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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 TXF on HepG2 cells. Methods HepG2 cells were treated with 3 different doses (6, 12 and 24 μg / ml) of TXF. Normal HepG2 cells were used as a control. After cultured for 24 hours, the growth inhibition was determined by trypan blue exclusion and MTT assay. The morphology of HepG2 cells was observed, the ultrastructure of HepG2 cells was observed by transmission electron microscope, and the expression of Fas / FasL was detected by RT-PCR. Results The proliferation of HepG2 cells was inhibited by trypan blue exclusion assay (P <0.01). Different concentrations of TXF inhibited the growth of HepG2 cells in a dose - dependent manner (P <0.01). The results of inverted phase contrast microscope showed that the cytoplasm was vacuolized, the phenomenon of chromatin margination was observed by TEM, and vacuoles and other apoptosis appeared in the nucleolus. The relative expression of Fas / FasL mRNA was significantly increased by RT-PCR (P <0.01). Conclusion Different doses of TXF have inhibitory effects on HepG2 cells. The cytotoxic effect of 12 μg / ml TXF on HepG2 cells mainly induces apoptosis. TXF leads to apoptosis through the Fas / FasL pathway.