To investigate the chemical composition of the aqueous extract of Cistanche tubulosa(AECT) and the mechanism of action of AECT in the treatment of diabetic nephropathy (DN),UPLC-MS/MS was used to analyse and identify the compounds,Using PubChem,GeneCards,OMIM,DAVID and other databases,we predicted the targets and pathways of the compounds to prevent and control DN,performed GO and KEGG enrichment analysis on the core genes,and constructed a visual ‘compound -target - pathway’ network diagram using Cytoscape software.Determination of Anti-DN Activity of AECT by MTT Method,Biochemical kits were used to detect the activity of superoxide dismutase (SOD) and the levels of glutathione peroxidase (GSH-Px).ELISA was used to measure the levels of inflammatory factors interleukin-1β (IL-1β),tumour necrosis factor (TNF-α),and transforming growth factor (TGF-β).Detection of apoptosis was performed using the Annexin V-FITC/PI assay.Western blot was performed to detect the expression of relevant proteins.AECT contained 84 chemical components,with 15 compounds identified as possible cleavage pathways.Screening 70 potential targets and multiple signaling pathways of AECT for the treatment of DN.The results of the in vitro experiments showed that AECT increased the survival of HK-2 cells and inhibited apoptosis in a high-sugar and high-fat environment,And SOD activity and GSH-Px content were up-regulated,while the expression levels of inflammatory factors IL-1β,TNF-α and TGF-β were inhibited.The Western blot results indicate that AECT has an impact on the PI3K-AKT signaling pathway.This study investigates the potential targets and mechanisms of action of the main active ingredients in AECT for the treatment of DN.The findings suggest avenues for further research into the use of AECT for DN treatment.