诸葛菜种子水提物对硫代乙酰胺诱导小鼠急性肝损伤的保护作用研究  

Protective effect of water extracts of Orychophragmus violaceus seeds on TAA-induced acute liver injury in mice

作　　者：庞敏 单宇 王菲菲 杨淑贤[2] 李立勇[2] 曹丽[2] PANG Min;SHAN Yu;WANG Fei-fei;YANG Shu-xian;LI Li-yong;CAO Li(Pharmaceutical Engineering Technology Research Center,College of Pharmacy,Harbin University of Commerce,Harbin 150076,China;Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100193,China;Guangxi University of Chinese Medicine,Nanning 530001,China)

机构地区：[1]哈尔滨商业大学药学院药物工程技术研究中心,黑龙江哈尔滨150076 [2]中国医学科学院北京协和医学院药用植物研究所,北京100193 [3]广西中医药大学,广西南宁530001

出　　处：《中国中药杂志》2020年第6期1399-1405,共7页China Journal of Chinese Materia Medica

基　　金：北京市自然科学基金青年基金项目(7164281)。

摘　　要：该实验研究了诸葛菜种子水提物对硫代乙酰胺(TAA)诱导小鼠急性肝损伤的保护作用。将雄性ICR小鼠随机分为7组:正常组,模型组,双环醇阳性对照组(200 mg·kg-1)及葵花护肝片阳性对照组(350 mg·kg-1),诸葛菜种子水提物低(125 mg·kg-1)、中(250 mg·kg-1)、高(500 mg·kg-1)剂量组。各组按照0.02 mL·g-1灌胃给药,每日1次,连续给药4 d,第4天给药后1 h,除正常组外均腹腔注射硫代乙酰胺溶液100 mg·kg-1建立急性肝损伤模型,24 h后脱臼处死取血、取材。计算肝脏指数;检测血清中ALT,AST,TBiL活性及肝组织中SOD,GSH-Px的活性,MDA,GSH的含量;HE染色法观察肝组织病理变化;Western blot法检测NF-κB p65,Keap-1,Nrf2,p-p38,p-ERK,p-JNK,Bax,Bcl-2,caspase-3,cleaved caspase-3,caspase-8蛋白的表达。结果表明,与模型组相比,诸葛菜各给药组均能明显改善肝脏病理变化,降低肝损伤小鼠血清中ALT,AST,TBiL活性,其高剂量组肝脏组织匀浆上清中SOD与GSH-Px活性升高,GSH含量升高,MDA含量降低,同时可致肝组织中Bax,Keap-1,p-p38,p-JNK,p-ERK,NF-κB p65,cleaved caspase-3蛋白表达水平减少和Nrf2,Bcl-2,caspase-3,caspase-8蛋白表达水平增加。因此,诸葛菜种子水提物对硫代乙酰胺溶液所致小鼠急性肝损伤有明显的保护作用,其机制可能与下调Keap-1,上调Nrf2蛋白表达减轻氧化应激,抑制p38,ERK的磷酸化水平,进而抑制NF-κB信号通路的激活,抑制JNK蛋白的磷酸化,下调Bax,上调Bcl-2蛋白的表达,抑制肝细胞凋亡有关。This study aimed to investigate the protective effect of water extracts of Orychophragmus violaceus seeds on liver injury induced by thioacetamide(TAA)in mice.ICR male mice were randomly divided into seven groups:normal group,model group,bicyclol positive control group(200 mg·kg-1),Kuihua Hugan Tablets group(350 mg·kg-1),O.violaceus seeds low-dose water extract group(125 mg·kg-1),middle-dose water extract group(250 mg·kg-1),and high-dose water extract group(500 mg·kg-1).Intragastric administration was given in all groups at 0.02 mL·g-1 body weight,1 time a day for continuous 4 days.One h after the administration on the 4 th day,the liver injury model was induced by intraperitoneal injection of TAA(100 mg·kg-1).The mice were put to death 24 hours later.Blood and tissues were taken and organ indexes were calculated.The activities of ALT,AST and TBiL in serum were detected.The content of MDA,GSH and the activity of SOD,GSH-Px in liver homogenate were examined by colorimetry method.HE staining was used to observe the pathological changes of liver tissues in mice.The protein expression levels of NF-κB p65,Keap-1,Nrf2,p-p38,p-JNK,p-ERK,Bax,Bcl-2,caspase-3,cleaved caspase-3 and caspase-8 were detected by Western blot.The results showed that as compared with the model group,various O.violaceus seeds groups could significantly improve the pathological conditions of liver and reduce ALT,AST,TBiL activities in serum of mice with liver injury.In the high-dose group,the activities of SOD,GSH-Px and the content of GSH were significantly increased,while MDA content was sharply declined.Meanwhile,O.violaceus seeds extract down-regulated the expressions of Bax,Keap-1,p-p38,p-JNK,p-ERK,NF-κB p65,cleaved caspase-3 and up-regulated the expressions of Nrf2,Bcl-2,caspase-3 and caspase-8.In conclusion,O.violaceus seeds extract exhibited potent protective effect on liver injury induced by TAA in mice,and its mechanism may be related to down-regulating levels of Keap-1,up-regulating the expressions of Nrf2,inhibiting the express

关 键 词：诸葛菜 硫代乙酰胺 急性肝损伤 保护作用 NF-ΚB 

分 类 号：R285.5[医药卫生—中药学]