大蒜素诱导LM-8细胞凋亡及相关基因表达的实验研究

Apoptosis of LM-8 Cells Induced by Allicin

目的:探讨大蒜素对C3H小鼠骨肉瘤细胞株LM-8细胞增殖的影响及与bcl-2、bax凋亡基因表达的关系,以初步探索其诱导凋亡的机制。方法:用水溶性四唑盐WST-8比色法检测大蒜素对LM-8细胞株生长的影响;倒置显微镜观察细胞形态变化;流式细胞仪检测细胞调亡率的变化;免疫细胞组化技术检测bcl-2和bax基因表达。结果:大蒜素能明显抑制LM-8细胞的生长,与大蒜素的浓度和时间有关,在72h其药物半数抑制率浓度是11.09μg/ml;5μg/ml、10μg/ml和15μg/ml浓度的大蒜素作用72h后可诱导LM-8细胞凋亡(P<0.05);大蒜素可以降低bcl-2表达,增强bax表达,经15μg/ml大蒜素作用72h,bcl-2和bax基因表达阳性率及bcl-2/bax分别为(7.24±1.18)%(、47.56±7.43)%和0.15±0.02,对照组分别为(24.40±9.96)%、(11.28±1.31)%和2.18±0.93,与对照组比较,差异有显著性意义(P<0.05)。结论:大蒜素可以诱导LM-8细胞凋亡,其机制涉及到bcl-2表达上调及bax表达下调。

Objective： To investigate the effects of Allicin on LM-8 cells and probe the possible mechanism. Method：Water -soluble tetrazolium salt （WST-8） assay was used to detect the growth inhibitory effect in 24, 48 and 72 hours, respectively. Light microscopy was performed to observe the morphologic changes of cells. Flow cytometry was used to evaluate apoptosis. The gene expression of bcl-2 and bax was examined by immunohistochemistry. Result： Allicin inhibited the growth of LM-8, and the inhibitory rate of cells treated by Allicin （11.09μg/ml） for 72 hours was 50%. Cells cultured with Allicin were observed to present typical apoptosis changes. Flow cytometry revealed a dose-dependent relationship. The positive protein expression rates of bcl-2 and bax were （7.24＋ 1.18）% and （47.56±7.43）% respectively with Allicin of 15tlg/ml in 72 hours, while the controls being （24.40±9.96）% and （ 11.28±1.31 ）% （ p〈0.05）, and the value of Bcl- 2/ Bax was related with the dose of Alllcin（p〈0.05）. Conclusion： Allicin can induce apoptosis in LM-8 cells by the downregulation of bcl-2 expression and upregulation of bax expression.