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 control group and LHRD treatment groups(P<0.01).Morphological changes of liver tissues were also alleviated.Moreover,liver GSH content(P<0.01,P<0.05)and GCLC protein expression(P<0.01)were significantly up-regulated.Conclusion LHRD had protective effect on ALI induced by excessive acetaminophen,and the mechanism was possibly related to enhancing capacities of antioxidant stress to protect liver cells.
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)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
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)。
