A top-down approach for integrative analysis of vinegar processing effects on Cyperi Rhizoma

Cyperi Rhizoma (CR) is a commonly used medicinal plant for hepatic protection in clinical practice. Vinegar processing, a vital and indispensable step for CR, is regarded as an amplifier that improves the efficacy of CR in hepatic diseases. This study aimed to apply a top-down approach to explore the effect of vinegar processing on CR. After integrating chemical and bioactive information, eleven chemical compounds (including tryptophan, chlorogenic acid, catechin, epicatechin, protocatechuic aldehyde, protocatechuic acid, cyperenone, α-cyperone, 4-coumaric acid, ferulic acid and luteolin) are identified as vinegar-processing chemical markers (VP markers) of CR based on UHPLC-Q-TOF-MS combing with UHPLC-QQQ-dMRM-MS/MS. The identified VP markers possess high correlation with the increased antioxidant activity induced by vinegar processing. Furthermore, vinegar processing optimized by Box-Behnken design-response surface methodology (BBD-RSM) significantly enhances the anti-hepatic fibrosis efficacy of CR, reversing CCl₄-induced abnormalities in liver function (ALT, AST, LDH), fibrotic progression (α-SMA, Tgfb1, Col1a1), and plasma metabolites (lysoPCs, PCs, steroids, etc) in rats. This study is meaningful for exploring the therapeutic substances of CR and promoting its further development.