独活醇提物对四氯化碳诱导小鼠急性肝损伤的保护作用及机制  

Protective effect and mechanism of ethanol extract of Angelicae Pubescentis Radix on carbon tetrachloride-induced acute liver injury in mice

作　　者：吴诗慧 龙潇云 赵邯涛 袁吕江[1,2] 陈前锋[1,2] WU Shihui;LONG Xiaoyun;ZHAO Hantao;YUAN Lyujiang;CHEN Qianfeng(School of Pharmaceutical Sciences,Southwest University,Chongqing 400715,China;Engineering Research Center of the Ministry of Education for the Development and Utilization of Coptis chinensis,Southwest University,Chongqing 400715,China)

机构地区：[1]西南大学药学院,重庆400715 [2]西南大学黄连开发与利用教育部工程研究中心,重庆400715

出　　处：《中草药》2024年第17期5867-5874,共8页Chinese Traditional and Herbal Drugs

基　　金：重庆市中医药重点建设学科(2021-4322190044)。

摘　　要：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)�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-1βin 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-1βin 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 sign

关 键 词：独活 急性肝损伤 氧化应激 炎症 蛇床子素 二氢欧山芹醇当归酸酯 Nrf2/HO-1信号通路 TLR4/MyD88/NF-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]