线粒体参与银杏酸诱导HepG2细胞凋亡的研究  

Mitochondrial is involved in the apoptosis of HepG2 Cells induced by Ginkgolic acids

作　　者：叶思思[1] 高静[1] 龚爱华[2] 张严[2] 王穆彬[2] 杨小明[3] 

机构地区：[1]江苏大学药学院,镇江212013 [2]江苏大学基础医学与医学技术学院,镇江212013 [3]江苏大学食品与生物工程学院,镇江212013

出　　处：《南京大学学报（自然科学版）》2012年第3期360-366,共7页Journal of Nanjing University（Natural Science）

摘　　要：探讨银杏酸(Ginkgolic acids,GAs)诱导人肝癌细胞HepG2凋亡的效应,初步探索其与线粒体相互作用的机制.以不同浓度的GAs作用于HepG2,MTT检测与相差显微镜观察相结合,分析GAs对细胞增殖的影响;Hochest染色观察细胞核变化;AnnexinⅤ/PI双标后流式细胞仪检测凋亡情况;Western blot检测凋亡相关蛋白(Caspase-3、Bax、Bcl-2)的表达;JC-1染色检测线粒体膜电位;DCFH-DA染色检测活性氧(ROS)生成.结果发现,GAs以时间和剂量依赖性方式抑制细胞的增殖;GAs处理诱导HepG2细胞发生凋亡:细胞核皱缩及片段化,Caspase-3活化,Bax/Bcl-2比值增加,细胞凋亡率显著上升.此外,GAs可直接与线粒体发生作用,诱导HepG2细胞线粒体外膜膜电位下降,PTP孔道开放,ROS含量升高.研究结果表明GAs能够诱导肝癌细胞的凋亡,推测其主要通过影响线粒体功能而导致细胞的凋亡.The anticancer effect of Ginkgolic acids （GAs） was investigated in human liver carcinoma cell line HepG2, and the possible mechanism was analyzed. Cancer ceils proliferation were detected by MTT assay and the 50% inhibitory concentration（IC50） were calculated; nucleus morphological changes were observed by stained with hoeehest 33342; Cell apotosis rate was detected by flow eytometry （FCM） following Annexin V/P1 staining; expression of apoptosis-related protein Caspase-3, Bax and Bcl-2 were analysed by western blot; mitochondria swelling was used to find out its direct interaction with mitochondria; mitochondrial membrane potential was observed by JC-1 staining; the formation of ROS were detected by DCFH-DA staining. It was found that GAs inhibited proliferation of HepG2 cells in a dose-and time-dependent way. GAs induced apotosis indicated by chromatin condensation, Caspase-3, Bax activation. Meanwhile, （;-As was found to inhibit Ca^2＋-induced mitochondria swelling, decrease mitochondrial membrane potential and increase intracellular ROS level. In summary, GAs could inhibit the proliferation of cancer cells effectively. Its mechanism might be related to induction of apoptosis and mitochondrial regulation was involved in cell death process.

关 键 词：银杏酸 抗肿瘤 凋亡 线粒体 

分 类 号：Q513[生物学—生物化学]