加味四君子汤含药血清对B16恶性黑色素瘤细胞增殖和凋亡的影响

Effect of Jiawei Sijunzi Decoction Containing Serum on B16 Malignant Melanoma Cell Proliferation and Apoptosis

目的:研究加味四君子汤含药血清对B16恶性黑色素瘤细胞增殖和凋亡的影响。方法:应用血清药理学方法给药,37℃5%CO2的恒温培养箱中培养B16恶性黑色素瘤细胞,收集对数期细胞,终浓度10%的小鼠含药血清,分别作用12,24,48 h。实验分为空白对照组、血清对照组、加味四君子汤含药血清组(高、中、低3个剂量组)、顺铂组,每组含5个样本数。分别应用MTT法、流式细胞仪Annexin V-FITC/PI法检测加味四君子汤含药血清对B16恶性黑色素瘤细胞增殖和凋亡的影响。结果:加味四君子汤含药血清作用12,24,48 h,与同时间段空白对照组和血清对照组相比,以剂量依赖方式抑制B16恶性黑色素细胞增殖,中剂量加味四君子汤含药血清组24,48 h抑制率为50.22%,53.81%,且48 h与24 h相比没有显著性变化;与空白对照组和血清对照组相比,中、高剂量加味四君子汤含药血清组、顺铂组的凋亡率分别为(19.70±0.98)%,(25.21±1.03)%,(21.31±1.00)%,显著促进B16恶性黑色素瘤细胞的凋亡(P<0.05),但顺铂组凋亡率高于中剂量中药组,低于高剂量中药组。结论:加味四君子汤含药血清抑制B16恶性黑色素瘤细胞增殖;促进B16恶性黑色素瘤细胞的凋亡,这可能是加味四君子汤抗恶性黑色素瘤的机制之一。

Objective： To investigate effect of Jiawei Sijunzi decoction containing serum on B16 malignant melanoma cell proliferation and apoptosis. Method： Jiawei Sijunzi decoction was administrated by serum pharmacological method, which B16 melanoma cells were cultured in 37 ℃ and 5% CO2 thermostat incubator, and logarithmic phase cells were collected and exposed to final concentration 10% mice containing serum in 12, 24,48 h, respectively.The experimental group were divided into blank control group, serum control group, traditional Chinese medicine group （high dose,middle dose and low dose）, and cisplatin group, and each group contained 5 samples. MTT and Flow cytometry Annexin V-FITC/PI assay were used to analyse the Jiawei Sijunzi decoction containing serum influence on B16 malignant melanoma cell proliferation and apoptosis respectively. Result： A time-dependent increase in inhibiting B16 malignant melanoma cell proliferation by Jiawei Sijunzi decoction containing serum was observed compared to blank control group, serum control group in the same time period, and middle dose traditional Chinese medicine group inhibition rate was 50.22% in 12 h and 53.81% in 24 h respectively,and reached the half of inhibition rate, and half of inhibition rate in 48 hours period and 24 hours period were compared to no significant change. Middle dose traditional Chinese medicine group, high dose traditional Chinese medicine group and cisplatin group （5 mgL-1） apoptosis rate were （19.70±0.98）%,（25.21±1.03）%,（21.31±1.00）% respectively, and promote significantly B16 malignant melanoma cell apoptosis compared to the blank control group and serum control group, the rate of cisplatin group apoptosis is high than middle dose traditional Chinese medicine group, and lower than high dose traditional Chinese medicine group. Conclusion： Jiawei Sijunzi decoction containing serum can inhibit the proliferation of B16 malignant melanoma cells, and promote the apoptosis of B16 malignant melanoma cells, which could be the therepy mechanism of Jiawei Sijunzi decoction antitumor.