Astragalus root, a traditional Chinese herbal remedy, has shown potential benefits against diabetic nephropathy (DN). However, the mechanisms driving its effects remain poorly understood. This study explored the molecular pathways through which Astragalus root improves DN. To identify possible targets and mechanisms of Astragalus root in DN treatment, we applied network pharmacology, molecular docking, molecular dynamics simulation, and in vitro assays. Network pharmacology screening uncovered 46 overlapping targets between Astragalus root and DN. Protein–protein interaction (PPI) network analysis identified five core candidate targets: CASP3, VEGFA, CTNNB1, MYC, and PRKCB (PKCβ). KEGG pathway analysis indicated that the AGE-RAGE signaling pathway was the most significantly enriched. Molecular docking revealed that quercetin, β-carotene, daidzein, capsaicin, and kaempferol—potential bioactive components of Astragalus root—bound strongly to each of the five core targets. Molecular dynamics simulations further confirmed the conformational stability of kaempferol when complexed with these target proteins. In vitro experiments showed that kaempferol markedly reduced protein levels of α-SMA, Col I, and Col IV; lowered secretion of TNF-α, IL-6, and IL-1β; and decreased ROS and MDA content. Additionally, kaempferol’s therapeutic effects were mediated through suppression of the AGE-RAGE-PKCβ-TGF-β1 signaling axis. This work identified kaempferol, a bioactive ingredient of Astragalus root, as a potential therapeutic agent against DN, along with its target pathways. These findings provide a scientific foundation for its clinical translation.

Background/Objectives: Astragalus root, a traditional Chinese herbal remedy, has shown potential benefits against diabetic nephropathy (DN). However, the mechanisms driving its effects remain poorly understood. This study explored the molecular pathways through which Astragalus root improves DN. Methods: To identify possible targets and mechanisms of Astragalus root in DN treatment, we applied network pharmacology, molecular docking, molecular dynamics simulation, and in vitro assays. Results: Network pharmacology screening uncovered 46 overlapping targets between Astragalus root and DN. Protein-protein interaction (PPI) network analysis identified five core candidate targets: CASP3, VEGFA, CTNNB1, MYC, and PRKCB. KEGG pathway analysis indicated that the AGE-RAGE signaling pathway was the most significantly enriched. Molecular docking revealed that quercetin, β-carotene, daidzein, capsaicin, and kaempferol—major bioactive components of Astragalus root—bound strongly to each of the five core targets. Molecular dynamics simulations further confirmed the conformational stability of kaempferol when complexed with these target proteins. In vitro experiments showed that kaempferol markedly reduced protein levels of α-SMA, Col I, and Col IV; lowered secretion of TNF-α, IL-6, and IL-1β; and decreased ROS and MDA content. Additionally, kaempferol's therapeutic effects were mediated through suppression of the AGE-RAGE-PKC-TGF-β signaling axis. Conclusions: This work identified kaempferol, a bioactive ingredient of Astragalus root, as a potential therapeutic agent against DN, along with its target pathways. These findings provide a scientific foundation for its clinical translation.