Potential targets and molecular mechanisms of D-pinitol against acute kidney injury based on network pharmacology and experimental validation

Abstract

Abstract Acute kidney injury (AKI) is a severe clinical syndrome, with ischemia/reperfusion (I/R) being one of its most common causes. Although D‑pinitol (DP), an inositol‑like bioactive molecule, is known to confer renal protection, its efficacy against I/R‑induced AKI remains unknown. A mouse kidney I/R model was employed to evaluate the renoprotective effect of DP. Relevant targets associated with DP and AKI were retrieved from publicly available databases. Subsequently, network pharmacology analysis was conducted to identify the potential targets and signaling pathways. Molecular docking was then performed to predict the binding affinity of DP to core targets identified. Furthermore, in vivo and in vitro experiments were performed to validate these findings. Systemic toxicity was assessed by serological and histopathological examinations. The results show that DP significantly attenuated I/R-induced kidney dysfunction and apoptosis. Network pharmacology analysis identified 108 overlapping targets, with AKT1, HSP90AA1, SRC, CASP3, and MMP9 identified as core targets. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed PI3K/AKT signaling pathway as the primary mechanism. Consistent with these predictions, DP enhanced PI3K and AKT phosphorylation in kidney tissue. Molecular docking indicated that DP exhibited the strongest binding affinity to SRC, suggesting it as a potential target. In a hypoxia/reoxygenation (H/R)-induced human renal proximal tubular epithelial (HK-2) cell model, DP significantly increased the phosphorylation of SRC, PI3K, and AKT, and these effects were abrogated by the SRC specific inhibitor PP2. Collectively, DP alleviated I/R‑induced injury and apoptosis, potentially through activation of the SRC/PI3K/AKT signaling pathway.