苦丁茶熊果酸对鼻咽癌细胞药物干预作用时间与浓度选择

目的探讨苦丁茶熊果酸对鼻咽癌细胞干预作用的最适宜时间与浓度。方法从苦丁茶老叶中提取熊果酸并纯化与鉴定,用5,10,20,40,80,100μmol-L-1 6种浓度的熊果酸,持续作用于在培养的鼻咽癌细胞24,48,72,96,120 h后,用MTT法检测鼻咽癌细胞被抑制情况,流式细胞术分析其细胞周期和凋亡率;光学显微镜观察细胞形态学变化。结果 40,80μmol-L-1的苦丁茶熊果酸经过24 h的作用,抑制NCE-2细胞分别为（41.21±0.25）%和（94.11±0.37）%,两者均能阻滞鼻咽癌细胞于G0/G1期;其阻滞率分别为（73.41±0.17）%和（75.26±0.18）%,40μmol-L-1苦丁茶熊果酸作用24 h时,鼻咽癌细胞即出现细胞萎缩,细胞黏附能力下降,一些细胞逐渐发生碎裂而死亡,细胞数量明显变少;作用48 h后细胞萎缩,轮廓改变程度更深,呈现部分细胞悬浮,黏附能力丧失,发生碎裂而死亡细胞增多,贴壁存活的细胞数量更少。结论从苦丁茶提取的熊果酸对鼻咽癌细胞有较强的干预作用,药物干预的最适宜作用浓度为40μmol-L-1,药物干预后形态学观察的最适宜时间为24 h。

Exploring the Proper Intervention Time and Concentration of Ursolic Acid of Kuding Tea on Human Nasopharyngeal Carcinoma Cell

OBJECTIVE To explore the proper intervention time and concentration of ursolic acid of Kuding tea on human nasopharyngeal carcinoma cell.METHODS Ursolic acid was extracted from the old leaves of Kuding tea and was then purified and tested.Different concentrations(5,10,20,40,80,100 μmol·L-1) of ursolic acid were used to interfere with human nasopharyngeal carcinoma cell for different lengths of time(24,48,72,96,120 h).MTT method was used in determining the effect of ursolic acid on cell proliferation inhibition.Microscopy method was used to observe cytotoxicity of ursolic acid.Flow cytometer method was used to determine apoptosis induction and cell cycle blocking effect of ursolic acid.RESULTS With 24 h of intervention time,the inhibitory rates of 40 and 80 μmol-L-1 of ursolic acid of Kuding tea on NEC-2 cells were(41.21±0.25)% and(94.11±0.37)%,respectively;and both applications could block cells at G0/G1 phases with blocking rate of(73.41±0.17)% and(75.26±0.18)%,respectively.After 24 h intervention of 40 μmol·L-1 of ursolic acid of Kuding tea,cell atrophy appeared,cell adhesion decreased,some cells died from fragmentations and cell’s number was obviously reduced.After 48 h intervention of 40 μmol·L-1 of ursolic acid of Kuding tea,these effects were further strengthened and became more obvious.CONCLUSION The ursolic acid of Kuding tea has strong intervention on human nasopharyngeal carcinoma cell.The concentration of 40 μmol·L-1 and the observation time of 24 h are recommended in application.