茅莓总皂苷对黑色素瘤的抗肿瘤作用研究

目的：评价茅莓总皂苷对黑色素瘤的体内、外抗肿瘤药效作用。方法：A375人恶性黑色素瘤细胞体外常规传代培养,随机分5组,分别加入环磷酰胺,茅莓总皂苷高、中、低(0.01,0.001,0.000 1 mg·mL^-1)3个质量浓度,以不加药作阴性对照,MTT法检测茅莓总皂苷的体外抗肿瘤作用；用B16黑色素瘤细胞移植小鼠,随机分阳性药(环磷酰胺)组,茅莓总皂苷高、中、低(100,50,25 mg·kg^-1)剂量组及阴性药(生理盐水)组,观察体内抑瘤效果、荷瘤小鼠生存时间和生命延长率；同时采用TUNEL法检测茅莓总皂苷对B16黑色素瘤细胞凋亡的影响。结果：体外抑瘤试验显示,高、中质量浓度茅莓总皂苷组与阴性对照组比较,P<0.05,有统计学意义；体内抑瘤试验显示,高剂量组平均瘤体积明显低于阴性对照组(P<0.05),抑瘤率37.02%；荷B16黑色素瘤小鼠用茅莓总皂苷治疗后存活时间增加；茅莓总皂苷高、中、低剂量组黑色素瘤细胞凋亡指数分别为32.5%,20.5%和5.5%,与阴性对照组(3.5%)比较差异有统计学意义。结论：茅莓总皂苷对体内、外黑色素瘤细胞有抑制作用,并呈量效依赖关系,能通过促进黑色素瘤细胞凋亡而发挥抗肿瘤活性。

Antitumour effect of total saponins of Rubus parvifolius on malignant melanoma

Objective: To evaluate the antitumor effect of total saponins of R. parvifolius on malignant melanoma. Method: The human malignant melanoma A375 cells were regularlly subcultured in vitro, and were divided into five groups contained positive control group (CTX), high concentration (0.01 mg·mL^-1) and middle concentration (0.001 mg·mL^-1) and low concentration (0.000 1 mg·mL^-1) total saponins of R. parvifolius groups and negative control group. By using MTT colorimetric method, the cell viability was measured. B16 melanoma cells were transplanted to mice, which were divided into positive control group,high dose (100 mg·kg^-1) and middle dose (50 mg·kg^-1) and low dose (25 mg·kg^-1) total saponins of R. parvifolius groups and negative control group. The inhibition effect of the tumor in vivo, mean survival time and rate of life-elongation of the mice were observed. TUNEL method was used to detect the apoptosis of B16 malignant melanoma. Result: Antitumor assay in vitro showed that the absorbency increased in the concentration of 0.01, 0.001 mg·mL^-1 with statistical significance (P<0.05 vs negative contro1). Antitumor assay in vivo showed that the tumor inhibitory rate of high dose (100 mg·kg^-1) and middle dose (50 mg·kg^-1) of total saponins of R. parvifolius were 37.02% and 30.61%, respectively. Loaded tumor mouse survival duration could be prolonged. The apoptosis indexes of B16 tumor cells in three treatment groups were 32.5%, 20.5% and 5.5%, respective and there was statistical significance (P<0.05 vs negative contro1). Conclusion: The total saponins of R. parvifolius has remarkable inhibition of proliferation of malignant melanoma cells in vivo and in vitro and exerts antitumor activities through promoting tumor cell apoptosis.