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目的:研究漆姑草(HSJ)提取物对人白血病细胞株K562细胞增殖和凋亡的影响,探讨其诱导凋亡的作用机制。方法:采用不同浓度的HSJ作用K562细胞后,四甲基偶氮唑蓝(MTT)比色法检测细胞增殖抑制率;TdT酶介导的末端缺失原位标记(TUNEL)法检测细胞凋亡率;免疫细胞化学法检测Bcl-2和Bax蛋白的表达;比色法检测Caspase-3和Caspase-9的活性。结果:HSJ对K562细胞的增殖抑制率随药物剂量增大而增加;HSJ作用后细胞凋亡率增加;Bcl-2蛋白阳性表达率下降,平均透光度增加;Bax蛋白阳性表达率增加,平均透光度下降;Caspase-3和Caspase-9的活性增强,与阴性对照组比较差异具有统计学意义(P<0.05)。结论:HSJ能够诱导K562细胞凋亡;其诱导凋亡作用可能与下调Bcl-2蛋白表达水平,上调Bax蛋白表达水平,增强Caspase-3和Caspase-9的活性有关。
OBJECTIVE: To study the effect of extract of Artemisia japonica (L.) on proliferation and apoptosis of human leukemia cell line K562 and explore its mechanism of apoptosis. Methods: K562 cells were treated with different concentrations of HSJ, and MTT assay was used to detect the cell proliferation inhibition rate. TdT-mediated terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect the apoptosis rate The expressions of Bcl-2 and Bax proteins were detected by immunocytochemistry. The activities of Caspase-3 and Caspase-9 were detected by colorimetric assay. Results: The inhibitory rate of HSJ on K562 cells increased with the increase of the dose of HSJ. The apoptosis rate of HSJ increased, the expression of Bcl-2 protein decreased, the average transmittance increased, the expression of Bax protein increased, the average The light transmittance decreased; the activity of Caspase-3 and Caspase-9 increased, compared with the negative control group, the difference was statistically significant (P <0.05). CONCLUSION: HSJ can induce apoptosis in K562 cells. The apoptosis induced by HSJ may be related to the down-regulation of Bcl-2 protein expression, up-regulation of Bax protein expression and enhancement of Caspase-3 and Caspase-9 activity.