Objective To investigate the effect of rapamycin on primary sclerosing cholangitis in C57BL/ 6J mice induced by 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) and to explore its potential mechanism. Methods 6 ~ 8- week-old female C57BL/ 6J mice were randomly divided into a normal diet group ( healthy control group), a 0. 1% DDC diet model group (DDC group), and a DDC with rapamycin treatment group (RAPA group). Rapamycin (5 mg / kg body weight) was injected intraperitoneally every day for 1 week. Hematoxylin-eosin and Masson staining were used to semiquantitatively evaluate the degree of pathological changes and liver fibrosis in liver tissues, respectively. Immunohistochemistry was used to detect CK-19 and Ki-67 and to quantitatively analyze the degree of biliary duct proliferation in liver tissues. Real-time quantitative polymerase chain reaction was used to detect the expression of inflammatory cytokines (IL-6, TNF-α, IL-1β) and fibrosis-related molecules ( TIMP, TGF-β, and collagen I) at the mRNA level. Western blot was used to detect the expression of Akt / mTOR/ NF-κB signaling pathway proteins and their phosphorylation levels. Results Compared with the DDC group, the RAPA group showed a marked reduction in hepatic inflammation and biliary duct hyperplasia as well as amelioration of hepatic fibrosis. Inflammatory cytokines (IL-6, TNF-α, IL-1β) and fibrosis-related molecules ( TIMP, TGF-β, and collagen I) were significantly decreased; furthermore, the phosphorylation levels of Akt / mTOR/ NF-κB signaling pathway proteins were also remarkably decreased. After rapamycin treatment, bile duct injuries in the DDC group as indicated by expression of CK-19 and Ki-67 were also significantly improved. Conclusions Rapamycin can alleviate DDC-induced primary sclerosing cholangitis in mice by inhibiting the Akt / mTOR/ NF-κB signaling pathway.