We tested the hypothesis that differential expression of liver plasma membrane transporters might account for variations in biliary lipid secretion rates between gallstone-susceptible C57L/J and gallstone-resistant AKR/J mice. Plasma membrane fractions and total RNA isolated from livers of mice fed with a control or lithogenic (15% fat/1.25% cholesterol/0.5% cholic acid) diet were used for measurements of steady-state gene expression of hepatobiliary transport systems for bile salts (Ntcp1/Slc10a1, Oatp1/Slc21a1 and Bsep/Abcb11), phospholipids (Mdr2/Abcb4), organic anions (Mrp2/Abcc2) and organic cations (Oct1/Slc22a1). Irrespective of the diet, the steady-state gene expression of hepatobiliary transporters did not differ significantly between the two strains. Despite a higher basal bile flow and bile-salt secretion in C57L mice, Mrp2 (Abcc2) and Bsep (Abcb11) expression did not differ between the two strains. Elevated biliary phospholipid secretion in response to the lithogenic diet was linked to increased Mdr2 (Abcb4) protein expression, whereas the induction of Oct1 (Slc22a1) might reflect an enhanced uptake of choline for augmented phospholipid synthesis. In response to the lithogenic diet, Bsep (Abcb11) protein expression was up-regulated only marginally and bile salt secretion did not increase. The down-regulation of Ntcp1 (Slc10a1) protein expression might protect hepatocytes from high intracellular bile-salt loads. We conclude that variations in protein function rather than in the gene expression of liver plasma membrane transporters might account for variations in biliary lipid secretion rates. Our findings support the concept that the formation of lithogenic bile is caused by the hypersecretion of bile salts as a result of augmented availability of canalicular membrane cholesterol, possibly amplified by bile-salt—phospholipid uncoupling due to the increased bile flow.
- biliary lipids
- Lith genes
- organic anions
- organic cations
↵1 Present address: Department of Gastroenterology, Endocrinology and Hepatology, Robert-Bosch-Hospital, Auerbach-Strasse 110, D-70376 Stuttgart, Germany.
Abbreviations used: Bsep (Abcb11), bile-salt export pump; DPP IV, dipeptidyl peptidase IV; Gapdh, gene encoding glyceraldehyde-3-phosphate dehydrogenase; Mdr2 (Abcb4), multidrug resistance 2 P-glycoprotein; Mrp2 (Abcc2), multidrug-resistance-associated protein 2; Ntcp1 (Slc10a1), Na+-dependent taurocholate co-transporting protein 1; Oatp1 (Slc21a1), organic-anion-transporting polypeptide 1; Oct1 (Slc22a1), organic-cation-transporting protein 1; RT—PCR, reverse-transcriptase-mediated PCR.
- The Biochemical Society, London ©2002