目的旨在研究α-维尼非林(α-viniferin)对人慢性髓系白血病细胞K562的作用与相关机制。方法 MTT法评价α-viniferin对K562细胞的细胞毒活性。采用细胞形态学和生物化学方法检测细胞凋亡。通过化学荧光法对细胞内线粒体膜电位、caspase-9、caspase-3活性分析。通过半定量RT-PCR表达分析来确定Bcl-2家族相关基因在α-viniferin诱导K562细胞凋亡中的作用。结果α-viniferin能抑制K562细胞增殖,呈剂量和时间依赖性, IC50为13.61 mg · L-1。α-viniferin引起K562细胞出现死亡并伴随有染色质聚集、核破碎、凋亡小体等典型的凋亡形态学特征;此外还伴随有线粒体膜电位显著降低,caspase-9、caspase-3活性升高等现象;α-viniferin(2~32 mg·L-1)引起K562细胞caspase-3 mRNA表达持续升高,Bax、Bad、Bim、Bid促凋亡基因mRNA表达增加,而Bcl-2、Bcl-xL抗凋亡基因mRNA表达持续下降。结论α-viniferin通过线粒体途径诱导K562细胞凋亡。
Abatract:Aim To investigate the effect and disclose its potential mechanisms of α-viniferin-induced prolif-eration and apoptosis in chronic myeloid leukemia K562 cells. Methods K562 cells were exposed to α-viniferin. The cytotoxicity was analyzed by MTT assay. The apoptosis in K562 cells was proved by both cellu-lar morphological and biochemical methods. The chem-ical fuorescence assays were employed to evaluate the mitochondrial membrane potential, the relative changes of the caspase-9 and caspase-3 in the α-viniferin-trea-ted K562 cells. The Bcl-2 family related gene expres-sion was analyzed by semi-quantitative RT-PCR. Re-sults After α-viniferin treatment, the proliferation of K562 cells was inhibited in a dose-dependent and time-dependent manner. The cells were treated with 32 mg · L-I α-viniferin and showed significant apoptotic morphological changes with chromatin aggregation, nu- clear fragmentation and apoptotic bodies coming into being. The JC-1 fluorescence stain showed significant decreases in green/red ratio in α-viniferin treated groups. Further, caspase-9 and caspase-3 activities were increased. Bax and Bcl-xL relative mRNA level was down-regulated, and there was up-regulation of Bax and so on. Conclusion α-viniferin inhibits the proliferation and induces mitochondrial pathway. apoptosis in K562 cells via