NADPH OXIDASE PARTICIPATE IN THE CHOLESTATIC ALTERATIONS INDUCED BY ESTRADIOL 17 ΒETA-D-GLUCURONIDE DOWNSTREAM MEK-ERK 1/2 KINASES

SALAS GIMENA; ANDERMATTEN ROMINA BELEN; MEDEOT ANABELA CAROLINA; SCHUCK VIRGINIA SOLEDAD; RAZORI VALERIA; CIRIACI NADIA; BASIGLIO CECILIA L; CROCENZI FERNANDO ARIEL; MISZCUK GISEL SABRINA

Mar del plata

Congreso; Reunión Conjunta SAIC.SAI.AAFE.NANOMED.AR; 2021

SAIC.SAI.AAFE.NANOMED.AR

We have previously demonstrated that NADPH oxidase (NOX)-generated reactive oxygen species (ROS) are involved in the impairment of canalicular secretion induced by estradiol 17β-D-glucuronide (E17G) [Physiological Mini Reviews 12 (Special Edition): 019, 2019], and that NOX is in the same pathway that MEK-ERK ½ [Medicina (Bs. As.), 80 (supl V): 67, 2020]. Now, we intended to locate NOX into this pathway. To achieve this goal, we employed two different 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 inhibitor PD98059 (PD) can avoid the ROS increase induced by E17G, assessed fluorometrically by the 2?,7?-dichlorofluorescin-diacetate assay, which correlates dichlorofluorescin (DCF) fluorescence intensity with intracellular ROS concentration. E17G (200 µM, 20 min) increased by 54±14% (154±14, p