植物活性成分对细胞存活率的影响及其测定方法的比较研究  

Effects of Bioactive Plant Components on Cell Viability and Comparison of Cell Viable Assays

作　　者：孙健[1] 梅晶[1] 赵晓红[1] 刘涛[1] 阮研硕[1] 

机构地区：[1]北京联合大学应用文理学院生物活性物质与功能食品北京市重点实验室,北京100191

出　　处：《食品科学》2012年第19期119-123,共5页Food Science

基　　金：北京市教育委员会科技计划重点项目(KZ201211417041)

摘　　要：选用植物活性物质单体白藜芦醇和绿原酸作用于中国仓鼠卵巢(CHO)细胞,采用MTS法、WST-1法、CCK-8法、XTT法和MTT法测定细胞存活率,并将MTS法、WST-1法、CCK-8法、XTT法这4种方法与传统的MTT法进行比较分析,从中选出最佳的检测方法;通过IC50值比较白藜芦醇和绿原酸毒性大小。结果显示:CCK-8法所得结果稳定,且与MTT法所测得的IC50值最接近,为最佳选择;WST-1法的实验可重复性较差,结果偏小;XTT法是4种方法中最不稳定的,作用时间、检测波长最难确定,且实验重复性差。CCK-8法、MTS法、WST-1法、XTT法和MTT法测定所得白藜芦醇的IC50值分别是116.86、124.68、64.90、162.86、86.30μmol/L;绿原酸的IC50值分别为2808.60、1119.96、1029.72、1325.43、2852.63μmol/L,同种方法下绿原酸的IC50值均高于白藜芦醇的IC50值。结论是测定植物活性成分对细胞存活率影响的方法以CCK-8为最好。白藜芦醇对CHO细胞的毒性比绿原酸大。Resveratrol and chlorogenic acid as phytochemicals were used to treat CHO cells. Cell viability was measured by using MTS, WST-1, CCK-8, XTr and M＇IT assays. Comparison of MTF assay with four others was carried out to screen the optimal cell viability array. Along with this, the cytotoxicity of resveratrol and cholorogenic acid was evaluated by IC50. The CCK-8 assay provided the most similar IC50 to the MTT assay with good stability and consequently was the best choice. In contrast, the WST-1 assay showed worst repeatability and smaller results. The XTT assay was the least stable and less repeatable and the optimal reaction time and detection wavelength was the most difficult to determine. The IC50 values of resveratrol were 116.86, 124.68, 64.90, 162.86 μmol/L and 86.30 μmol/L respectively, and the IC50 values of chlorogenic acid were 2808.60, 1119.96, 1029.72, 1325.43 gmol/L and 2852.63 μmol/L respectively as determined by the CCK-8, MTS, WST-1, XTT and MTr assays. These data indicated that nigher IC50 value could be obtained for chlorogenic acid by each assay compared with resveratrol. In light the above findings, we conclude that the CCK-8 assay provides the best approach to determining the effects of bioactive plant components on cell viability and that resveratrol has nigher cytotoxicity to CHO cells than chlorogenic acid.

关 键 词：MTS WST-1 CCK-8 XTT 细胞存活率 白黎芦醇 绿原酸 

分 类 号：Q946[生物学—植物学] R994.4[医药卫生—毒理学]