论文部分内容阅读
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。
To investigate the protective effect of luteolin (Lut) on L02 hepatocytes injury induced by acetaminophen (APAP). CCK-8 assay was used to detect the effect of Lut on the activity of L02 cells. The concentration and duration of LAP injury induced by APAP were screened by cell morphology, CCK-8 assay and flow cytometry The content of malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) in the cell supernatant were detected by colorimetric assay. The apoptosis-related genes Bax and Bcl-2 , caspase-3 expression. The results showed that Lut at 2.5 ~ 40μmol·L -1 did not affect the activity of L02 cells; 12 mmol·L -1 APAP could be used to establish hepatocytes injury model at 12 hours. Compared with the model group, Lut group significantly improved cell morphology, increased cell body, adherent ability recovered significantly; apoptosis rate decreased significantly; MDA content decreased significantly (P <0.05 or P <0.01), GSH and SOD activity increased significantly (P <0.05 or P <0.01). At the same time, the expression of Bcl-2 and Bax and caspase-3 mRNA were up-regulated (P <0.05 or P <0.01). This experiment demonstrated that Lut has a protective effect on APAP-induced L02 cell injury, and its mechanism may be related to its reduction of oxidative stress and inhibition of apoptosis.