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本研究以19种可食用硫醚类香料为原料,在体外考察其对人白血病细胞HL-60的细胞毒性作用,比较分析其构效关系,并初步探究其肿瘤细胞毒性机理。用19种硫醚类香料处理HL-60细胞,通过DNA片段化来考察其对HL-60细胞的诱导凋亡作用,并对其进行筛选。结果显示,二糠基二硫醚(DFDS)、双-(2-甲基-3-呋喃基)二硫醚(BMFDS)及甲基2-甲基-3-呋喃基二硫醚(MMFDS)具有很强的细胞毒性。利用CCK-8试剂盒检测,添加10μM上述三种硫醚类香料使HL-60细胞的生存率分别下降41.77±1.18%、53.49±2.88%和23.63±0.90%。通过分析19种硫醚类香料的构效关系,明确硫原子数和呋喃基对硫醚类香料的HL-60细胞毒性起重要作用。为了探讨三种硫醚类香料的细胞毒性机理,采用抗氧化剂谷胱甘肽、N-乙酰半胱氨酸预处理细胞,进一步检测硫醚类香料对细胞生存率和DNA片段化的影响。结果表明,抗氧化剂对BMFDS及DFDS诱导的HL-60细胞毒性具有显著的保护作用,说明这两种硫醚类香料可能通过诱导活性氧产生而发挥细胞毒性作用。
In this study, 19 kinds of edible thioether spices as raw materials, in vitro study of its cytotoxicity on human leukemia cells HL-60, comparative analysis of its structure-activity relationship, and to explore the mechanism of tumor cytotoxicity. HL-60 cells were treated with 19 kinds of thioether-like spices, and their apoptosis induced by HL-60 cells was investigated by DNA fragmentation and screened. The results showed that the activities of DFDS, BMFDS and MMFDS were higher than those of the control. Has a strong cytotoxicity. The survival rate of HL-60 cells decreased by 41.77 ± 1.18%, 53.49 ± 2.88% and 23.63 ± 0.90% respectively by using CCK-8 kit and adding 10μM thioether flavors. Through the analysis of the structure-activity relationship of 19 kinds of thioether flavors, it is clear that the number of sulfur atoms and furyl play an important role in the cytotoxicity of thioether-based fragrance HL-60. In order to investigate the cytotoxicity mechanism of three kinds of thioether flavors, antioxidants such as glutathione and N-acetylcysteine were used to pretreat cells to further investigate the effects of thioether flavors on cell viability and DNA fragmentation. The results showed that antioxidants have a significant protective effect on HL-60 cytotoxicity induced by BMFDS and DFDS, indicating that these two thioether flavors may play a cytotoxic role by inducing reactive oxygen species production.