摘要
目的:研究蓝莓花青素(Blueberry Anthocyanin,BA)诱导人胃腺癌BGC-823细胞凋亡过程中线粒体超微结构、线粒体膜电位及凋亡调节蛋白的改变。方法:采用倒置相差显微镜观察细胞形态;末端脱氧核苷酸转移酶介导的dU TP缺口末端标记(TUNEL)法检测细胞凋亡;透射电镜法观察线粒体超微结构改变;原位末端标记/碘化吡啶(TUNEL/PI)双染流式细胞术检测细胞凋亡率;罗丹明123(Rhodamine 123,Rho123)荧光探针标记流式细胞仪检测和分析线粒体跨膜电位(MTP,△Ψm);蛋白免疫印迹法检测细胞凋亡调节相关蛋白Bax和Bcl-2的表达水平。结果:蓝莓花青素呈剂量和时间依赖性的抑制人胃腺癌BGC-823细胞生长;100μg/ml蓝莓花青素作用4h后线粒体肿胀空泡化、内部结构消失,8h后细胞核染色质成块状边集,细胞凋亡;不同浓度蓝莓花青素作用24h后,流式细胞仪检测BGC-823细胞凋亡率呈浓度依赖性增加。不同浓度蓝莓花青素作用BGC-823细胞24h后,代表线粒体膜电位的Rho123荧光强度降低;经100μg/ml蓝莓花青素刺激BGC-823细胞4、12、24小时后促凋亡蛋白Bax表达升高,抑凋亡蛋白Bcl-2表达降低。结论:蓝莓花青素对人胃腺癌BGC-823细胞具有生长抑制和诱导凋亡作用,诱导凋亡可能通过线粒体凋亡途径起作用。
Objective： To study the ultrastructural and fuctional change of mitochondrial,and the change of apoptosis modulating proteins in the process of apoptosis of human gastric adenocarcinoma BGC-823 cell induced by Blueberry Anthocyanin. Methods： Phase contrast microscope was used to observe the cells morphology changed; TUNEL staining flow Cytometry method was used to observe the apoptosis of the BGC-823cells; The ultrastructure changes of BGC-823 cells were observed with transmission electron microscope; The mitochondria transmembrane potential（ MTP,△Ψm） change was calculated by Rhodamine 123（ Rho123） fluorescence probe was measured by flowcytometer; The expression changes of Bax and Bcl-2 in BGC-823 cells induced by Blueberry Anthocyanin were detected by Western Blot. Results： Blueberry Anthocyanin significantly inhibited the proliferation of BGC-823 cells in dose and time dependent manner. In the early-stage（ 4h） after adding Blueberry Anthocyanin,the mitochondria swelled with outer membrane broken; Then after 8 hours,the cell nucleus showed typical apoptotic changes. After 24 hours of adding Blueberry Anthocyanin,the cell apoptosis rate was enhanced in a concentration-dependent manner. In contrast,the fluorescent intensity decrease of the mitochondrial transmenbrane potential in a concentration-dependent manner. The ratio of Bax /Bcl-2 in BGC-823 cell were increased after adding 100μg / ml of Blueberry Anthocyanin for 24 hours. Conclusion： Blueberry Anthocyanin could inhibit the proliferation of BGC-823 cells and induce apoptosis of BGC-823 cells in time- and dose-dependent manner in vitro. Blueberry Anthocyanin induce apoptosis of BGC-823 cells may through mitochondrial-dependent pathway.
出处
《中药药理与临床》
CAS
CSCD
北大核心
2015年第4期56-60,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
河南大学校内基金[2013YBZR021]
关键词
蓝莓花青素
人胃腺癌
线粒体
膜电位
细胞凋亡
blueberry anthocyanin（蓝莓花青素）
human gastric adenocarcinoma
mitochondrial
membrane potential
apoptosis
