基于AFM对蝙蝠葛碱诱导NB4细胞凋亡的研究  

Atomic Force Microscope-related Study on Dauricine Induced Apoptosis of NB4 Cell

作　　者：陈伟[1] 王穗湘[2] 胡小毛[3] 邢晓波[1] 蔡继业[1] 

机构地区：[1]暨南大学化学系,广东广州510632 [2]暨南大学医学院,广东广州510632 [3]暨南大学第一临床医学院,广东广州510632

出　　处：《中山大学学报（医学科学版）》2012年第4期421-428,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(30872404);"973计划"项目(2010CB/833603);中国博士后基金(20090450099;201003359)

摘　　要：【目的】探讨蝙蝠葛碱对急性早幼粒细胞白血病(APL)NB4细胞增殖的影响和可能的细胞凋亡机制。【方法】采用CCK-8法检测NB4细胞的存活率,原子力显微镜(AFM)探测蝙蝠葛碱作用NB4细胞前后细胞形态及超微结构的变化,流式细胞术检测细胞凋亡率、线粒体膜电位和活性氧的变化。【结果】蝙蝠葛碱能显著抑制NB4细胞生长增殖,20～80μmol/L蝙蝠葛碱处理细胞24 h后,细胞存活率从(86.5±11.7)%下降到(4.1±0.5)%;以10～40μmol/L蝙蝠葛碱处理NB4细胞24 h,流式细胞术分析显示,细胞凋亡率和细胞内活性氧含量上升、线粒体膜电位下降。AFM探测表明,正常NB4细胞呈圆形,细胞饱满,表面较光滑。蝙蝠葛碱处理组细胞皱缩,细胞表面的平均粗糙度增大。【结论】蝙蝠葛碱能显著抑制NB4细胞增殖并诱导其凋亡。[Objective] To investigate the role of acute promyelocytic leukemia NB4 cells in cell growth and the possible mechanism of cell apoptosis by dauricine. [Methods] In this study, CCK-8 assay was used to detect the proliferation rate of NB4 cell. atomic force microscope （AFM） was used to detect the changes in morphological and ultrastructure of NB4 cells before and after dauricine treatment. Cell apoptosis rate, mitochondrial membrane potential （MMP）, and reactive oxygen species （ROS） were detected by flow cytometry. [Results] The result showed that dauricine directly inhibited growth and proliferation of NB4 cells. Cells were incubated with 20 to 80 μmol/L dauricine after 24 h, the cell viability was decreased from （86.5 ± 11.7）% to （4.1 ± 0.5）%. In addition, cells were treated with 10 to 40μmol/L dauricine for 24 h. The flow cytometry analysis revealed that the cell apoptosis rate and ROS level was significantly increased and MMP was decreased, rl3~e AFM results indicated that normal cells were round and plump, the surface was smooth. Dauricine made the NB4 cell began to collapse, and the mean roughness of cells surface rised. [ Conclusion ] Dauricine directly inhibited proliferation and induced apoptosis of NBd cells.

关 键 词：蝙蝠葛碱 NB4细胞 原子力显微镜 超微结构 细胞凋亡 

分 类 号：R733.71[医药卫生—肿瘤] R285.5[医药卫生—临床医学]