该实验以小鼠正常肝细胞AML12为模型，为了研究番茄红素对棒曲霉素诱导的细胞凋亡的抑制作用。不同方式处理细胞后，采用CCK-8检测细胞存活率，采用乳酸脱氢酶释放检测番茄红素对细胞膜的保护作用，检测ROS水平及SOD水平，通过荧光显微镜观察细胞形态学的变化，Annexin V-EGFP/PI标记检测细胞凋亡率，Western Blot测定凋亡相关蛋白Chop、Grp78、Caspase-12、Caspase-9、Caspase-3、Bcl-2和Bax的表达水平。CCK-8结果显示，番茄红素可显著抑制棒曲霉素诱导的AML12细胞凋亡。6 μmol?L-1 PAT处理后的细胞存活率为57.09%。先用LYC孵育24 h，再用6 μmol?L-1 PAT处理细胞存活率为73.84%。番茄红素预处理细胞LDH释放量较棒曲霉素单独孵育降低29.25%，番茄红素能有效保护细胞膜完整性。AV/PI结果显示，棒曲霉素诱导细胞凋亡和坏死率为38.24%，加入番茄红素诱导细胞凋亡和坏死率为25.57%，晚期凋亡细胞降低9.37%。细胞氧化还原系统指标的检测显示，番茄红素能抑制棒曲霉素造成的SOD活力下降，进而减轻ROS过量。结果表明，番茄红素通过调节内质网应激途径显著抑制棒曲霉素诱导的小鼠正常肝细胞AML12凋亡。该研究为番茄红素的肝保护机制提供了理论基础。

Using mouse normal liver cells AML12 as a model, the inhibitory effect of lycopene on patulin-induced apoptosis was studied in this experiment. After the cells were treated in different ways, the cell viability was detected by CCK-8 assay, the protective effect of lycopene on cell membrane was detected by lactate dehydrogenase (LDH) release assay, the levels of reactive oxygen species (ROS) and superoxide dismutase (SOD) were measured, the morphological changes of cells were observed by fluorescence microscopy, the apoptosis rate was detected by Annexin V-EGFP/PI staining, and the expression levels of apoptosis-related proteins such as Chop, Grp78, Caspase-12, Caspase-9, Caspase-3, Bcl-2 and Bax were determined by Western Blot. It was shown by the results of CCK-8 assay that patulin-induced apoptosis of AML12 cells could be significantly inhibited by lycopene. The cell viability was 57.09% after being treated with 6 μmol?L-1 patulin. When the cells were incubated with lycopene for 24 hours first and then treated with 6 μmol?L-1 patulin, the cell viability was 73.84. The release amount of LDH in cells pretreated with lycopene was 29.25% lower than that of cells incubated with patulin alone, which indicated that the integrity of cell membrane could be effectively protected by lycopene. It was shown by the results of Annexin V-EGFP/PI staining that the apoptosis and necrosis rates of cells induced by patulin were 38.24%, while those were 25.57% after adding lycopene, and the rate of late apoptotic cells decreased by 9.37%. It was shown by the detection of cell redox system indexes that the decline of SOD activity caused by patulin could be inhibited by lycopene, thereby reducing the excessive production of ROS. It was indicated by the results that patulin-induced apoptosis of mouse normal liver cells AML12 could be significantly inhibited by lycopene by regulating the endoplasmic reticulum stress pathway. This research furnishes a theoretical foundation for the hepatoprotective mechanism of lycopene.

国家自然科学基金项目（面上项目，重点项目，重大项目）