麦麸结合态多酚对肝癌HepG-2细胞增殖及凋亡效应研究

目的 本文主要研究麦麸结合态多酚对肝癌HepG-2细胞的抑制效应。方法 小麦麸皮粉碎后,采用丙酮-碱消化法,获得麦麸结合态多酚物质,命名为：WBBP,用福林酚比色法测定该多酚的含量及提取得率；通过MTT法检测WBBP处理对肝癌HepG-2细胞增殖的抑制效应,倒置显微镜观察WBBP处理对HepG-2肝癌细胞形态学的影响；采用流式细胞仪检测WBBP处理对HepG-2肝癌细胞周期、凋亡情况以及线粒体膜电位的影响；利用caspases试剂盒检测WBBP作用下肝癌HepG-2细胞中Caspase-3、Caspase-8、Caspase-9的酶活性变化。结果显示,不同浓度的WBBP通过诱导HepG-2肝癌细胞周期在S期阻滞,显著抑制了细胞增殖,且具有浓度依赖性；同时WBBP通过caspase依赖的线粒体凋亡路径诱导HepG-2肝癌细胞凋亡率以浓度依赖的方式增加。结论 麦麸结合态多酚WBBP能够通过显著抑制HepG-2肝癌细胞的增殖,并诱导细胞凋亡来发挥抗肝癌效应。

Effects of bound polyphenols from wheat bran on cell proliferation and apoptosis of HepG-2 cells

Objective Investigating the inhibitory effect of bound polyphenols from wheat bran on hepatocellular carcinoma (HCC) cells. Methods The bound polyphenols were obtained by acetone-alkali digestion method and then extracted ethylacetate after wheat bran was crushed, which were designated as WBBP; the content and extraction rate of WBBP were measured by Folin-Ciocalteu assay; the inhibitory effect of WBBP on HepG-2 cells was detected by MTT method. Morphological changes of HepG-2 cells treated with WBBP were observed by optical microscope. Flow cytometry was used to detect the effects of WBBP on the cell cycle, apoptosis and mitochondrial membrane potential of HepG-2 cells. Effects of WBBP on the enzyme activities of caspase-3, Caspase-8 and Caspase-9 were detected by the corresponding kits. Results Compared with the control group, the different concentrations of WBBP significantly inhibited cell proliferation of HepG-2 through inducing cell cycle in the S phase arrest. Meanwhile, WBBP could dose-dependently induce cell apoptosis of HepG-2 via caspase-dependent mitochondrial apoptosis pathway. Conclusion The anti-HCC activity of WBBP attributed to the inhibition of cell proliferation and inducion of cell apoptosis of HepG-2.