Mechanistic Insights Into Astragali Radix n‐Butanol on Podocyte Injury via Cell Cycle Regulation

The traditional Chinese medicine Astragali Radix (AR) shows therapeutic effects on nephrotic syndrome (NS), with AR n-butanol (AR-B) particularly ameliorating podocyte injury. Nevertheless, the molecular mechanisms are unclear. This study investigates how serum containing AR-B (AR-B-S) mitigates podocyte injury. Using rats as serum donors, we established an adriamycin (ADR)-induced injury model. The findings indicated that the optimal preparation protocol for AR-B-S was as follows: using serum collected 3.5 h after administration, incubating it at 56°C for 30 min, and employing a final concentration of 2.5%. Subsequently, compositional analysis using UPLC-MS technology and integrated network pharmacology predicted potential pathways, with more than 30% of AR-B-S targets for amelioration of podocyte injury related to the cell cycle. Furthermore, ADR was shown to facilitate the transition of MPC5 cells from a quiescent to a differentiated state. Additionally, ADR was demonstrated to have the ability to cause mitotic arrest in podocytes, which in turn leads to apoptosis. Conversely, intervention with AR-B-S was found to reverse ADR-induced mitotic arrest through the PI3K/Akt pathway, reducing podocyte apoptosis. This study showed AR-B-S reduces podocyte damage by regulating the cell cycle, providing a methodological reference for herbal serum preparation and suggesting a new therapeutic target for kidney disease.