木犀草素对对乙酰氨基酚诱导的L02肝细胞损伤的保护作用  

Protective effects of luteolin against acetaminophen-induced damage in L02 liver cells

作　　者：贺兰芝 孟雅坤 韩延忠 张振芳[2] 尹萍[3,2] 桑秀秀 肖小河[2] 柏兆方[2] 

机构地区：[1]湖南中医药大学药学院,湖南长沙410208 [2]解放军302医院全军中医药研究所,北京100039 [3]江西中医药大学药学院,江西南昌330004

出　　处：《中国中药杂志》2016年第22期4234-4239,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金青年基金项目(81503350);国家“重大新药创制”科技重大专项(2015ZX09501-004-001-008);国家公益性行业科研专项(201507004-04)

摘　　要：探讨木犀草素(luteolin,Lut)对对乙酰氨基酚(acetaminophen,APAP)诱导的L02肝细胞损伤的保护作用。CCK-8法检测Lut对L02细胞活性的影响;筛选APAP诱导L02细胞损伤的浓度及作用时间;细胞形态学、CCK-8实验和流式细胞术检测分析Lut对APAP诱导的L02细胞凋亡的影响;比色法检测细胞上清液中丙二醛(MDA)含量、谷胱甘肽(GSH)及超氧化物歧化酶(SOD)活性;RT-PCR检测凋亡相关基因Bax,Bcl-2,caspase-3的表达。结果显示Lut在2.5~40μmol·L^(-1)不影响L02细胞活性;12 mmol·L-1APAP作用于L02细胞12 h可用于建立肝细胞损伤模型。与模型组相比,Lut组细胞状态明显改善,胞体增大,贴壁能力恢复明显;细胞凋亡率明显下降;MDA含量显著下降(P<0.05或P<0.01),GSH和SOD活性显著提高(P<0.05或P<0.01),同时能够上调Bcl-2及下调Bax,caspase-3 mRNA的表达(P<0.05或P<0.01)。该实验证明了Lut对APAP诱导的L02细胞损伤有保护作用,其机制可能与其减轻氧化应激反应和抑制细胞凋亡有关。This paper was aimed to investigate the protective effects of luteolin （Lut） against aeetaminophen（APAP） -induced damage in L02 liver cells. CCK-8 was used to detect the cell activation of L02 cells treated by different Lut. The concentration and time of APAP induced L02 cell damage was screened. The effect of Lut on APAP induced apoptosis of 1.02 cells was detected by cell morphological observation, CCK-8 assay and flow eytometry. The contents of MDA, GSH and SOD activity in cell supernatant were detected by eolorimetric assay. The expression of apoptosis-related genes Bax, Bcl-2 and caspase-3 was detected by RT-PCR. The results showed that Lut in 2. 5-40 μmol · L^-1 range does not affect the activity of L02 cells; 12 mmol · L^-1 APAP incubated with L02 cell 12 h to establish damage model. Compared with the model group, the cell status of Lut group was significantly improved, the cell body was increased, the adherence ability was recovered, and the apoptosis rate was obviously decreased. MDA content decreased significantly （P 〈 0. 05, P 〈 0.01 ）, GSH and SOD activity significantly increased （ P 〈 0.05, P 〈 0.01 ） , at the same time, it could up-regulate expression of Bcl-2 mRNA and down-regulate the expression of Bax and caspase-3 mRNA. In conclusion, Lut has protective effect on APAP induced L02 cell injury, and its mechanism may be related to the reduction of oxidative stress and inhibition of apoptosis.

关 键 词：木犀草素 对乙酰氨基酚 L02细胞 氧化应激 细胞凋亡 

分 类 号：R285[医药卫生—中药学]