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目的考察灰树花提取物抗肝癌作用及相关分子机制。方法在PLC/PRF/5和HepG2细胞中,采用灰树花提取物处理给药法,噻唑蓝法测定细胞存活率,化学法测定乳酸脱氢酶释放及Capase3活力,JC-1染色测定线粒体跨膜电位变化,Western Blot测定Bcl-2和Bax蛋白表达以及Akt/GSK3β磷酸化情况。在PLC/PRF/5异位荷瘤裸鼠模型中,灰树花的抗肝癌效果得到进一步验证。结果灰树花提取物可以有效降低PLC/PRF/5和HepG2细胞存活率,提高细胞乳酸脱氢酶释放,增加Caspase3活力,减弱线粒体跨膜电位,增加Bax表达,降低Bcl-2蛋白含量,降低Akt/GSK3β磷酸化水平。在不改变体重的情况下,灰树花可有效抑制在PLC/PRF/5异位荷瘤裸鼠模型中肿瘤的生长。结论灰树花可通过线粒体依赖途径及Akt/GSK3β通路有效起到抗肝癌活性。
Objective To investigate the anti-hepatocarcinoma effect and the related molecular mechanism of Grifola frondosa extract. Methods The cell viability was measured by MTT in PLC / PRF / 5 and HepG2 cells, the cell viability was assayed by thiazolyl blue method, the lactate dehydrogenase release and Capase3 activity were measured by chemical method, Membrane potential changes, Western Blot determination of Bcl-2 and Bax protein expression and phosphorylation of Akt / GSK3β. In PLC / PRF / 5 ectopic tumor-bearing nude mice models, Grifola frondosa anti-liver cancer effect was further verified. Results Grifola frondosa extract can effectively reduce the survival rate of PLC / PRF / 5 and HepG2 cells, increase the release of cellular lactate dehydrogenase, increase the activity of Caspase3, decrease the transmembrane potential of mitochondria, increase the expression of Bax, decrease the content of Bcl-2 and decrease Akt / GSK3β phosphorylation level. Without changing body weight, Grifola frondosa can effectively inhibit tumor growth in a nude mouse model of PLC / PRF / 5 ectopic tumor. Conclusion Grifola frondosa can effectively inhibit liver cancer activity through mitochondria-dependent pathway and Akt / GSK3β pathway.