麦角甾苷对急性肝衰竭小鼠肝功能和肝细胞自噬、凋亡的影响  

Effects of acteoside on liver function and autophagy and apoptosis of hepatocytes in mice with acute liver failure

作　　者：魏思灿 陈伟文[1] 黄鸣恋 蔡荣达 陈亚峥 林天来 WEI Si-can;CHEN Wei-wen;HUANG Ming-lian;CAI Rong-da;CHEN Ya-zheng;LIN Tian-lai(Department of Critical Care Medicine,Quanzhou First Hospital,Quanzhou 362000,Fujian Province,China)

机构地区：[1]泉州市第一医院重症医学科,福建泉州362000

出　　处：《中国临床药理学杂志》2022年第14期1668-1672,1677,共6页The Chinese Journal of Clinical Pharmacology

摘　　要：目的研究麦角甾苷对急性肝衰竭小鼠肝功能和肝细胞自噬及凋亡的影响。方法将32只小鼠随机分为对照组、模型组、麦角甾苷低剂量组(20 mg·kg^(-1))和麦角甾苷高剂量组(40 mg·kg^(-1)),每组8只小鼠。对照组小鼠腹腔注射生理盐水200μL;模型组小鼠腹腔注射D-氨基半乳糖盐酸盐(D-GalN)700 mg·kg^(-1)和脂多糖(LPS)10 mg·kg^(-1)诱导急性肝功能衰竭(ALF)小鼠模型;麦角甾苷低剂量组和麦角甾苷高剂量组小鼠在D-GalN/LPS给药前2 h,分别通过腹腔注射20和40 mg·kg^(-1)麦角甾苷。培养NCTC1469细胞,随机分为对照组、模型组、麦角甾苷低剂量实验组和麦角甾苷高剂量实验组。对照组细胞使用正常培养基培养;模型组细胞使用正常培养基培养2 h后加入40 mmol·L^(-1)的D-GalN处理12 h;麦角甾苷低浓度实验组和麦角甾苷高浓度实验组分别以50,100 mol·L^(-1)麦角甾苷干预培养2 h,再加入40 mmol·L^(-1)的D-GalN处理12 h。用全自动生化分析仪检测血清谷丙转氨酶(GPT)和谷草转氨酶(GOT)水平;用酶联免疫吸附(ELISA)试剂盒检测各组小鼠血清中白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平;以苏木精-伊红(HE)染色法观察各组小鼠肝组织病理学情况;以流式细胞仪检测NCTC1469细胞凋亡情况;以蛋白质印迹法检测NCTC1469细胞凋亡和自噬相关蛋白的表达情况。结果对照组、模型组和麦角甾苷低、高剂量组小鼠血清GPT含量分别为(20.42±5.90),(1496.98±108.12),(1048.99±103.26),(304.87±24.79)U·L^(-1),TNF-α含量分别为(24.12±2.72),(196.69±16.66),(88.96±8.76),(45.64±5.76)ng·mL^(-1),TUNEL标记的阳性细胞率分别为(2.02±0.02)%,(46.29±6.87)%,(25.06±3.01)%和(19.03±1.07)%。与模型组相比,不同剂量麦角甾作用后肝组织病理损伤程度减轻,Bcl-2蛋白和p62蛋白水平以明显升高(均P<0.05),而Bax和Cl-caspase-3蛋白和Beclin^(-1)蛋白的表达水平明显降低(均P<0.05)。结论麦角甾苷能够�Objective To investigate the effect of acteoside on liver function,autophagy and apoptosis of hepatocytes in mice with acute liver failure(ALF).Methods The 32 mice were randomly divided into control group,model group,acteoside low-dose group and acteoside high-dose group,with eight mice in each group.The mice in control group were intraperitoneally injected with normal saline 200μL.The mice in model group were intraperitoneally injected with the same amount of 700 mg·kg^(-1) D-galactosamine hydrochloride(D-GalN)and 10 mg·kg^(-1) lipopolysaccharide(LPS)to induce acute liver failure mouse model.Mice in acteoside low-dose group and acteoside high-dose group were intervened by intraperitoneal injection of 20 and 40 mg·kg^(-1) acteoside,respectively,at 2 h before D-GalN/LPS administration.The NCTC1469 cells were cultured and randomly divided into control group,model group,acteoside low-dose group and acteoside high-dose group.The cells in control group were cultured in normal medium.The cells in model group were cultured in normal medium for 2 h and then treated with 40 mmol·L^(-1) D-GalN for 12 h.The rats in acteoside low-dose group and acteoside high-dose group were incubated with 50 and 100 mol·L^(-1) acteoside for 2 h,respectively,and then treated with 40 mmol·L^(-1) D-GalN for 12 h.Glutamic-pyruvic transaminase(GPT)and glutamic-oxaloacetic transaminase(GOT)levels were measured by automated biochemical analyzer.The levels of serum interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).Hematoxylin and eosin(HE)staining was used to observe the liver histopathology of mice in each group.D-GalN-induced apoptosis of NCTC1469 cells was detected by flow cytometry.The D-GalN-induced apoptosis in NCTC1469 cells and the expression of autophagy-related proteins were detected by Western blot.Results The mouse serum GPT contents in control group,model group,acteoside low-dose group and acteoside high-dose group were(20.42±5.90),(1496.98±108.12),(1048.99±103.26

关 键 词：麦角甾苷 急性肝衰竭 自噬 凋亡 

分 类 号：R28[医药卫生—中药学]