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目的研究蝙蝠葛活性成分体外对人白血病K562细胞株的抑制增殖和诱导凋亡作用。方法应用MTT法检测蝙蝠葛活性成分对体外培养的人白血病K562细胞株的抑制增殖作用的影响及细胞毒活性;通过倒置显微镜对细胞进行形态学观察;通过免疫细胞化学S-P法测定凋亡关键效应酶Caspase-9、Caspase-8和Caspase-3的表达情况。结果(1)MTT比色法结果表明,蝙蝠葛活性成分对人白血病K562细胞株的增殖抑制作用呈剂量依赖性(P<0.01);(2)倒置显微镜观察:蝙蝠葛活性成分20μg/ml作用后早期细胞出现凋亡形态学变化:细胞膜鼓泡、凋亡小体形成等;作用晚期细胞发生膜破裂坏死;(3)免疫细胞化学结果表明:蝙蝠葛活性成分作用后加药组Caspase-9、Caspase-8和Caspase-3表达增加,与对照组相比较均有显著差异(P<0.01)。结论蝙蝠葛活性成分对人白血病K562细胞株具有抑制增殖和诱导凋亡作用。
Objective To study the inhibitory effect of the active ingredients of bats on proliferation and apoptosis of human leukemia K562 cell line in vitro. Methods MTT assay was used to determine the inhibitory effect and the cytotoxic activity of the active ingredients of Douthegrafenum on human leukemia K562 cells cultured in vitro. The morphological changes of the cells were observed by inverted microscope. The key apoptosis effects were determined by immunocytochemistry Caspase-9, Caspase-8 and Caspase-3 expression. Results (1) The results of MTT assay showed that the active components of Daphoot against proliferation of human leukemia K562 cells in a dose-dependent manner (P <0.01); (2) Inverted microscope observation: (3) The results of immunocytochemistry showed that the activity of Caspase-9 in the dosing group increased significantly (P <0.05), and the apoptotic morphological changes in the early post- , Caspase-8 and Caspase-3 expression increased, compared with the control group were significantly different (P <0.01). Conclusions The active ingredients from Daphne vaporariorum inhibit proliferation and induce apoptosis of human leukemia K562 cell line.