Objective The in vitro co-incubation method was employed to investigate the metabolic transformation regularity of Semen Cuscutae-Fructus Lycii (SC-FL) mediated by gut microbiota in premature ovarian insufficiency (POI) rats. Methods SC-FL and its thirteen monomer components were co-incubated with gut microbial communities from POI rats in vitro, followed by UPLC-Q-TOF-MS for qualitative analysis. Results A total of 85 compounds were characterized from the SC-FL co-incubated samples, including 61 prototypical components (e.g., rutin), 13 metabolites (e.g., methyl hyperoside), and 11 kinds of components that exist in the prototype at the same time and can be metabolized by other components, (e.g., isorhamnetin). Hyperoside was characterized by five metabolites, rutin by seven, quercitrin by six, astragalin by seven, isoquercitrin by five, quercetin by seven, isorhamnetin by four, kaempferol by five, apigenin by four, chlorogenic acid by nine, ferulic acid by three, scopoline by four, and scopoletin lactone by three. Conclusion The metabolic transformation reactions mediated by the intestinal microbiota in POI rats mainly include Phase I metabolic reactions such as hydrolysis, reduction-hydrogenation, and hydroxylation reactions, and Phase II metabolic reactions such as methylation reactions. Reverse traceability analysis of the metabolic conditions of each component revealed that isorhamnetin, quercetin, kaempferol and apigenin were mutually metabolically transformed in the intestinal flora of POI rats These findings provide important evidence for elucidating the pharmacodynamic material basis and mechanisms of SC-FL in POI, and lay a foundation for subsequent research on optimizing SC-FL efficacy through gut microbiota modulation.

Semen Cuscutae-Fructus Lycii herb pair  /  premature ovarian insufficiency  /  UPLC-Q-TOF-MS  /  gut microbiota  /  metabolic pathways  /  isorhamnetin  /  quercetin  /  kaempferol  /  apigenin