Objective  To investigate the mechanism of AMP-activated protein kinase (AMPK)-mammalian target of rapamycin (mTOR) signaling pathway involved in acute kidney injury in mice. Methods  Thirty cases of healthy male C57BL/6 mice were divided into three groups-control group, model group and SB2111 group, with 10 mice in each group. Both the model group and the SB2111 group were treated with lipopolysaccharide (LPS) to establish acute kidney injury model, the control group were replaced with equal dose of normal saline, the SB2111 group were treated with 2 mL of AMPK blocker SB2111. The general condition and pathological characteristics of the mice were observed and recorded, and renal function were detected. Results   The body weight of the model group and the SB2111 group were significantly lower than those of the control group (P<0.05), and the kidney of the model group were significantly higher than the control group (P<0.05) that there were no significant difference compared between the SB2111 group and the control group (P>0.05). After the experiment, the 24 h urine protein, serum creatinine and urea nitrogen in the model group and SB2111 group were higher than the control group, and the SB2111 group were lower than the model group that compared the difference were statistically significant (P<0.05). The renal tubular epithelial cells in the control group were rich in cytoplasm, the glomeruli and renal tubules in the renal tissues were normal, the glomerular congestion and tubular formation in the model group, the glomerular congestion in the SB2111 group, and the proximal koji, tubular epithelial cells were slightly swollen with small amount of inflammatory cell infiltration. The relative expression levels of TNF-α, TGF-β1, AMPK and mTOR in the model group and SB2111 group were significantly higher than those in the control group, and the SB2111 group were lower than the model group that compared the difference were statistically significant (P<0.05). Conclusions   AMPK-mTOR signaling pathway are activated in acute kidney injury in mice. Inhibition of AMPK-mTOR signaling pathway can inhibit the expression of TNF-α and TGF-β1 and promote the recovery of renal function in mice.