Triptolide (TP) can cause severe hepatotoxicity which limits its clinical application.Isoliquitigenin (ISL) has been reported to protect against TP-induced liver injury, but the mechanisms are not fully elucidated. This study aims to explore the role of nuclear transcription factor E2-related factor 2 (Nrf2) and hepatic transporters in TP-induced hepatotoxicity and the reversal effects of ISL on it. TP treatment caused cytotoxicity in human L02 hepatocytes and acute liver injury in ICR mice reflected by changes in liver indices, histopathology, serum biochemical indices, as well as the hepatic bile acid profiles; while combined treatment with ISL effectively alleviated TP-induced hepatotoxicity.Further, compared with the TP-injured group, ISL pretreatment enhanced Nrf2 expressions and nuclear accumulations as well as expressions of its downstream NAD(P)H:quinine oxidoreductase 1 (NQO1); expressions of hepatic transporters P-glycoprotein (P-gp), multidrug resistance proteins (MRPs), bile salt export pump (Bsep), and organic anion transporting polypeptide (Oatp) were also induced. In addition, vesicular transport assays identified that neither was TP exported by MRP2, nor did TP influence the transport activities of P-gp and MRP2; and uptake assays showed that TP was also not a substrate of both OATP1B1 and OATP1B3. Overall, these results indicate that ISL may reduce the hepatic oxidative stress and accumulations of both endogenous bile acids and exogenous TP and its metabolites by enhancing expressions of Nrf2 and NQO1 as ell as hepatic influx and efflux transporters. Since TP does not influence the transport activities of P-gp and MRP2 and is not transported by MRP2, OATP1B1, and OATP1B3, effects of TP on hepatic transporters are mainly at transcriptional levels, and changes of hepatic bile acid profiles may be of great significance in the mechanisms of TP-induced hepatotoxicity.