Cav‐1 regulates the bile salt export pump on the canalicular membrane of hepatocytes by PKCα‐associated signalling under cholesterol stimulation

Abstract

AbstractBackground and AimsThe secretion of bile salts transported by the bile salt export pump (BSEP) is the primary driving force for the generation of bile flow; thus, it is closely related to the formation of cholesterol stones. Caveolin‐1 (Cav‐1), an essential player in cell signalling and endocytosis, is known to co‐localize with cholesterol‐rich membrane domains. This study illustrates the role of Cav‐1 and BSEP in cholesterol stone formation.MethodsAdult male C57BL/6 mice were used as an animal model. HepG2 cells were cultured under different cholesterol concentrations and BSEP, Cav‐1, p‐PKCα and Hax‐1 expression levels were determined via Western blotting. Expression levels of BSEP and Cav‐1 mRNA were detected using real‐time PCR. Immunofluorescence and immunoprecipitation assays were performed to study BSEP and Hax‐1 distribution. Finally, an ATPase activity assay was performed to detect BSEP transport activity under different cholesterol concentrations in cells.ResultsUnder low‐concentration stimulation with cholesterol, Cav‐1 and BSEP protein and mRNA expression levels significantly increased, PKCα phosphorylation significantly decreased, BSEP binding capacity to Hax‐1 weakened, and BSEP function increased. Under high‐concentration stimulation with cholesterol, Cav‐1 and BSEP protein and mRNA expression levels decreased, PKCα phosphorylation increased, BSEP binding capacity to Hax‐1 rose, and BSEP function decreased.ConclusionCav‐1 regulates the bile salt export pump on the canalicular membrane of hepatocytes via PKCα‐associated signalling under cholesterol stimulation.