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目的研究艾塞那肽(EXE)是否通过抑制内质网应激来减少棕榈酸诱导的肝细胞凋亡。方法将人肝细胞株L-02分为对照组、模型组(棕榈酸500μmol·L~(-1)处理24 h)、3个剂量实验组(以棕榈酸500μmol·L~(-1)和EXE 25,50,100 nmol·L~(-1)分别处理24 h)和联合组(以棕榈酸500μmol·L~(-1)、EXE 100nmol·L~(-1)和衣霉素2μg·L~(-1)处理24 h)。用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞活力;以脱氧核苷酸末端转移酶介导的dUTP缺口末端标记(TUNEL)染色及半胱天冬酶(Caspase)-3活性检测观察细胞凋亡;以实时荧光定量聚合酶链式反应(RT-PCR)法检测内质网应激标志物C/EBP同源蛋白(CHOP)表达及Caspase-12活性以观察细胞内质网应激水平。结果与模型组比较,EXE能剂量依赖性增加棕榈酸处理后L-02细胞的活力,减少凋亡细胞数,降低Caspase-3和Caspase-12活性,并下调CHOP表达(均P<0.05)。与高剂量实验组比较,联合组的CHOP表达、细胞凋亡数及Caspase-12活性均显著升高,细胞活力显著下降[(78.58±1.78)%vs(87.78±3.66)%](均P<0.05)。结论 EXE可能通过抑制内质网应激减少脂肪酸诱导肝细胞凋亡。
Aim To investigate if exenatide (EXE) reduces palmitate-induced hepatocyte apoptosis by inhibiting endoplasmic reticulum stress. Methods Human hepatocellular carcinoma cell line L-02 was divided into control group, model group (treated with 500μmol·L -1 palmitate for 24 hours), three dose groups (500μmol·L -1 palmitate and EXE 25, 50 and 100 nmol·L -1 for 24 h) and the combination group (500 μmol·L -1 palmitate, 100 nmol·L -1 EXT and 2 μg · L -1) (-1) for 24 h). Cell viability was measured by 3- (4,5-dimethylthiazol-2) -2,5-diphenyltetrazolium bromide (MTT) assay. Deoxyribonucleotide transferase mediated dUTP nick end (TUNEL) staining and Caspase-3 activity assay were used to observe the cell apoptosis. The endoplasmic reticulum stress index C / EBP was detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) Source protein (CHOP) expression and Caspase-12 activity in order to observe the cellular endoplasmic reticulum stress. Results Compared with the model group, EXE could increase the activity of L-02 cells in a dose-dependent manner, decrease the number of apoptotic cells, decrease the activity of Caspase-3 and Caspase-12 and down-regulate the expression of CHOP (all P <0.05). Compared with the high dose group, the CHOP expression, the number of apoptotic cells and the activity of Caspase-12 in the combined group were significantly increased (P <0.05) 0.05). Conclusion EXE may reduce fatty acid-induced hepatocyte apoptosis by inhibiting endoplasmic reticulum stress.