Objective To investigate the mechanism of the action of nuclear factor erythroid 2-related factor 2Nrf2/solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) signaling pathway in NAFLD by in vivo and in vitro experiments. The mechanism of Gegen QinLian Decoction in the treatment of non-alcoholic fatty liver disease (NAFLD) was investigated in vivo and in vitro. Methods Animal experiments: Rats were fed with high-fat chow for 24 weeks to complete the induction modeling of NAFLD, and were randomly divided into normal, model, Gegen QinLian Decoction high-dose, medium-dose, low-dose, and metformin (Met) groups. From the 25th week onwards, the corresponding drugs were gavaged for two weeks according to the grouping until sampling. Biochemical kits were used to measure the levels of oxidative stress: malondialdehyde (MDA) and glutathione (GSH) in the liver tissues of rats in each group. A ferrous ion kit was used to detect ferrous iron (Fe2+) in rat liver tissues. qRT-PCR was used to detect Nrf2, heme oxygenase-1 (HO-1), SLC7A11, glutathione synthetase (GSS), GPX4 and acyl coenzyme A synthetase 4 in rat liver tissues. Glutathione Synthetase (GSS), GPX4 and Acyl Coenzyme A synthase (ACSL4) mRNA expression levels. Cellular experiments: 1 mmol/L free fatty acid (FFA) was used to induce lipid accumulation in hepatocellular carcinoma HepG2 cells mimicking the NAFLD in vitro model, and different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment. Oil red O staining was used to detect the lipid accumulation of cells in each group. Kits were used to determine the MDA and GSH content of HepG2 cells in different groups. Cell-specific ferrous kit was used to detect the ferrous content of cells in each group. qRT-PCR was used to detect the expression levels of Nrf2, HO-1, SLC7A11 , GSS, GPX4 and ACSL4 mRNA in cells of each group. Results Animal experiments: MDA and Fe2+ contents in liver tissues of rats in the model group were significantly higher than those in the blank group, and GSH contents were significantly lower (P<0.01); the Gegen QinLian Decoction high and medium dose groups and the metformin group could substantially reduce MDA and Fe2+ contents and elevate GSH contents, compared with the model group (P<0.01, 0.05). Compared with the model group, the Gegen QinLian Decoction high- and medium-dose groups and the metformin group increased the expression levels of Nrf2, HO-1, GSS, and GPX4 mRNA, and decreased the level of ACSL4 mRNA (P<0.01, 0.05). Cellular assay:Red lipid droplets were significantly increased in the HepG2 cell model group compared with the blank group, and the use of Gegen QinLian Decoction and metformin led to a significant decrease in lipid droplets. The kit assay revealed that MDA and Fe2+ content increased significantly in the HepG2 cell model group, and GSH content decreased significantly compared with the normal group (P<0.01),while all dose groups of Gegen QinLian Decoction and metformin group significantly decreased MDA and Fe2+ content compared with the model group (P<0.01), and the high and middle dose groups of Gegen QinLian Decoction, the low dose group, and the metformin group increased GSH content (P<0.01, 0.05). qRT-PCR experiments showed that the mRNAs of Nrf2, GSS, GPX4, and SLC7A11 in the high dose group of Gegen QinLian Decoction, Nrf2, HO-1, and SLC7A11 in the middle dose group, HO-1, SLC7A11, and GSS in the low dose group, as well as GSS, Nrf2, HO-1 in the metformin group, mRNA expression levels of GPX4 and SLC7A11 were significantly increased compared with the model group (P<0.01, P<0.05). The mRNA expression of ACSL4 was significantly decreased in the middle and low dose groups of Gegen QinLian Decoction and the metformin group (P<0.01, 0.05). Conclusion Gegen QinLian Decoction can improve NAFLD by inhibiting iron death, and its mechanism of action may be related to the regulation of Nrf2/SLC7A11/GPX4 signaling pathway.