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目的:探讨八角中莽草酸对人肝癌细胞HepG-2细胞增殖及其核因子-κB(NF-κB)p65蛋白表达的影响。方法:用不同浓度的莽草酸处理HepG-2细胞,MTT法测定莽草酸对HepG-2细胞的生长抑制率;倒置显微镜下观察莽草酸作用HepG-2细胞后的形态学变化;采用hoechest33342荧光染色观察莽草酸作用于人肝癌HepG-2细胞48 h后细胞凋亡形态变化。Western blot试验观察莽草酸对人肝癌HepG-2细胞NF-κB(p65)蛋白表达的影响。结果:MTT实验结果表明,莽草酸对人肝癌HepG-2细胞的生长抑制作用呈现时间依赖性和剂量依赖性;倒置显微镜下观察细胞形态可以发现,经莽草酸处理48 h后的各组细胞中,可观察到随着药物浓度的增加,细胞凋亡形态变化越来越明显,细胞数量逐渐变少。荧光染色观察随着药物浓度增加,凋亡细胞增多且明显,当莽草酸质量浓度达1 g·L-1时,出现较多的凋亡小体。NF-κB(p65)蛋白的表达随莽草酸浓度升高而显著下降。结论:莽草酸对人肝癌细胞HepG-2有较显著的生长抑制作用,莽草酸对人肝癌HepG-2细胞表达NF-κB(p65)明显减弱,其诱导人肝癌HepG-2细胞凋亡作用机制可能下调NF-κB表达水平有关。
AIM: To investigate the effects of shikimic acid on the proliferation and expression of nuclear factor-κB (p65) protein in HepG-2 cells. Methods: HepG-2 cells were treated with different concentrations of shikimic acid. The growth inhibition rate of shikimic acid on HepG-2 cells was determined by MTT assay. The morphology of HepG-2 cells was observed under inverted microscope. The hoechest33342 fluorescence staining The changes of apoptosis of HepG-2 cells treated with shikimic acid for 48 h were observed. The effect of shikimic acid on the protein expression of NF-κB (p65) in HepG-2 cells was observed by Western blot. Results: The results of MTT assay showed that the inhibitory effect of shikimic acid on HepG-2 cells was time-dependent and dose-dependent. Under inverted microscope, the morphological changes of cells were observed. After treated with shikimic acid for 48 h, It can be observed that with the increase of drug concentration, the morphological changes of apoptotic cells become more and more obvious, and the number of cells gradually decreases. Fluorescence staining showed that apoptotic cells increased and obviously with the increase of drug concentration. When shikimic acid concentration reached 1 g · L-1, more apoptotic bodies appeared. The expression of NF-κB (p65) protein decreased significantly with increasing shikimic acid concentration. Conclusion: Shikimic acid has a significant inhibitory effect on HepG-2 cells. Shikimic acid attenuates the expression of NF-κB (p65) in HepG-2 cells and induces the apoptosis of HepG-2 cells. May be related to down-regulation of NF-κB expression.