Hepatoprotective and Antioxidant Activities of a Medicine-Food Homology Formula and Its <i>Lactiplantibacillus plantarum</i>-Fermented Product Against Acetaminophen-Induced Oxidative Stress in HepG2 Cells

Abstract

Background: Acetaminophen (APAP) overdose depletes glutathione (GSH) and induces oxidative liver injury. The medicine-food homology formula (MFHF) is composed of Chrysanthemi flos (CF), Cassiae semen (CS), and Lycii fructus (LF) (3:20:10, w/w/w). However, the effects of MFHF and its Lactiplantibacillus plantarum-fermented products (MFHFF) on quality markers and activities remain unclear. Objectives: This study investigated the prebiotic potential of MFHF; examined the stability of chlorogenic acid and chrysophanol before and after fermentation; compared hepatoprotective effects between MFHF and MFHFF in APAP-treated HepG2 cells; evaluated antioxidant activities; and assessed intra-formula interactions (additive/synergistic) among CF, CS, and LF. Methods: Fermentations were performed with L. plantarum BCRC12251 in 0 - 5% (w/v) MFHF substrates. Viable counts and pH were used to assess prebiotic effects. The HPLC was used to determine the content of chlorogenic acid and chrysophanol before and after fermentation. Antioxidant activities were measured by DPPH and potassium ferricyanide reducing antioxidant power (PFRAP) assays. Hepatoprotection was evaluated in APAP-treated HepG2 cells using cell viability, intracellular GSH, and malondialdehyde (MDA) as endpoints. Intra-formula interactions were evaluated by comparing observed and expected values. Data were analyzed using independent-sample t-tests and one-way ANOVA followed by Duncan’s multiple range test (P < 0.05). Results: MFHF enhanced L. plantarum growth by 26-fold and retained phytochemical markers after fermentation. Both MFHF and MFHFF increased viability and GSH and decreased MDA in a dose-dependent manner in APAP-challenged HepG2 cells. MFHF and MFHFF showed comparable antioxidant activities. Viability and DPPH activity were synergistic, whereas GSH, MDA, and PFRAP showed additive effects. Conclusions: MFHF could act as a prebiotic for L. plantarum. Fermentation preserved MFHF’s chemical integrity and hepatoprotective efficacy. Both preparations demonstrated comparable antioxidant and cytoprotective properties, supporting their potential as multifunctional candidates against oxidative liver injury in vitro and warranting further in vivo validation.