川芎嗪对白血病细胞凋亡及凋亡相关蛋白表达的影响

Effects of tetramethylpyrazine on apoptosis of human leukemia cells and the expressions of apoptoticrelevant proteins

目的 :探讨川芎嗪注射液对白血病细胞凋亡及凋亡相关蛋白表达的影响。方法 :应用不同质量浓度(0.5、1.0、1.5、2.0和2.5 mg/mL)的川芎嗪溶液作用人白血病U937细胞24、48和72 h后,采用CCK-8法检测川芎嗪对U937细胞的增殖抑制率;FCM法检测川芎嗪对U937细胞凋亡及细胞周期分布的影响;蛋白质印迹法检测细胞内凋亡相关蛋白survivin、Bcl-2、Bax和caspase-3的表达情况。结果 :川芎嗪能明显抑制U937细胞增殖(P<0.05),并呈浓度和时间依赖性(P<0.05)。经不同质量浓度(0.5、1.0和1.5 mg/mL)的川芎嗪溶液处理24 h后,U937细胞的S期细胞比例均增加(P<0.05),G2/M期细胞比例均减少(P<0.05),细胞凋亡率明显升高(P<0.05),并呈浓度依赖性。川芎嗪作用能明显降低survivin和Bcl-2蛋白的表达水平(P<0.05),提高Bax和caspase-3蛋白表达水平(P<0.05),并呈浓度依赖性。结论 :川芎嗪可能通过诱导U937细胞凋亡发挥抗肿瘤作用,其机制可能与下调survivin和Bcl-2蛋白表达,并上调Bax和caspase-3蛋白表达有关。

Objective： To explore the effects of tetramethylpyrazine injection on apoptosis of human leukemia cells and the expressions of apoptotic-relevant proteins. Methods： Leukemia cell line U937 cells were treated with different concentrations（0.5, 1.0, 1.5, 2.0 and 2.5 mg/mL） of tetramethylpyrazine for 24, 48 and 72 h. Then the proliferation inhibitory rate of U937 cells was detected by CCK-8（cell counting kit-8） method. The cell cycle distribution and the apoptosis of U937 cells were detected by fl ow cytometry. The expressions of apoptotic-relevant proteins survivin, Bcl-2, Bax and caspase-3 in U937 cells were measured by Western blotting. Results： Tetramethylpyrazine significantly inhibited the cell growth of U937 cells in a time- and dose-dependent manner（P 〈 0.05）. After treatment with different concentrations（0.5, 1.0, 1.5 mg/mL） of tetramethylpyrazine for 24 h, the proportion of U937 cells in S phase increased（P 〈 0.05）, but which in G2/M phase decreased（P 〈 0.05）, as well as the apoptosis rate increased signifi cantly in a dose-dependent fashion（P 〈 0.05）. The expression levels of survivin and Bcl-2 proteins were reduced（P 〈 0.05）, and the expression levels of Bax and caspase-3 proteins were elevated（P 〈 0.05） in a dose-dependent fashion in U937 cells after tetramethylpyrazine treatment. Conclusion： Tetramethylpyrazine injection can signifi cantly induce apoptosis of U937 cells resulting in antineoplastic effect. Its mechamism may be related to down-regulation of survivin and Bcl-2 protein expressions and up-regulation of Bax and caspase-3 protein expressions.