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目的探讨杨梅素对人肝癌细胞系HepG2凋亡的影响及其机制。方法以不同浓度的杨梅素孵育HepG2细胞,在不同时间分别采用倒置显微镜观察细胞形态学变化;采用噻唑蓝(MTT)法检测细胞存活率;采用乳酸脱氢酶(LDH)活力定量测定试剂盒测定细胞上清液LDH活性;采用流式细胞术检测细胞凋亡率;采用蛋白质印迹法检测细胞凋亡途径关键蛋白的表达。结果杨梅素对HepG2细胞的凋亡诱导作用呈现剂量和时间依赖性。杨梅素作用于HepG2细胞,随杨梅素浓度增加,细胞凋亡率增加。与正常对照组相比,100μmol/L杨梅素作用于HepG2细胞24 h后,细胞存活率明显降低[(100.02±5.97)%vs.(71.60±3.75)%,P=0.006];早、晚期细胞凋亡率明显升高[(10.19±2.61)%vs.(2.52±2.05)%,P=0.003;(11.71±3.34)%vs.(3.69±1.13)%,P=0.004],差异均有统计学意义。随杨梅素浓度增加,线粒体途径的促凋亡因子BAX蛋白表达量增加,而抗凋亡因子BCL-2蛋白表达量降低。结论杨梅素能够诱导HepG2细胞凋亡,可能具有潜在的抗肝癌活性;线粒体途径在此过程中起重要作用。
Objective To investigate the effect of myricetin on the apoptosis of human hepatoma cell line HepG2 and its mechanism. Methods HepG2 cells were incubated with myricetin at different concentrations. Morphological changes were observed by inverted microscope at different time points. Cell viability was measured by MTT assay. Lactate dehydrogenase (LDH) activity assay kit Cell supernatant LDH activity; flow cytometry was used to detect the apoptosis rate; Western blotting was used to detect the expression of key proteins in apoptosis pathway. Results Myricetin induced apoptosis in HepG2 cells in a dose-and time-dependent manner. Myricetin acts on HepG2 cells, with the myricetin concentration increased, the rate of apoptosis increased. Compared with the normal control group, the survival rate of HepG2 cells treated with 100μmol / L myricetin for 24 h was significantly lower than that of the normal control group [(100.02 ± 5.97)% vs (71.60 ± 3.75)%, P = 0.006] (10.19 ± 2.61)% vs (2.52 ± 2.05)%, P = 0.003; (11.71 ± 3.34)% vs (3.69 ± 1.13)%, P = 0.004], and the differences were statistically significant Significance of learning. With the increase of myricetin concentration, the expression of pro-apoptotic factor BAX increased in mitochondria, while the expression of anti-apoptotic factor BCL-2 decreased. Conclusion Myricetin can induce apoptosis of HepG2 cells and may have potential anti-hepatoma activity. Mitochondrial pathway plays an important role in this process.