Spironolactone ameliorates lipopolysaccharide-induced cholestasis in rats by improving Mrp2 function: Role of transcriptional and post-transcriptional mechanisms

MARTÍN, PAMELA L.; CIRIACI, NADIA; BASIGLIO, CECILIA L.; RAZORI, MARÍA VALERIA; BAROSSO, ISMAEL R.; SÁNCHEZ POZZI, ENRIQUE J.; ROMA, MARCELO G.; RAZORI, MARÍA VALERIA; BAROSSO, ISMAEL R.; SÁNCHEZ POZZI, ENRIQUE J.; ROMA, MARCELO G.; MAIDAGAN, PAULA M.; ANDERMATTEN, ROMINA B.; RUIZ, MARÍA LAURA; MAIDAGAN, PAULA M.; ANDERMATTEN, ROMINA B.; RUIZ, MARÍA LAURA; MARTÍN, PAMELA L.; CIRIACI, NADIA; BASIGLIO, CECILIA L.

LIFE SCIENCES

PERGAMON-ELSEVIER SCIENCE LTD

Lugar: Amsterdam; Año: 2020 vol. 259 p. 118352 - 118352

0024-3205

Aims: Lipopolysaccharide (LPS) induces inflammatory cholestasis by impairing expression, localization, and function of carriers involved in bile formation, e.g. bile salt export pump (Bsep) and multidrug resistance-associated protein 2 (Mrp2). A specific therapy against this disease is still lacking. Therefore, we evaluated the anticholestatic effects of spironolactone (SL), a PXR ligand that regulates bile salt homeostasis, up-regulates Mrp2, and bears anti-inflammatory properties. Main methods: Male Wistar rats were divided into four groups: Control, SL (83.3 mg/kg/day of SL, i.p., for 3 days), LPS (2.5 mg/kg/day, i.p., at 8 am of the last 2 days, and 1.5 mg/kg/day at 8 pm of the last day), and SL + LPS. Biliary and plasma parameters and the expression, function, and localization of Mrp2 and Bsep were evaluated. Key findings: SL partially prevented LPS-induced drop of basal bile flow by normalizing the bile salt-independent fraction of bile flow (BSIBF), via improvement of glutathione output. This was due to a recovery in Mrp2 transport function, the major canalicular glutathione transporter, estimated by monitoring the output of its exogenously administered substrate dibromosulfophthalein. SL counteracted the LPS-induced downregulation of Mrp2, but not that of Bsep, at both mRNA and protein levels. LPS induced endocytic internalization of both transporters, visualized by immunofluorescence followed by confocal microscopy, and SL partially prevented this relocalization. SL did not prevent the increase in IL-1β, IL-6, and TNF-α plasma levels. Significance: SL prevents the impairment in Mrp2 expression and localization, and the resulting recovery of Mrp2 function normalizes the BSIBF by improving glutathione excretion.