毒黄素对HepG2细胞的毒性作用及其机制研究

The toxic effect and the mechanism of toxoflavin on HepG2 cells

目的研究不同剂量毒黄素(TXF)对HepG2细胞的毒性作用及机制。方法取3种剂量(6、12和24μg/ml)的TXF作用于HepG2细胞,以正常HepG2细胞作对照,培养24h后,台盼蓝拒染法和MTT法测定细胞生长抑制作用,倒置相差显微镜下观察HepG2细胞形态结构,透射电镜观察HepG2细胞超微结构变化,RT-PCR对Fas/FasL的表达作半定量检测。结果盼蓝拒染法检测,不同剂量的TXF对HepG2细胞均有抑制作用(P<0.01);MTT检测,不同剂量的TXF均可抑制HepG2细胞生长,并呈剂量依赖关系(P<0.01);倒置相差显微镜观察,可见细胞质空泡化;TEM观察细胞出现染色质边集现象,核仁内出现空泡等细胞凋亡现象;RT-PCR检测Fas/FasL的mRNA相对表达量,均呈现明显增高(P<0.01)。结论不同剂量TXF对HepG2细胞均有抑制作用,12μg/mlTXF对HepG2细胞毒性作用主要表现为诱导细胞凋亡。TXF可通过Fas/FasL途径导致细胞凋亡。

Objective To study the toxic effects and mechanism of different concentrations Toxoflavin（TXF） on HepG2 cells, and to provide the theory basis for the toxic effect of TXF on human being. Methods Using 1 % FBS as the con trol, TXF of 6, 12 and 24 μg/ml were co-incubated with HepG2 cells for 24 hours in vitro. Then the effect of growth inhibitors were surveyed by methods of Trypan blue dyeing and MTT. The morphosis of HepG2 was observed by inverted phase contrast microscope. The ultrastructural organization of HepG2 cells were observed by TEM. Lastly, the expression of Fas/FasL mRNA was detected by RT PCR. Results The growth of HepG2 cells was obviously inhibited. TXF inhibited the proliferation of HepG2 cells in a dose-dependent-manner （P〈0.01）. Cytoplasm vacuolization could be observed by inverted phase contrast microscope. By TEM we could observe the vacuole emerged in nucleolus and chromatin rnargination. On the whole, the morphosis of HepG2 cells apoptosis was observed and TXF could induced the expression of Fas/Fast. mRNA（P〈0.01）. Conclusion TXF can inhibit the growth of HepG2 cells and TXF can also induce cell apoptosis by Fas/FasL. way.