X连锁凋亡抑制蛋白通路在白花蛇舌草的有效成分2-羟基-3-甲基蒽醌诱导的人白血病细胞HL-60凋亡中的作用

Roles of X-linked inhibitor of apoptosis proteins in the apoptosis of human HL-60cells induced by 2-hydroxy-3-methylanthraquinone from Hedyotis diffusa Willd

目的探讨白花蛇舌草的有效成分2-羟基-3-甲基蒽醌诱导人白血病细胞HL-60细胞凋亡的机制。方法以HL-60细胞作为体外细胞模型,当作用时间分别为0、24、48及72 h时,用Hoechst 33258荧光染色技术、Annexin V FITC/PI双染流式细胞仪检测HL-60细胞凋亡率;Western blot检测Survivin、Smac、c IAP1/2、XIAP等蛋白的表达。结果作用24 h时,即可引起HL-60细胞发生凋亡,HL-60细胞凋亡百分率随作用时间的延长而增加。Hoechst 33258荧光染色及流式细胞仪两种方法所得结果一致。随着作用时间的延长,XIAP和c IAP1/2表达逐渐下降,而Smac表达逐渐增加。结论白花蛇舌草的有效成分2-羟基-3-甲基蒽醌能够诱导HL-60细胞凋亡,XIAP信号通路在此过程中起到重要作用。

Objective To investigate the relationship between 2-hydroxy-3-methylanthraquinone from Hedyotis diffusa Willd and the apoptosis of HL-60. Methods HL-60 cell apoptosis induced by 2-hydroxy-3-methylanthraquinone from Hedyotis diffusa Willd in 0,24,48 and 72 h respectively was detected with Hochest 33258 fluorochrome staining and Annexin V FITC/PI assay. Western blot was performed to detect the expressions of Survivin, Smac, XIAP and cIAP1/2. Results The apoptotic cells were found in all the groups,and the percentage of apoptotic cells was raised in a time-dependent manner. With the time extension,there was down-regulation of XIAP and cIAP1/2,and up-regulation of Smac. However,the expression of Survivin did not change. Conclusion 2-hydroxy-3-methylanthraquinone is an ef-fective apoptosis-inducing natural compound of HL-60 cells,which suggests that 2-hydroxy-3-methylanthraquinone may have a role in future therapies for human leukemia.