目的 研究独活醇提物对四氯化碳（carbon tetrachloride，CCl₄）致小鼠急性肝损伤的保护作用及作用机制。方法 60只雄性昆明小鼠随机分为对照组、模型组、水飞蓟宾（63 mg/kg）组和独活醇提物低、中、高剂量（50、150、300 mg/kg）组，每组10只。连续给药1周，于末次给药2 h后ip 0.2% CCl₄溶液致小鼠急性肝损伤，24 h后收集血清和肝脏组织。采用试剂盒检测小鼠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）、过氧化物歧化酶（superoxide dismutase，SOD）活性和谷胱甘肽（glutathione，GSH）含量；采用试剂盒检测肝组织中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）和IL-1β水平；采用苏木素-伊红（hematoxylin-eosin，HE）染色法观察肝组织病理变化；采用Western blotting检测肝组织Toll样受体4（Toll-like receptor 4，TLR4）、髓分化因子88（myeloid differentiation factor 88，MyD88）、磷酸化κB抑制蛋白激酶（phosphorylated kappa B inhibitor protein kinase，p-IKK）、磷酸化IκB-α抑制蛋白（phosphorylated kappa B alpha kinase，p-IκBα）、磷酸化核因子-κB p65（phosphorylated nuclear factor-κB p65，p-p65）、TNF-α、核因子E2相关因子（nuclear factor erythroid-derived factor 2-related factor，Nrf2）和血红素氧合酶-1（heme oxygenase-1，HO-1）蛋白表达。结果 与对照组比较，模型组小鼠血清中AST、ALT活性显著升高（P＜0.001），GSH水平和SOD活性显著下降（P＜0.01），肝组织TNF-α、IL-6、IL-1β水平显著升高（P＜0.001），肝组织中Nrf2/HO-1和TLR4/MyD88/NF-κB信号通路相关蛋白表达显著上调（P＜0.05、0.01）。与模型组比较，独活醇提物组小鼠肝组织病理损伤明显改善，血清中AST、ALT活性显著降低（P＜0.01），GSH水平和SOD活性均显著升高（P＜0.01、0.001），肝组织TNF-α、IL-6、IL-1β水平显著降低（P＜0.01、0.001），肝组织中Nrf2/HO-1和TLR4/MyD88/NF-κB信号通路相关蛋白表达显著下调（P＜0.05、0.01）。结论 独活醇提物对CCl₄所致小鼠急性肝损伤具有明显的保护作用，其作用机制可能为通过激活Nrf2/HO-1信号通路抗氧化损伤，同时调控TLR4/MyD88/NF-κB信号通路减轻炎症反应，从而发挥其保肝作用。

Objective To study the protective effect and mechanism of ethanol extract of Duhuo (Angelicae Pubescentis Radix) against acute liver injury in mice induced by carbon tetrachloride (CCl₄). Methods A total of 60 male Kunming mice were randomly divided into control group, model group, silybin (63 mg/kg), ethanol extract of Angelicae Pubescentis Radix low-, medium- and high-dose (50, 150, 300 mg/kg) groups, with 10 mice in each group. Drugs were continuously administered for one week, acute liver injury was induced in mice by intraperitoneally injection of 0.2% CCl₄ solution 2 h after the final administration. After intraperitoneally injection for 24 h, serum and liver tissue were collected. The activities of alanine aminotransferase (ALT), aspartate transaminase (AST), superoxide dismutase (SOD) and conten of glutathione (GSH) in serum were detected by kits; The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in liver tissue were detected by kits; Hematoxylin-eosin (HE) staining was used to observe the pathological changes in liver tissue; Western blotting was used to detect the protein expressions of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), phosphorylated kappa B inhibitor protein kinase (p-IKK), phosphorylated kappa B alpha kinase (p-IκBα), phosphorylated nuclear factor-κB p65 (p-p65), TNF-α, nuclear factor erythroid-derived factor 2-related factor (Nrf2) and heme oxygenase-1 (HO-1) in liver tissue. Results Compared with control group, AST and ALT activities in serum of mice in model group were significantly increased (P < 0.001), GSH level and SOD activity were significantly decreased (P < 0.01), and the levels of TNF-α, IL-6 and IL-1β in liver tissue were significantly increased (P < 0.001), Nrf2/HO-1 and TLR4/MyD88/NF-κB signaling pathway related protein expressions in liver tissue were significantly up-regulated (P < 0.05, 0.01). Compared with model group, liver tissue pathological damage of mice in ethanol extract of Angelicae Pubescentis Radix group was significantly improved, AST and ALT activities in serum were significantly decreased (P < 0.01), GSH level and SOD activity were significantly increased (P < 0.01, 0.001), and the levels of TNF-α, IL-6 and IL-1β in liver tissue were significantly decreased (P < 0.01, 0.001), Nrf2/HO-1 and TLR4/MyD88/NF-κB signaling pathway related protein expressions in liver tissue were significantly down-regulated (P < 0.05, 0.01). Conclusion Ethanol extract of Angelicae Pubescentis Radix has a significant protective effect on CCl₄-induced acute liver injury in mice, and its mechanism may be activating the Nrf2/HO-1 signaling pathway to resist oxidative damage, while regulating the TLR4/MyD88/NF-κB signaling pathway to alleviate inflammatory response, thereby exerting its hepatoprotective effect.

R285.5

重庆市中医药重点建设学科（2021-4322190044）