Cholestasis is associated with retention of bile acids and reduced expression of the Na⁺/ taurocholate cotransporter (Ntcp), the major hepatocellular bile acid uptake system. This study aimed to determine whether (i) down-regulation of Ntcp in obstructive cholestasis is a consequence of bile acid retention and (ii) is mediated by induction of the transcriptional repressor short heterodimer partner 1 (SHP-1). To study the time course for the changes in serum bile acid levels as well as SHP-1 and Ntcp steady-state mRNA levels, mice were subjected to common bile duct ligation (CBDL) for 3, 6, 12, 24, 72 and 168 hours (h) and compared to sham-operated controls. Serum bile acid levels were determined by radioimmunoassay. SHP-1 and Ntcp steady-state mRNA expression were assessed by Northern blotting. In addition, Ntcp protein expression was studied by Western blotting and immunofluorescence microscopy. Increased SHP-1 mRNA expression paralleled elevations of serum bile acid levels and was followed by down-regulation of Ntcp mRNA and protein expression in CBDL mice. Maximal SHP-1 mRNA expression reached a plateau phase after 6 h CBDL (12-fold; p<0.001) and preceded the nadir of Ntcp mRNA levels (12%, p<0.001) by 6 h. In conclusion, bile acid-induced expression of SHP-1 may at least in part mediate down-regulation of Ntcp in CBDL mice. These findings support the concept, that down-regulation of Ntcp in cholestasis limits intracytoplasmatic accumulation of potentially toxic bile acids.