Objective To observe the effects of different ligation sites and fasting method on a C57BL/ 6J mouse model of partial bile duct ligation (pBDL)-induced cholestasis, to establish a pBDL modeling method with a high modeling rate, typical symptoms, and good stability. Methods C57BL/ 6J mice were subjected to selective ligation of the left hepatic bile duct (L-pBDL) and left-to-median bile duct junction ligation (ML-pBDL) for modeling, and the effects of different pBDL ligation method on serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase (ALP), total bilirubin, total bile acid, and liver histopathology were observed. The effects of different fasting method on symptoms and liver injury in the ML-pBDL model were also observed after fasting for 12 and 16 h before surgery, and for 4 h after surgery. Results (1)The incidence of jaundice in the ML-pBDL group was 52. 94% and the survival rate within 3 weeks after surgery was 64. 71%, while the incidence of jaundice in the L-pBDL group was 11. 76% and the survival rate within 3 weeks after surgery was 82. 35%. Compared with those in the sham surgery group, serum liver function indicators were significantly increased in the L-pBDL and ML-pBDL groups (P<0. 01), and ALP activity was significantly higher in the ML-pBDL group than in the L-pBDL group (P<0. 05). Compared with mice in the L-pBDL group, mice in the MLpBDL group had more severe liver fibrosis at 3 weeks post-surgery (P<0. 01). (2)In addition, the incidence of jaundice in the 16 h fasting group was 93. 33% and the survival rate within 3 weeks after surgery was 73. 77%, while the incidence of jaundice in the 12 h fasting group was 42. 86% and the survival rate within 3 weeks after surgery was 71. 42%.Compared with those in the normal group, ALP activity, alanine aminotransferase / aspartate aminotransferase ratio, total bile acid level, and proportion of collagen fiber area were all significantly increased in the 16 h and 12 h fasting groups (P<0. 05). Although the observed indicators were higher in the 16 h fasting group compared with those in the 12 h fasting group, the difference was not significant (P>0. 05). Mice in the 12 h and 16 h fasting groups both showed significant bile duct hyperplasia and liver fibrosis (P<0. 01), with more severe liver fibrosis in the 16 h fasting group (P<0. 01). Conclusions Both L-pBDL and ML-pBDL ligation method can be used to establish a mouse model of cholestasis;however, symptoms in the L-pBDL model only exhibit transient damage characteristics, while the liver lesions in the MLpBDL model are typical and stable. Prolonging the preoperative fasting time can improve the modeling rate and stability of the ML-pBDL model and produce more-typical pathological symptoms.