Objective To investigate the effect and mechanism of andrographolide (AG) on lipopolysaccharide (LPS)-induced ferroptosis in renal tubular epithelial cells (HK-2 cells). Methods HK-2 cells were treated with LPS to simulate the in vitro HK-2 injury model of sepsis. The cells were further treated with AG of 5, 10, 20, 40 μmol/L and randomly divided into control group, LPS group, LPS+dimethyl sulfoxide group (DMSO group), and AG group. Cell viability was detected by the CCK-8 method, and the optimal concentrations of LPS and AG were screened. Cell morphological change, the levels of kidney injury markers, including neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), malondialdehyde (MDA), glutathione (GSH) and reactive oxygen species (ROS), as well as the expression levels of ferroptosis regulatory proteins such as solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4) and ferritin in each group were compared, and the protective effect of AG treatment on the cells was evaluated. Results Compared with the control group, the cell viabi-lity and GSH content decreased significantly in HK-2 cells treated with 10 μg/mL LPS; cell shrinkage and adhesion ability were poor; the contents of oxidative products MDA and ROS, as well as the levels of kidney injury markers NGAL and KIM-1 increased significantly, while expression levels of SLC7A11 and GPX4 protein decreased; ferritin expression level increased; differences were all statistically significant (all P < 0.05). Compared with LPS group, the cell viability, GSH content, as well as protein expression levels of SLC7A11 and GPX4 increased significantly after AG intervention, while ferritin expression level decreased, differences were all significant (all P < 0.05). MDA content, ROS fluorescence intensity, and the levels of kidney injury markers NGAL and KIM-1 decreased significantly, difference were all significant (all P < 0.05). Conclusion AG has a protective effect on LPS-induced HK-2 cell injury, possibly by activating SLC7A11/GPX4 pathway, reducing oxidative stress, up-regulating antioxidant enzyme activity, and alleviating ferroptosis.