Functional processing enhances hepatic targeting: the OAT2/MRP2 mechanism of vinegar-processed Cyperi Rhizoma

Processing techniques play a critical role in modifying the bioavailability of botanical ingredients. Traditional processing theory suggests that vinegar processing enhances the hepatic-targeting effects of Cyperi Rhizoma (CR), a botanical used in functional foods and traditional medicine. However, the molecular mechanism remains largely unexplored. This study integrated in silico docking with in vitro experiments to investigate vinegar processing on hepatic uptake of CR bioactive components. HS-GC–MS confirmed a key chemical transformation: conversion of cyperene to cyperotundone in vinegar-processed CR (VCR). Molecular docking predicted high affinity of VCR-derived components, particularly cyperotundone, for hepatic transporters. In HepaRG cells, UPLC-QqQ-MS demonstrated significantly higher intracellular accumulation of bioactive components from VCR versus raw CR (RCR). Ligand fishing assays validated strong binding between VCR components and hepatic uptake transporters. Mechanistically, VCR and cyperotundone upregulated the uptake transporter OAT2 while inhibiting the efflux transporter MRP2. Transporter overexpression confirmed that OAT2 facilitates, while MRP2 limits, cellular uptake. In conclusion, vinegar processing enhances hepatic retention of CR active components by modulating OAT2 and MRP2. These integrated findings provide a scientific basis for vinegar-processed CR as a liver-protective functional food ingredient.