〔Abstract〕 To investigate the role of transcription factor EB (TFEB) -autophagy pathway in rifampicin- induced liver injury and its possible mechanism . Methods Forty 6-8-week-old C57/BL6 mice were randomly di- vided into five groups : control group , model group , TFEB low-dose agonist group , TFEB high-dose agonist group , and autophagy agonist group , with 8 mice in each group . Except for the control group , the other four groups were given rifampicin 200 mg/(kg ·d) by gavage daily . TFEB agonist was administered intraperitoneally at a low dose of 20 mg/kg and a high dose of 50 mg/kg for 7 days at 1 h after rifampicin administration . Autophagy agonist was administered by gavage at a dose of 10 mg/kg 6 h before rifampicin administration on day 1 . The experiment was completed 7 days after modeling. The degree of liver injury was evaluated by detecting liver function indexes and liver pathological changes . Western blot was used to detect the protein expression total TFEB , chelator 1( p62) , microtubule-associated protein light chain 3(LC3) , benzyl chloride 1(Beclin-1) , sodium taurocholate co-transpor- ting polypeptide(NTCP) and bile salt export pump(BSEP) levels in liver nucleus/liver tissue were quantified . Re- sults Compared with the control group , the serum levels of alanine aminotransferase ( ALT) , aspartate amin- otransferase (AST) , total bilirubin ( TBIL) , direct bilirubin ( DBIL) , and total bile acid ( TBA) in the model group increased (P < 0. 05) , and obvious pathological changes were observed in the liver. Compared with the mod- el group , the high dose and low dose of TFEB agonist and autophagy agonist groups had reductions in the above in- dicators (P < 0. 05) . Compared with the low-dose TFEB agonist group , the high-dose TFEB agonist group had re- ductions in the above indicators (P < 0. 05) . The proportion of TFEB in the nucleus was ( 1 . 0 ±0. 10) in the con- trol group , (0. 6 ±0. 05) in the model group , (0. 8 ±0. 08) in the low-dose agonist group , and (0. 9 ±0. 07) in the high-dose agonist group (P < 0. 05) . Autophagy agonist group (0. 7 ±0. 06) (P < 0. 05) . Compared with the control group , the levels of NTCP and BSEP in the liver of the model group decreased (P < 0. 05) , and the expres- sion of NTCP and BSEP in the TFEB low-dose and high-dose agonist groups were restored , and the expression of NTCP and BSEP in the autophagy agonist group also increased (P < 0. 05) . Compared with the control group , the protein expression levels of TFEB , LC3-Ⅱ/LC3 - Ⅰand Beclin-1 in the liver tissue of the model group significantly decreased (P < 0. 05) , while the protein expression level of p62 significantly increased ( P < 0. 05) . Compared with the model group , the protein expression levels of TFEB , LC3-Ⅱ/LC3 - Ⅰand Beclin-1 in the liver tissue of the TFEB agonist high-dose group , low-dose group and autophagy agonist group increased (P < 0. 05) , while the pro- tein expression level of p62 decreased (P < 0. 05) . Conclusion TFEB can improve rifampicin-induced liver inju- ry by activating autophagy pathway , and the main mechanism may be related to the up-regulation of NTCP and BSEP expression .