佛手挥发油对B16黑色素瘤细胞体外增殖的抑制作用  

Inhibitory Effect of Fingered Citron Essential Oil on Proliferation of B16 Melanoma Cells in Vitro

作　　者：吕学维[1] 邵邻相[1] 张均平[1] 麻艳芳[1] 邓刚[1] 徐玲玲[1] 李美[1] 

机构地区：[1]浙江师范大学化学与生命科学学院,金华321004

出　　处：《中国粮油学报》2011年第8期50-54,共5页Journal of the Chinese Cereals and Oils Association

摘　　要：利用水蒸气蒸馏法提取佛手挥发油,MTT法测定细胞活力;倒置显微镜,Hoechst 33342/PI荧光染色及扫描电镜观察细胞凋亡与坏死的形态变化;彗星电泳法检测细胞DNA损伤程度。结果表明:佛手挥发油能够显著抑制B16黑色素瘤细胞增殖(P<0.05),呈剂量依赖效应;佛手挥发油处理B16黑色素瘤细胞6 h后,倒置显微镜下观察到低、中剂量组(62.5、125、250μg/mL)细胞体积缩小,形态变得不规则,细胞碎片增多,部分细胞成团生长,而高剂量组(500μg/mL)细胞膜崩解,胞浆外溢,细胞碎片增多;荧光显微镜下可见低、中剂量组细胞核固缩,核染色质凝集,边缘化等典型细胞凋亡特征,而高剂量组细胞核为PI染成红色,核染色质分布均匀,呈细胞坏死特征;扫描电镜观察到试验组细胞形态不规则,细胞表面不平整,微绒毛减少等细胞形态变化;彗星电泳试验结果显示:低、中剂量组细胞彗星头逐渐减小,彗尾较长且近似呈圆形,而高剂量组细胞彗星头较大,尾部相对较短。The essential oil from fingered citron was extracted by steam distillation to determine the cell activity by the MTT method;the inverted microscope and the Hoechst 33342/PI fluorescent staining and scanning electron microscopy were used to observe the morphological changes of apoptosis and putrescence ; DNA damage was detected by comet assay. The results showed that the volatile oil of fingered citron could obviously inhibit the cell proliferation of B16 melanoma （P 〈0. 05）,which was subject to the dose;after the treatment （6 h） of B16 melanoma with the vola- tile oil of fingered citron, it was observed under the invested microscope that the cell body of the low - medium dose group （62.5,125 and 250 I^g/mL） reduced,the morphological change was irregular,the cell debris increases and some cells grew in bulk, while the high dose group （500 μg/mL） showed morphological changes including ruptured membranes, cytoplasm spillover and increased fragments. With a fluorescence microscope, it could be seen that the low -medium dose group presented nucleus pycnosis, nuclear chromatin concentration, peripherization and other typical apoptosis characteristics,while the high dose group showed PI tinged red of nucleus, uniform distribution of nuclear chromatin and characteristics of cytoclasis;under the scanning electron microscope, it was observed that t he cellular morphology of the test group was irregular, cell surfaces were uneven, the microvillus reduced and other changes were found in cellular morphology. Moreover, the test results of the comet assay showed that in the low or middle dose groups, comet beads were smaller and comet tails were longer, compared with those in the high dose group.

关 键 词：佛手 挥发油 抑制作用 细胞凋亡 坏死 

分 类 号：R285.5[医药卫生—中药学]