Paeoniflorin alleviates nephrolithiasis by targeting serpine1 and inhibiting the NF-κB signaling pathway.

Paeoniflorin (PF) is a bioactive monoterpene glycoside with well-established anti-inflammatory properties; however, its therapeutic potential and molecular mechanism in nephrolithiasis remain insufficiently defined. In this study, network pharmacology was employed to predict potential targets of PF. In vitro, HK-2 cells exposed to calcium oxalate (CaOx) crystals were treated with PF to evaluate inflammatory responses, crystal adhesion, and NF-κB activation. A glyoxylic acid-induced mouse model was used to assess renal crystal deposition, renal function, and expression of key regulatory targets, supported by transcriptomic profiling. Molecular docking and molecular dynamics simulations were performed to analyze the interaction between PF and Serpine1, and functional relevance was further examined using exogenous Serpine1 rescue experiments. Integrated analyses indicated that PF exerts anti-nephrolithic effects predominantly through modulation of inflammation-associated signaling. PF dose-dependently suppressed CaOx-induced NF-κB phosphorylation and nuclear translocation, reduced the release of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β), and decreased crystal adhesion in HK-2 cells. In vivo, PF markedly attenuated renal crystal deposition, improved renal functional indices, and downregulated renal Serpine1 and IL-1β expression. Computational simulations supported a stable interaction between PF and Serpine1 (binding energy − 6.22 kcal/mol). Consistently, supplementation with exogenous Serpine1 partially reversed PF-mediated inhibition of NF-κB activation and cytoprotective effects. Collectively, these findings suggest that PF mitigates CaOx-associated renal injury by suppressing inflammation-centered signaling programs involving the Serpine1/NF-κB axis, thereby providing mechanistic insight into PF as a potential multi-target therapeutic candidate for nephrolithiasis.