苦参碱you导A549细胞凋亡的机制研究.docVIP

　　苦参碱诱导A549细胞凋亡的机制研究 作者：范临夏　陶晓南　蔡曦光　刘华 【摘要】 目的：观察苦参碱抑制人肺腺癌细胞A549的增殖作用，并探讨其分子机制. 方法：采用四甲基偶氮唑蓝(MTT)比色法、电子显微镜、细胞周期分析法检测A549细胞凋亡；采用流式细胞术(FCM)测定细胞Bcl2, Bax及caspase3蛋白表达水平. 结果：苦参碱对A549细胞的增殖具有明显的抑制作用，并呈现剂量和时间依赖关系；电镜观察呈现典型的凋亡形态改变；细胞周期分析显示细胞增殖发生S/G2期阻滞；苦参碱能显著降低Bcl2蛋白表达(Plt;0.01)；而显著升高Bax蛋白表达水平及caspase3活性. 结论：苦参碱可显著抑制A549细胞的增殖并诱导其凋亡，其机制可能与下调Bcl2表达水平改变线粒体膜通透性有关. 【关键词】 苦参碱；肺腺癌；凋亡；Bcl2；Bax；caspase3 　　【Abstract】 AIM: To study the inhibitory effect of matrine on the proliferation of A549 lung adenocarcinoma cells and its mechanism. METHODS: MTT assay,electron microscopy, and cell cycle analysis were used to examine the apoptosis in A549 cells. Expression levels of Bcl2, Bax and caspase3 proteins were measured with flow cytometry. RESULTS: Matrine exerted antiproliferative activity on A549 cells in a dose and timedependent manner. Typical apoptotic morphological changes of A549 lung adenocarcinoma cells were observed by electron microscope. Cell cycle analysis indicated that Sphase proportion was increased, as well as S/G2 phase was apparently arrested. Bcl2 protein expression was dramatically decreased (Plt;0.01) whereas the expression of Bax and caspase3 was increased in A549 cells after treated by matrine. CONCLUSION: Matrine can exert the significant growthinhibitory effect on A549 cells and induce the cell apoptosis, which is probably related with the downregulation of Bcl2 expression via changing the permeability of mitochondrial membrane. 　　【Keywords】 matrine; lung adenocarcinoma; apoptosis; Bcl2; Bax; caspase3 　　0引言 　　苦参碱作为中药苦参的主要活性成分，临床主要用于镇痛、抑菌、抗病毒以及防治肝纤维化. 近年研究显示，苦参碱具有明显的抗肿瘤作用［1-2］. 雷怀定等［3］研究发现苦参碱对体外人肺癌A549细胞具有增殖抑制作用，我们观察苦参碱诱导人肺癌A549细胞凋亡及其机制，为苦参碱用于临床抗肿瘤提供理论依据. 　　1材料和方法 　　1.1材料 　　苦参碱购自宁夏盐池，配制为20 g/L的无菌溶液备用. MTT购自Sigma公司. 流式细胞术（flow cytometry, FCM）用mAb（FITCconjugated mouse antihuman Bax monoclonal antibody and IgG1 isotype control, FITCconjugated American Hamster antihuman Bcl2 monoclonal and American Hamster monoclonal IgG isotype control）为美国Calta