荆芥挥发油抗肿瘤作用的研究

Research on Anti-tumor Effect of Essential Oil in Schizonepeta Tenuifo-lia Briq and Its Inducing Apoptosis

目的:研究荆芥挥发油的抗肿瘤活性,确定其最小杀伤肿瘤细胞的剂量;同时观察诱导细胞凋亡的最小剂量以确定其抗肿瘤机制。方法:根据药物作用的不同时间,分为24h、48h、72h三组,每组分设不同剂量(0.125mg/ml-16mg/ml)。以吐温-80作为助溶剂对荆芥挥发油倍比稀释,对人A549肺癌细胞进行抗肿瘤试验,采用SRB方法和形态学确认细胞杀伤率,并采用荧光染料Hochest33258染色检测细胞的凋亡水平。结果:大于或等于4mg/ml以上剂量的荆芥挥发油提取物对A549具有较高的抑瘤率,最低可达91.2%;最低抑瘤浓度为1mg/ml,抑瘤率为43.3%。形态学观察结果表明,被杀伤的细胞体积萎缩、细胞膜皱缩、成活细胞数稀疏、数量明显低于空白对照组。不同给药时间的研究结果表明,与作用时间为24h组比较,48h及72h组的杀伤作用效果更佳(P<0.05或P<0.01)。以低于杀伤浓度的剂量(0.125-1mg/ml)作用于A54972h,利用Hochest33258观察其对A549细胞凋亡的影响,结果表明,0.25mg/ml-1mg/ml剂量的荆芥挥发油提取物作用于细胞72h对肿瘤细胞具有明显的诱导细胞凋亡的作用。结论:高浓度(4mg/ml-16mg/ml)的荆芥挥发油对人肺癌A549细胞株有杀伤作用,低浓度(0.25mg/ml-1mg/ml)的荆芥挥发油对A549有诱导细胞凋亡的作用。

Objective： To extract the essential oil from Schizonepeta tenuifo- ha Briq for making experiments of anti human lung cancer and inducing its apeptosis so as to observe the anti - tumor activity, the minimum dose of killing tumor cells and inducing cells apoptosis of the essential oil as well as its mechanism. Methods： The improved steam distillated method was used to isolated the essential oil of Schizonepeta tenuifo - ha Briq. Tween - 80 was used to dissolve the essential oil and diluted by saline. The human lung cancer cell line A549 was applied in this study. The anti - tumor effect was assayed by the SRB method and the inducing- apeptosis as assayed by hochest33258 stain. The experiment groups were divided into the time-dependence group （24h, 48h, 72h） and the dose-dependence group （0.25 mg/ml - 16 mg/ml）. Results： 4 mg/ ml or more of essential oil in Schizonepeta tenuifo - ha Briq obviously inhibited the growth of A549 with the highest inhibitive rate being 91.2% while the minimum inhibitive dose was 1 mg/ml with the rate being 43.3%. According to the morphological observation, the atrophy of necrotic cells and the number of the surviving cells in the experiment groups were lower than in the control groups. More over, the best effect were achieved at the time of 48 h and 72 h, and obvious effect on inducing ceil apeptosis of oil were found for dose between 0.25 mg/ml and 1 mg/ml in 72h. Conclusions： The high dose （4 mg/ml - 16 mg/ml） of essential oil in Schizonepeta tenuifo - lia Briq can kill the cells of human lung cancer A549, and the low dose can inducing its apoptosis.