龙虎人丹防治过量对乙酰氨基酚诱导的小鼠急性肝损伤的作用研究  

Longhu Rendan Alleviated Overdose Acetaminophen-induced Acute Liver Injury in Mice

作　　者：汪午屏 董嘉乐 丁礼琴 金家骅 兰天 WANG Wu-ping;DONG Jia-le;DING Li-qin;JIN Jia-hua;LAN Tian(School of Pharmacy,Guangdong Pharmaceutical University,Guangzhou,510006;Research and Development Department,Shanghai Zhonghua Pharmaceutical Co.,Ltd.,Shanghai,201707;Chinese Medicine Research Institute,Guangdong Pharmaceutical University,Guangzhou,510006)

机构地区：[1]广东药科大学药学院药理系,广州510006 [2]上海中华药业有限公司研发部,上海201707 [3]广东药科大学中医药研究院,广州510006

出　　处：《中国中西医结合杂志》2020年第10期1214-1219,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金面上项目(No.81870420);广东药科大学-上海中华药业有限公司校企合作项目(No.HTDJ2017124)。

摘　　要：目的观察龙虎人丹(LHRD)对过量对乙酰氨基酚(APAP)所致急性肝损伤的防治作用,探讨其防治药物性肝损伤的潜在分子机制。方法取30只C57BL/6J小鼠随机分成正常对照组、APAP模型组、N-乙酰-L-半胱氨酸(NAC)阳性对照组(50 mg/kg)、LHRD低剂量(80 mg/kg)组和LHRD高剂量(160 mg/kg)组,固定时间灌胃给予相应药物6天,其中正常对照组和APAP模型组灌胃给予羧甲基纤维素钠(CMC-Na)。最后一次给药后所有小鼠禁食22 h,腹腔注射250 mg/kg APAP,其中正常对照组腹腔注射PBS,6 h后处死小鼠并取其血清和肝脏。采用试剂盒检测各组小鼠血清谷丙转氨酶(ALT)活性及肝脏中谷胱甘肽(GSH)含量;HE染色观察肝脏组织形态学变化;荧光探针DCFH-DA法检测组织细胞内活性氧(ROS)表达;Western Blot检测肝脏组织谷氨酸-半胱氨酸连接酶催化亚基(GCLC)和核酸内切酶G(EndoG)表达。结果与正常对照组比较,APAP模型组血清ALT活性显著升高(P<0.01),细胞损伤显著加重,EndoG蛋白表达和ROS含量显著增加(P<0.01),肝脏GSH含量及GCLC蛋白表达均下调(P<0.05);与APAP模型组比较,NAC阳性对照组及LHRD给药组血清ALT活性下降(P<0.05),ROS含量及EndoG蛋白表达下调(P<0.01,P<0.05),肝组织形态均有改善,且肝脏GSH含量及GCLC蛋白表达量均显著上调(P<0.01,P<0.05)。结论 LHRD可改善过量对乙酰氨基酚诱导的急性肝损伤,其机制可能与增强肝脏抗氧化应激能力,保护肝细胞有关。Objective To observe the prevention and treatment of Longhu Rendan(LHRD)on acute liver injury(ALI)induced by acetaminophen(APAP)overdose,and to study potential molecular mechanisms for its prevention and treatment.Methods Thirty C57 BL/6 J mice were randomly divided into normal control group,APAP model group,N-acetyl-cysteine(NAC)positive control group(50 mg/kg),LHRD-low dose group(80 mg/kg),and LHRD-high dose group(160 mg/kg),6 in each group.Corresponding drugs were administered to mice by gastrogavage at a fixed time for 6 successive days.Sodium carboxymethyl cellulose(CMC-Na)was administered to mice by gastrogavage to mice in the normal control group and the APAP model group.After the last administration,all mice were fasted for 22 h and APAP 250 mg/kg was intraperitoneally administered to them.PBS was intraperitoneally injected to mice in the normal control group.Mice were harvested after 6 h,serum and livers were collected for biochemical,histological,and immunological analyses.Serum alanine aminotransferase(ALT)activity and liver glutathione(GSH)level were detected according to the manufacturer’s instructions.Hematoxylin-eosin(HE)staining was performed to observe histomorphological changes of liver tissues.DCFH-DA,a fluorescence sensor for reactive oxygen species(ROS),was used to measure the ROS level in liver tissue.In addition,the expression of glutamate-cysteine ligase catalytic subunit(GCLC)and endonuclease G(EndoG)in liver tissues were detected by Western Blot.Results Compared with the normal control group,serum ALT activity in the APAP model group significantly increased(P<0.01).HE staining showed hepatocyte damage was significantly aggravated.Meanwhile,ROS content and EndoG protein expression(P<0.01)significantly increased.In addition,liver GSH level and GCLC protein expression were down-regulated(P<0.05).Compared with the model group,serum ALT activity was down-regulated(P<0.01,P<0.05),as well as the ROS level and EndoG protein expression were down-regulated to different degrees in NAC positive co

关 键 词：龙虎人丹 对乙酰氨基酚 急性肝损伤 氧化应激 

分 类 号：R285.5[医药卫生—中药学]