The rapid isolation of target constituents from natural products poses a significant challenge and is a key focus in current research. The Hosta plantaginea flower (HPF), a traditional Chinese medicinal herb, is primarily used to treat inflammatory diseases, with kaempferol as one of its major bioactive constituents. In this study, macroporous adsorption resin was used to purify total flavonoids (TF) from the HPFs. The 50% ethanol–water elution fraction of the TF was then recrystallized to yield kaempferol with a purity of 99.44%. Network pharmacology analysis identified 61 potential kaempferol-inflammation targets, which were linked to the PI3K-Akt and TNF signaling pathways. Molecular docking and molecular dynamics simulations revealed the stability and binding of kaempferol to PI3K, Akt, and TNF-α proteins. The analysis metrics included binding ability, the root mean square deviation (RMSD), radius of gyration, free energy landscape, solvent-accessible surface area, hydrogen bond count, RMS fluctuation, free binding energy, amino acid residue free energy decomposition, and principal component analysis. The anti-inflammatory mechanism of kaempferol was further validated in an LPS-induced RAW264.7 cell model, where it was shown to inhibit the PI3K-Akt and TNF-α signaling pathways. This study provides new insights into the anti-inflammatory mechanism of kaempferol and presents novel strategies for the rapid isolation of target constituents from natural products.

Background:Disrupted bile acid homeostasis represents a central feature of cholestatic liver injury, for which effective therapeutic options remain limited. Saxifraga umbellulata Hook. f. et Thoms. (SU) has traditionally been used in Tibetan medicine to treat hepatobiliary disorders, yet the pharmacological basis of SU remains unclear. Therefore, this study aimed to evaluate the effects of SU on cholestatic liver injury and explore the regulatory mechanisms underlying these effects.Methods:A mouse model of cholestatic liver injury was established by administering α-naphthyl isothiocyanate (ANIT). The effects of SU were evaluated using serum biochemical measurements, liver histopathology, and targeted profiling of hepatic bile acids. Protein expression associated with farnesoid X receptor (FXR) signaling was assessed by Western blotting in liver and ileal tissues. Representative triterpenoids in the SU extract were further examined using molecular docking and HepG2 cell experiments.Results:SU administration reduced serum markers of hepatocellular injury and cholestasis and lessened hepatic necrosis and inflammatory cell infiltration in ANIT-treated mice. SU also shifted the altered hepatic bile acid profile toward a more balanced state, including partial correction of the disturbed proportions of primary and secondary bile acids. These metabolic changes were accompanied by reduced hepatic CYP7A1 expression and increased expression of BSEP and MRP2. In addition, SU increased ileal FXR and FGF15 expression and hepatic FGFR4 expression, a pattern consistent with coordinated regulation of the intestinal FXR–FGF15/hepatic FGFR4 axis. Meanwhile, molecular docking and HepG2 cell experiments suggested that cucurbitacin IIb and saxifragic acid may contribute to the observed FXR-related responses.Conclusions:SU mitigated ANIT-induced cholestatic liver injury and promoted partial recovery of hepatic bile acid homeostasis. The protective effects of SU were associated with coordinated changes in hepatic and intestinal FXR-related signaling and with reduced inflammatory injury. These results provide experimental evidence supporting the traditional use of SU for hepatobiliary disorders and highlight cucurbitacin IIb and saxifragic acid as potential bioactive triterpenoids.