牛蒡子苷元对小鼠红白血病FBL-3细胞凋亡的影响及相关机制

Effect of arctigenin on apoptosis of mouse erythroleukemia FBL-3 cells and its mechanism

目的探讨牛蒡子苷元对小鼠红白血病细胞株FBL-3细胞凋亡的影响及机制。方法以小鼠红白血病FBL-3细胞为研究对象，以不加药的FBL-3细胞为对照组，20mg／L牛蒡子苷元处理24h的FBL-3细胞为实验组。通过琼脂糖凝胶电泳和流式细胞术观察牛蒡子苷元对小鼠红白血病FBL一3细胞凋亡的影响，通过反转录-聚合酶链反应（RT-PCR）法检测牛蒡子苷元处理前后FBL-3细胞凋亡相关基因表达的变化。结果电泳结果显示，与对照组相比，实验组可见明显的DNA“梯子”样断裂变化。流式细胞仪结果显示，与对照组相比，实验组细胞凋亡数量明显增加（t=60．681，P=0．000）。RT—PCR结果显示，与对照组相比，实验组细胞抗凋亡基因Bcl-2和lAP-J表达显著下调（t=14．732、29．702，P均=0．000），而Bcl—XL表达没有明显变化（t=0．209，P=0．844）；促凋亡基因Box和Smac表达明显上调（t=6．721、8．499，P=0．003、0．001）。结论牛蒡子苷元可诱导FBL-3细胞凋亡，其机制可能与抗凋亡基因Bcl-2租IAP-J表达下调及促凋亡基因bax和Smac表达上调有关。

Objective To investigate the effect of arctigenin on the apoptosis of FBL-3 cells and its mecha- nisms. Methods The mouse erythroleukemia FBL-3 cells were taken as subjects. The untreated FBL-3 parental cells were taken as the control group, and the FBL-3 cells treated with 20 mg/L arctigenin for 24 h were taken as the experi- mental group. The effects of arctigenin on the apoptosis of the mouse erythroleukemia FBL-3 cells were determined by agarose gel electrophoresis and flow cytometry assay. The changes of apoptosis-related genes were analyzed by reverse transcriptase-polymerase chain reaction（RT-PCR）. Results The agarose gel electrophoresis results revealed that the ＂ DNA ladder＂ was displayed in the experimental group compared to the control group. The flow cytometry data demon- strated that the apoptosis number of FBL-3 cells in the experimental group was markedly increased compared to that in the control group （t = 60. 681, P = 0. 000）. The RT-PCR assay results suggested that the expressions of Bcl-2 and IAP-1 gene were down-regulated in the experimental group compared to the control group （t = 14. 732,29. 702, all P = 0. 000）, while the expressions of Bax and Smac gene were up-regulated（ t = 6. 721,8. 499 ;P = 0. 003,0. 001 ） , but the expression of Bcl-XL did not change （ t = 0. 209, P = 0. 844 ）. Conclusions Aretigenin can induce the apoptosis of the mouse erythrolenkemia FBL-3 cells. Apoptosis of the mouse erythrolenkemia FBL-3 cells induced by arctigenin may be correlated to the down-regulation of Bcl-2 and IAP-1 and up-regulation of Bax as well as Smac.