丹酚酸B对乙醇诱导的HepG2细胞损伤的保护作用  

Protective effects of Salvianolic acid B on HepG2 cells with alcoholic injury

作　　者：邱璐璐[1] 初君怡 张宁[1] QIU Lu-lu;CHU Jun-yi;ZHANG Ning(Department of Pharmacy,the Second Hospital of Dalian Medical University,Dalian 116023,Liaoning Province,China;Department of Pharmacology,Dalian Medical University,Dalian 116044,Liaoning Province,China)

机构地区：[1]大连医科大学附属第二医院药学部,辽宁大连116023 [2]大连医科大学药学院,辽宁大连116044

出　　处：《中国临床药理学杂志》2020年第22期3643-3646,共4页The Chinese Journal of Clinical Pharmacology

基　　金：辽宁省自然科学基金资助项目(20170540257)。

摘　　要：目的通过HepG2细胞体外实验探讨丹酚酸B通过调控AMPKα1/SIRT1通路减轻乙醇损伤的分子机制。方法实验1:HepG2细胞分为空白组、模型组、模型给药组、模型转染组、模型转染给药组。模型转染组和模型转染给药组转染AMPKα1 siRNA,模型给药组和转染后的模型转染给药组加入8μmol·L^-1丹酚酸B,其他组加入等体积0.9%NaCl,3 h后,除空白组,其余4组加入100 mmol·L^-1乙醇诱导48 h造模。实验2:HepG2细胞分为空白组、空白给药组、模型组、模型给药组。给药组加入8μmol·L^-1丹酚酸B,空白组和模型组加入等体积0.9%NaCl,3 h后模型组和模型给药组加入100 mmol·L^-1乙醇后孵育48h造模。实验3:HepG2细胞分为空白组、空白给药组、转染组和转染给药组。转染组和转染给药组转染AMPKα1 siRNA,空白给药组和转染给药组加入8μmol·L^-1丹酚酸B,空白组和转染组加等体积0.9%NaCl。以MTT实验测定细胞存活率,以蛋白质印迹法检测AMPKα1、p-AMPKα1、SIRT1蛋白表达水平。结果实验1:空白组、模型组、模型给药组、模型转染组、模型转染给药组的细胞存活率分别为(100.00±1.58)%,(62.21±2.25)%,(81.65±4.16)%,(58.34±3.11)%,(61.64±3.24)%。模型组和空白组相比,模型给药组和模型组相比,模型转染组和模型组相比,差异均有统计学意义(均P<0.01)。实验2:空白组、空白给药组、模型组、模型给药组细胞中p-AMPKα1/AMPKα1的蛋白表达相对量分别为1.00±0.02,1.35±0.13,0.26±0.08,0.71±0.01,空白给药组与空白组相比,模型组与空白组相比,模型给药组与模型组相比,差异均有统计学意义(P<0.01)。实验3:空白组、空白给药组、转染组、转染给药组细胞中p-AMPKα1/AMPKα1的蛋白表达相对量分别为1.00±0.01,2.31±0.15,0.56±0.05,0.60±0.04,SIRT1的蛋白表达相对量分别为1.00±0.02,2.14±0.20,0.43±0.06,0.46±0.04。空白给药组和空白组相比,转染组和空白组相比,差异均有统计�Objective To investigate the involvement of AMPKα1/SIRT1 pathway in ethanol-induced HepG2 cells injury and to explore the protective mechanisms of Salvianolic acid B(SalB). Methods Experiment 1: The HepG2 cells were divided into five groups: blank group, model group, model administration group, model transfected group and model transfected administration group. After AMPKα1 siRNA was transfected to model transfected group and model transfected administration group,model administration group and model transfected administration group were added 8 μmol·L^-1 SalB,and other groups were added the same amount of normal saline. Three hours later,the HepG2 cells were exposed to ethanol(100 mmol· L^-1) for 48 h except blank group. Experiment 2: The HepG2 cells were divided into four groups: blank group,blank administration group,model group and model administration group. Administration groups were given 8 μmol·L^-1 SalB;blank group and model group were given the same amount of normal saline. Three hours later,model group and model administration group were exposed to ethanol(100 mmol·L^-1) for another 48 h.Experiment 3: The HepG2 cells were divided into four groups: blank group,blank administration group,transfected group and transfected administration group. After transfected group and transfected administration group were transfected with AMPKα1 siRNA,administration groups were given SalB of 8 μmol·L^-1,and blank group and transfected group were given the same amount of normal saline. Cell survival rate were detected. The expressions of AMPKα1,p-AMPKα1 and SIRT1 were detected by Western blotting. Results Experiment 1: The cell survival rate in blank group, model group, model administration group, model transfected group and model transfected administration group were(100. 00 ± 1. 58) %,(62. 21 ± 2. 25) %,(81. 65 ± 4. 16) %,(58. 34 ± 3. 11) %,(61. 64 ± 3. 24) %,respectively. There were statistically significant differences between model group and blank group,model administration group and model group,m

关 键 词：HEPG2细胞 丹酚酸B 沉默信息调节因子2相关酶Ⅰ 丝氨酸/苏氨酸蛋白激酶组成的异源三聚体依赖的蛋白激酶α1 

分 类 号：R28[医药卫生—中药学]