Objective To induce iron overload in the liver of rats by iron dextran,observe the effect and mechanism of Xiaoyao Powder on liver cell iron death,and explore the role and position of Xiaoyao Powder's liver soothing,blood nourishing,and spleen strengthening effects in the entire formula through the analysis of various functional formulas.Methods A rat liver iron death model in-duced by chronic iron overload was established by intraperitoneal injection of iron dextran for 7 consecutive weeks.After three weeks of modeling,drug intervention was initiated,using Xiaoyaosan Quanfang,Xiaoyaosan Qushugan Yao,Xiaoyaosan Jianpi Yao,and Xi-aoyaosan Quxueyuan Yao respectively.In addition,deferoxamine was used as a positive control drug.Detect the content of reduced gluta-thione(GSH),malondialdehyde(MDA),ferrous ions,and lipid peroxidation ROS.RT-PCR and Western blot were used to detect the expression of BMP6/HJV/SMAD4 and downstream hepcidin ferroportin axis genes or proteins in rat liver tissue.Results The liver Fe2+,MDA and ROS contents of the model group rats were significantly increased compared to the control group,while the GSH content was sig-nificantly decreased.The mRNA expression of glutathione peroxidase 4(Gpx4)and membrane iron transporter(Fpn1)was significantly decreased,while the mRNA expression of prostaglandin peroxidase 2(Ptgs2)and hemodulin,as well as BMP6,HJV,and SMAD4 pro-teins,were significantly increased.Compared with the model group,the entire formula of Xiaoyaosan,as well as the various functional de-coctions and positive control groups,can significantly reverse the above indicators.Compared with the whole formula of Xiaoyaosan,the efficacy of Xiaoyaosan in each disassembled formula group was significantly reduced in reversing the above indicators(except for the Xi-aoyaosan spleen tonifying drug,which had no significant difference in SMAD4 protein expression compared to the whole formula group).Conclusion Xiaoyaosan can inhibit iron-induced liver ferroptosis in rats,which may be related to the regulation of the BMP6/HJV/SMAD4 pathway and its downstream hepcidin ferroportin axis.All functional components of Xiaoyaosan have played a role in inhibiting liv-er cell ferroptosis.