苦参碱对唾液腺ACC-M细胞凋亡及Bcl-2、Bax表达的影响

The influence of Matrine on apoptosis and Bcl-2/Bax expression of salivary ACC-M cells

目的:探讨中药提取物苦参碱(Matrine)对人唾液腺腺样囊性癌(SACC)细胞系ACC-M的凋亡诱导作用及其发生的机制。方法:体外培养ACC-M细胞,用不同浓度的苦参碱对体外培养的ACC-M细胞进行干预。分别采用MTT、免疫细胞化学染色、间接免疫荧光联合流式细胞仪定量检测等方法,观察其对ACC-M细胞的增殖抑制和凋亡诱导作用;并对凋亡相关Bcl-2与Bax蛋白表达的变化进行定性与定量检测。计量资料采用x±s表示,采用SPSS10.0统计软件包对数据进行t检验。结果:经不同浓度的苦参碱处理后的ACC-M细胞,其生长受到明显的抑制,细胞凋亡指数随药物浓度增加而明显增加(从4.21%到27.47%);0.75mg/ml的苦参碱作用48h后,其Bcl-2表达明显下降。定量结果显示,平均荧光强度从570.6降至512.4;而Bax表达则明显上升,定量结果显示,平均荧光强度从197.5升至313.7,P<0.05,有显著性差异。结论:一定浓度的苦参碱能抑制ACC-M细胞的增殖,促进其凋亡;其机制可能与调控Bcl-2、Bax表达有关。

PURPOSE： To investigate the effects of Matrine that is active component of Chinese traditional medicine on inducing apoptosis and explore the mechanism of action in human adenoid cystic carcinoma cell line. METHODS： Different concentrations of Matrine were used in the culture of ACC-M cells. Proliferation and apoptosis of ACC-M cells were observed by ways of MTT, immunocytochemistry, indirect immunofluorescene and flow cytometry quantitative analysis,etc. At the same time, the expression of Bcl-2 and Bax protein was determined by qualitative and quantitative analysis. The data were analyzed by Student＇s t test using SPSS10.0 software package. RESULTS： Proliferation of ACC- M cells was inhibited effectively after being treated by different concentrations of Matrine. The rate of apoptosis was increasing with increased concentration of Matrine. The expression of Bcl-2 was decreasing in the experimental groups by two ways. Otherwise, the expression of Bax protein was significantly higher than the control group. Quantitative analysis showed that the average fluorescen intensity of Bcl-2 was falling from 570.6±11.62 to 512.4±5.76 between the control group and the experimental group. But the average fluorescen intensity of Bax was increasing from 197.5±6.92 to 313.7± 6.88 between the control group and the experimental group. The difference was significant （P〈0.05）. CONCLUSIONS： Matrine can inhibit proliferation and promote apoptosis of ACC-M cells effectively, the mechanism may be correlated to the up-regnlation of Bax and down-regnlation of Bcl-2.