ObjectiveTo investigate the protective effect of Rosa roxburghii fermentation broth against acetaminophen (APAP)-induced drug-induced liver injury (DILI) in mice and explore its potential mechanism based on nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway. Methods Taking the R. roxburghii juice as the control, the main active component contents and in vitro antioxidant capacity of R. roxburghii fermentation broth were analyzed. A total of 60 Kunming mice were randomly divided into control group, model group, N-acetylcysteine (NAC, 0.15 g/kg) group, and R. roxburghii fermentation broth low-, medium-, and high-dose (5, 10, 20 mL/kg) groups, with 10 mice in each group. All groups were consecutively administered for 7 d, 1 h after the last administration, except for the control group, all other groups received a single intraperitoneal injection of APAP (400 mg/kg) to establish the acute liver injury model. After 12 h of modeling, the mice were anesthetized and euthanized. The intact liver of mice was immediately removed and observed for any visible damage or lesions. The body weight and liver weight were measured and liver index was calculated. Activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β in serum, as well as activity of superoxide dismutase (SOD) and levels of glutathione (GSH), malondialdehyde (MDA) in liver tissue were detected by ELISA. Hepatic pathological changes were observed by hematoxylin-eosin (HE) staining. The mRNA and protein expressions of Nrf2, heme oxygenase-1 (HO-1), SOD-1 and catalase (CAT) in liver tissue were detected by qRT-PCR and Western blotting. Neutrophil infiltration was observed by Ly6G immunofluorescence staining, and hepatocyte apoptosis was detected by TUNEL staining. Results Compared with R. roxburghii juice, the contents of total acids, total phenols and total flavonoids in R. roxburghii fermentation broth were significantly increased (P < 0.05), while the content of vitamin C was significantly decreased (P < 0.05), and the total amount of major antioxidant active ingredients were increased, and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging rate and Fe³⁺ reducing ability were significantly increased (P < 0.01). Compared with control group, model group showed obvious pathological changes such as hemorrhagic injury in liver, liver index and activities of ALT, AST, levels of inflammatory factors in serum and MDA level in liver tissue were significantly increased (P < 0.001), while GSH level and SOD activity in liver tissue were significantly decreased (P < 0.001). Inflammatory cell infiltration in liver tissue, disruption of hepatic cord structure, and increased necrotic area were observed; The mRNA and protein expression levels of Nrf2, HO-1, SOD-1 and CAT in liver tissue were significantly reduced (P < 0.001); Neutrophil infiltration and hepatocyte apoptosis in liver tissue were significantly increased (P < 0.001). Compared with model group, NAC and R. roxburghii fermentation broth all dose groups could improve the above pathological and biochemical indicators to varying degrees, including reducing visible liver damage and lesions, lowering liver index, down-regulating transaminase activity and inflammatory factor levels in serum, reducing MDA level in liver tissue, increasing GSH level and SOD activity, and up-regulating Nrf2/ARE pathway related genes and protein expressions (P < 0.05, 0.01, 0.001), while significantly inhibiting neutrophil infiltration and liver cell apoptosis (P < 0.05, 0.001). Conclusion R. roxburghii fermentation broth has a potential liver-protective effect against APAP-induced DILI in mice, and its mechanism may be related to the activation of Nrf2/ARE signaling pathway, enhancing antioxidant capacity and inhibiting inflammatory responses.

Rosa roxburghii fermentation broth  /  drug-induced liver injury  /  antioxidant  /  anti-inflammatory  /  Nrf2/ARE signaling pathway  /  vitamin C  /  polyphenols  /  flavonoids