The invention discloses application of schisandrol B in preparation of a medicine for preventing and treating cholestatic liver disease. By giving an excess amount of lithocholic acid, a mice intrahepatic cholestasis model is made, or a mice extrahepatic cholestasis model is made by bile duct ligation, then the mice intrahepatic cholestasis model and the mice extrahepatic cholestasis model are respectively collaboratively given the schisandrol, liver injury can be effectively alleviated, the bile acid level is reduced, glutamic-pyruvic transaminase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activity are decreased significantly, liver tissue necrosis degree is significantly alleviated, at the same time, total bile acid and total bilirubin content in serum are significantly reduced, and the total bile acid and total bilirubin content discharged with urine are increased. The results show that the schisandrol B can prevent and treat cholestatic liver injury induced by lithocholic acid and bile duct ligation, and prove that the effect mechanism may be involved in regulation of CYP3A11, UGT1A1 and OATP1A4 mediated bile acid metabolism and excretion.