目的 探究刺梨发酵液对对乙酰氨基酚（acetaminophen，APAP）诱导的药物性肝损伤（drug-induced liver injury，DILI）小鼠模型的保护作用，并基于核因子红细胞2相关因子2（nuclear factor erythroid 2-related factor，Nrf2）/抗氧化反应元件（antioxidant response element，ARE）信号通路探讨其潜在机制。方法 以刺梨原汁为对照，分析刺梨发酵液主要活性成分含量及体外抗氧化能力；将60只昆明种小鼠随机分为对照组、模型组、N-乙酰半胱氨酸（N-acetylcysteine，NAC，0.15 g/kg）组及刺梨发酵液低、中、高剂量（5、10、20 mL/kg）组，每组10只。连续给药7 d，末次给药1 h后，除对照组外，其余各组ip APAP（400 mg/kg）以建立急性肝损伤模型。造模12 h后，麻醉并处死小鼠，立即取出小鼠完整肝脏，观察有无肉眼可见损伤及病变；测定小鼠体质量及肝脏质量，计算肝脏指数；采用ELISA检测血清中丙氨酸氨基转移酶（alanine aminotransferase，ALT）、天冬氨酸氨基转移酶（aspartate aminotransferase，AST）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、IL-1β水平及肝组织超氧化物歧化酶（superoxide dismutase，SOD）活性以及谷胱甘肽（glutathione，GSH）、丙二醛（malonaldehyde，MDA）水平；苏木素-伊红（hematoxylin-eosin，HE）染色观察肝组织病理变化；qRT-PCR和Western blotting检测肝组织中Nrf2、血红素氧合酶-1（heme oxygenase-1，HO-1）、SOD-1、过氧化氢酶（catalase，CAT）的mRNA及蛋白表达；Ly6G免疫荧光染色观察中性粒细胞浸润情况；TUNEL染色检测肝细胞凋亡情况。结果 与刺梨原汁相比，刺梨发酵液中总酸、总酚及总黄酮含量显著升高（P＜0.05），维生素C含量显著下降（P＜0.05），主要抗氧化活性成分总量增加，且其1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl，DPPH）自由基清除率与Fe³⁺还原能力均显著增强（P＜0.01）。与对照组比较，模型组小鼠肝脏出现明显的出血性病理损伤，肝脏指数、血清中ALT及AST活性、炎症因子水平和肝组织MDA水平均显著升高（P＜0.001），而肝组织GSH水平和SOD活性显著下降（P＜0.001）；肝组织炎性细胞浸润、肝索结构紊乱及坏死面积增加；肝组织Nrf2、HO-1、SOD-1、CAT的mRNA及蛋白表达水平显著降低（P＜0.001）；肝组织中性粒细胞浸润及肝细胞凋亡显著增加（P＜0.001）。与模型组比较，NAC及刺梨发酵液各剂量组均可不同程度地改善上述病理及生化指标，包括减轻肉眼可见的肝脏损伤及病变，降低肝脏指数，下调血清转氨酶活性及炎症因子水平，降低肝组织MDA水平，提升GSH水平和SOD活性，并上调Nrf2/ARE通路相关基因及蛋白表达（P＜0.05、0.01、0.001），同时显著抑制中性粒细胞浸润与肝细胞凋亡（P＜0.05、0.001）。结论 刺梨发酵液对APAP诱导的DILI小鼠具有潜在的肝保护作用，其机制可能与激活Nrf2/ARE信号通路、增强机体抗氧化能力并抑制炎症反应有关。

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.

R285.5

贵州省中医药管理局中医药、民族医药科学技术课题（QZYY-2025-102）;贵州省科技重大专项项目（黔科合重大专项字[2024]015）