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目的:检测不同浓度的丹参酮I、丹参酮IIa、隐丹参酮对小鼠黑色素瘤B16细胞增殖活力的影响;测定丹参酮I、丹参酮IIa、隐丹参酮对B16细胞周期的影响。方法:采用甲基噻唑基四唑比色法(MTT法)及显微镜细胞观察法检测丹参酮I、丹参酮IIa、隐丹参酮对小鼠黑色素瘤B16细胞增殖的抑制作用。通过流式细胞仪检测丹参三种成分对B16细胞周期的影响。结果:不同浓度(2.5μg/m L、5μg/m L、10μg/m L、20μg/m L、30μg/m L)的丹参酮I、丹参酮IIa和隐丹参酮均能显著抑制黑色素瘤细胞B16的增殖活性,且呈明显的剂量效应和时间效应。丹参酮I的抑瘤效果最显著,其72 h的IC50仅为0.25μg/m L。高浓度(30μg/m L)的丹参酮I、丹参酮IIa和隐丹参酮均可在作用24 h后抑制小鼠黑色素瘤B16细胞的有丝分裂,主要阻滞于G1期,但其作用效果不显著。结论:不同浓度(2.5μg/m L、5μg/m L、10μg/m L、20μg/m L、30μg/m L)的丹参酮I、丹参酮IIa和隐丹参酮均能显著抑制黑色素瘤细胞B16的增殖活性并阻滞其细胞周期,且具有剂量依赖性。高浓度(30μg/m L)的丹参酮I和丹参酮IIa均可显著引起小鼠黑色素瘤B16细胞的凋亡,其中丹参酮I效果最显著。
OBJECTIVE: To detect the effects of tanshinone I, tanshinone IIa and cryptotanshinone on the proliferation of mouse melanoma B16 cells. The effects of tanshinone I, tanshinone IIa and cryptotanshinone on the cell cycle of B16 were determined. Methods: The inhibitory effects of tanshinone I, tanshinone IIa and cryptotanshinone on the proliferation of mouse melanoma B16 cells were detected by methyl thiazolyl tetrazolium (MTT) assay and microscope observation. The effects of three components of Salvia miltiorrhiza on the cell cycle of B16 were detected by flow cytometry. Results: Tanshinone I, tanshinone IIa and cryptotanshinone with different concentrations (2.5μg / ml, 5μg / ml, 10μg / ml, 20μg / ml and 30μg / ml) could significantly inhibit the proliferation of B16 melanoma cells Activity, and showed obvious dose effect and time effect. Tanshinone I had the most significant antitumor effect with an IC50 of only 0.25 μg / m L at 72 h. Tanshinone I, tanshinone IIa and cryptotanshinone at high concentration (30μg / m L) all inhibited the mitosis of mouse melanoma B16 cells 24 h after treatment, and mainly blocked in G1 phase, but the effect was insignificant. CONCLUSION: Tanshinone I, tanshinone IIa and cryptotanshinone at different concentrations (2.5μg / ml, 5μg / ml, 10μg / ml, 20μg / ml and 30μg / ml respectively) can significantly inhibit the proliferation of B16 melanoma cells Activity and block its cell cycle, and in a dose-dependent manner. Tanshinone I and tanshinone IIa at high concentration (30μg / ml) could significantly induce the apoptosis of mouse melanoma B16 cells, among which tanshinone I was the most effective.