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目的:探讨传统藏药总状绿绒蒿体外对人慢性髓系白血病细胞株K562细胞增殖活性的影响,并阐明其相关作用机制。方法:不同浓度(30、60、90、120和150mg.L-1)总状绿绒蒿醇提物作用于体外培养的白血病K562细胞,同时设置空白对照组,MTT法检测各组细胞增殖抑制率,倒置显微镜观察细胞形态学改变,单细胞凝胶电泳检测细胞DNA损伤,流式细胞术检测细胞周期时相变化。结果:作用于K562细胞24、48和72h后,不同浓度总状绿绒蒿醇提物组K562细胞生长受到明显抑制,与空白对照组比较,其增殖抑制率明显增加(P<0.05),并呈浓度和时间依赖性。倒置显微镜下观察,随着总状绿绒蒿醇提物浓度增加,K562细胞数量减少,细胞形态不规则,轮廓模糊,细胞碎片增多。单细胞凝胶电泳(SCGE)检测,与空白对照组比较,60、90和120mg.L-1总状绿绒蒿醇提物组K562细核内DNA片段化、TailDNA%升高,差异有统计学意义(P<0.05或P<0.01)。细胞周期检测,60、90和120mg.L-1总状绿绒蒿醇提物作用于K562细胞24h后,G0/G1期、S期细胞所占比例逐渐下降,G2/M期细胞所占比例逐渐升高,分别为(1.88±0.30)%、(5.46±0.57)%和(19.80±1.03)%,各浓度组与空白对照组[(1.10±0.12)%]比较差异均有统计学意义(P<0.05),总状绿绒蒿醇提物组K562细胞周期发生了G2/M期阻滞。结论:总状绿绒蒿醇提物对K562细胞有显著的增殖抑制作用,其机制可能与诱导DNA损伤引发G2/M期阻滞有关联。
OBJECTIVE: To investigate the effect of the traditional Tibetan Medicinal Materia Medica on the proliferation of human chronic myeloid leukemia K562 cells in vitro and to elucidate its related mechanism. Methods: Alcohol extract of different concentrations (30, 60, 90, 120 and 150 mg.L-1) of Procyon mulchetum on K562 cells were cultured in vitro. At the same time, a blank control group was set up. MTT assay was used to detect the cell proliferation inhibition The morphological changes of cells were observed with inverted microscope, DNA damage was detected by single cell gel electrophoresis, and the cell cycle phase changes were detected by flow cytometry. Results: After treated with K562 cells at different concentrations for 24, 48 and 72 hours, the growth of K562 cells was significantly inhibited. Compared with the blank control group, the proliferation inhibition rate was significantly increased (P <0.05) Concentration and time-dependent. Under inverted microscope, with the increase of the concentration of alcohol extract of Meconopsis pumila, the number of K562 cells decreased, the cell shape was irregular, the outline was blurred and the number of cell debris increased. Single cell gel electrophoresis (SCGE) detection, compared with the blank control group, 60,90 and 120mg.L-1 ethanol extract of Prochlorococcus argenchei K562 nuclear DNA fragmentation, TailDNA% increased, the difference was statistically Significance (P <0.05 or P <0.01). Cell cycle detection, 60,90 and 120mg.L-1 Prochloromycoli ethanol extract on K562 cells 24h, G0 / G1 phase, S phase cells decreased gradually, G2 / M phase cells proportion (1.88 ± 0.30)%, (5.46 ± 0.57)% and (19.80 ± 1.03)%, respectively. There was significant difference between the control group and the control group [(1.10 ± 0.12)%] ( P <0.05). G2 / M arrest occurred in the cell cycle of K562 cells. CONCLUSION: Alcohol extract of Meconopsis litura has a significant inhibitory effect on K562 cells. The mechanism may be related to the induction of G2 / M arrest induced by DNA damage.