES DE BIOCIENCIAS 2021; 2021

SAIC - SAI - AAFE - NANOMED-AR

We have previously demonstrated that NADPH oxidase (NOX)-gen- erated reactive oxygen species (ROS) are involved in the impair- ment of canalicular secretion induced by estradiol 17β-D-glucuro- nide (E17G) [Physiological Mini Reviews 12 (Special Edition): 019, 2019], and that NOX is in the same pathway that MEK-ERK 1⁄2 [Me- dicina (Bs. As.), 80 (supl V): 67, 2020]. Now, we intended to locate NOX into this pathway. To achieve this goal, we employed two differ- ent methodological approaches in primary-cultured rat hepatocytes (PCH) treated with E17G. First, we evaluated the ability of the NOX inhibitor apocynin (Apo) to prevent ERK 1/2 activation by measuring by western blotting (WB) its degree of phosphorylation. Additionally, we tested the possibility that ERK 1/2 inhibition with the MEK inhib- itor PD98059 (PD) can avoid the ROS increase induced by E17G, assessed fluorometrically by the 2’,7’-dichlorofluorescin-diacetate assay, which correlates dichlorofluorescin (DCF) fluorescence in- tensity with intracellular ROS concentration. E17G (200 μM, 20 min) increased by 54±14% (154±14, p