佛手挥发油对B16细胞增殖及其酪氨酸酶活性的影响  

Effects of fingered citron essential oil on the proliferation and tyrosinase activity of B16 melanoma cells in vitro

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

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004

出　　处：《浙江师范大学学报（自然科学版）》2011年第2期202-206,共5页Journal of Zhejiang Normal University:Natural Sciences

摘　　要：为检测佛手挥发油对B16黑色素瘤细胞增殖及其酪氨酸酶活性的影响,采用台盼蓝排斥法测定了细胞存活率、瑞-姬氏混染法观察了细胞形态变化、吖啶橙/溴化乙锭染色法检测了细胞的凋亡与坏死,用酶学方法测定了细胞中酪氨酸酶活性.结果表明:佛手挥发油对B16黑色素瘤细胞增殖具有显著的抑制作用,呈剂量依赖性;62.50,125.00和250.00μg.mL-1佛手挥发油处理细胞6 h后,细胞核染色质凝聚于核膜内侧,呈固缩状或圆珠状,表现为典型的细胞凋亡特征;500.00μg.mL-1佛手挥发油处理细胞后,细胞膜破裂,出现细胞碎片,说明细胞已经坏死.酪氨酸酶活性检测结果显示:佛手挥发油对B16黑色素瘤细胞酪氨酸酶活性具有明显的抑制作用,且呈剂量依赖性.To investigate the effects of fingered citron essential oil on proliferation of B16 melanoma cells and tyrosinase activity in vitro,typan blue exclusion method was used to test the survival rate of B16 melanoma cells.The morphological changes of apoptosis or necrosis in B16 cells were observed by inverted microscope and fluorescence microscope.Tyrosinase activity was tested by using enzyme-kinetic-method.The experiment results showed that,compared with the control group,62.50~500.00 μg·mL-1 fingered citron essential oil had obvious inhibition on the proliferation of B16 cell and this effect was dose-dependent.Moreover,the cells in 62.50,125.00 and 250.00 μg·mL-1 groups exhibited typical characteristics of apoptosis in morphology such as chromatin margination and pyknosis.In contrast,the cells in 500.00 μg·mL-1 group showed characteristics of necrotic including cytomembrane rupture and cell debris appear.Furtermore,the results from tyrosinase activity test indicated that fingered citron essential oil could significantly inhibit the tyrosinase activity,and this action appeared a dose-dependent effect relation.

关 键 词：佛手挥发油 B16黑色素瘤细胞 细胞增殖 细胞凋亡 酪氨酸酶活性 

分 类 号：R285.5[医药卫生—中药学]