摘要
目的研究荷叶碱对人肝癌细胞株HepG2凋亡及其作用机制。方法采用CCK-8法观察荷叶碱对人肝癌细胞株HepG2生长增殖的影响;Hoechst33258染色观察HepG2细胞凋亡形态的变化;流式细胞术检测细胞凋亡率和周期的影响;Western Blot检测HepG2细胞凋亡相关蛋白的表达。结果荷叶碱对体外培养的人肝癌细胞株HepG2的生长具有抑制作用,且呈剂量、时间依赖性;Hoechst 33258染色可见实验组细胞固缩,凋亡小体形成,蓝色荧光加深;流式结果表示,荷叶碱可使HepG2细胞阻滞于G0/G1期,细胞凋亡率随药物浓度的增大亦呈增长趋势;Western Blot结果表明,荷叶碱以浓度梯度抑制NF-КB蛋白,Bcl-2蛋白的表达,促进Bax蛋白的表达。结论荷叶碱可抑制肝癌HepG2细胞的生长增殖,并在一定的范围内呈剂量、时间依赖性。荷叶碱可将人肝癌细胞株HepG2阻滞于G0/G1期,诱导其发生凋亡,凋亡机制可能与调节相关蛋白NF-КB、Bcl-2、Bax的表达有关。
Objective To investigate the effects of nuciferine on apoptosis of human hepatocellular carcinoma cell line HepG2 and its mechanism. Methods The proliferation of nuciferine on HepG2 cells was determined by the assay of cell counting kit-8（CCK-8）. The apoptotic morphology of HepG2 cells with Hoechst 33258 staining was observed under fluorescent microscope. Cell cycle distribution and apoptotic rate were evaluated by flow cytomety（FCM）.Western Blot analysis was used to detect the protein expression of apoptosis. Results Nuciferine could inhibit the proliferation of human hepatocellular carcinoma cell line HepG2 in a dose and time dependent manner. Hoechst33258 staining showed that the experimental group of cells shrank, apoptotic bodies formed, blue fluorescence deepened and flow cytometry（FCM） indicated that HepG2 cells were arrested in G0/G1 phase and the apoptotic rate increased with the increase of the concentration of nuciferine. Western Blot result showed that nuciferine inhibited the expression of NF-КB and Bcl-2 proteins while promoted the expression of Bax protein in a concentration-dependent manner. Conclusion Nuciferine could inhibit the growth and proliferation of HepG2 cells in a dose and time-dependent manner and induce the apoptosis of HepG2 cells. Also, HepG2 cells were arrested in G0/G1 phase. Furthermore,up-regulation of Bax protein as well as downregulation of NF-КB and Bcl-2 protein expression may be one of the important apoptotic inducing mechanisms.
出处
《中国药物警戒》
2017年第12期715-719,726,共6页
Chinese Journal of Pharmacovigilance
