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目的了解顺铂压力下人肝母细胞瘤株(HepG2)细胞内活性氧(ROS)的水平及线粒体膜电位的变化及意义。方法低剂量顺铂与HepG2细胞共孵。活性氧探针(2,7-二氯荧光素)探测顺铂压力下HepG2细胞内活性氧水平的变化;用反应膜电位荧光探针罗丹明-123,在激光扫描共聚焦显微镜下, 通过荧光强度反映线粒体膜电位的变化。于0、24、48、72、120、168 h动态观察HepG2细胞内活性氧和线粒体膜电位的变化。结果顺铂持续作用24 h和48 h时,活性氧的产生较正常明显增多,尤以24 h时最明显,72 h后开始下降,120 h后基本接近正常;顺铂选择性压力下,24 h和48 h HepG2细胞内罗丹明123的荧光强度明显减弱,以24 h时最明显,72 h以后逐渐升高,120 h以后与未经处理的HepG2细胞无明显差异。结论HepG2细胞在低剂量顺铂的压力下,细胞内活性氧水平和线粒体膜电位的变化,提示HepG2 细胞可对低剂量顺铂的持续压力发生适应性变化。
Objective To investigate the level of reactive oxygen species (ROS) and the mitochondrial membrane potential and its significance in human hepatoblastoma cell line HepG2 under cisplatin pressure. Methods Low-dose cisplatin was incubated with HepG2 cells. Reactive oxygen species probe (2,7-dichlorofluorescein) to detect changes in the level of reactive oxygen species in HepG2 cells under cisplatin pressure; using a reactive membrane potential fluorescent probe Rhodamine-123 under a laser scanning confocal microscope, Intensity reflects changes in mitochondrial membrane potential. The changes of reactive oxygen species and mitochondrial membrane potential in HepG2 cells were observed at 0, 24, 48, 72, 120 and 168 h. RESULTS: After 24 and 48 hours of cisplatin treatment, the production of reactive oxygen species (ROS) was significantly higher than that of normal group, especially at 24 hours and then decreased at 72 hours, and nearly normal after 120 hours. Under the selective pressure of cisplatin 24 Fluorescence intensity of rhodamine 123 in HepG2 cells was significantly decreased at 48 h and 48 h, which was the most obvious at 24 h and gradually increased after 72 h. There was no significant difference between HepG2 cells and untreated HepG2 cells after 120 h. Conclusion The changes of intracellular reactive oxygen species (ROS) and mitochondrial membrane potential in HepG2 cells under the low dose of cisplatin suggest that HepG2 cells can adaptively change the sustained pressure of low-dose cisplatin.