山奈酚对HepG2细胞增殖的影响及诱导其凋亡的机制研究

目的研究山奈酚对人肝癌细胞(HepG2细胞)增殖与凋亡的影响及其机制。方法将HepG2细胞分为空白组和实验组,实验组以CCK-8法检测10个不同浓度的(10,20,30,40,50,60,70,80,90,100μmol·L^-1)山奈酚处理HepG2细胞24 h后的存活率。最终以3个浓度(20,40,50μmol·L^-1)山奈酚作为低、中、高3个浓度实验组。以免疫印迹法检测核苷酸结合寡聚化域样受体蛋白3(NLRP3)和死骨片-1(P62)蛋白表达水平(灰度值)。结果山奈酚处理HepG2细胞24 h后,空白组与低、中、高3个浓度实验组HepG2细胞的存活率分别为(100.00±0.00)%,(87.92±3.13)%,(77.92±4.40)%和(70.53±4.19)%;上述这4组的NLRP3表达水平分别为0.27±0.05,0.50±0.03,0.71±0.08和0.93±0.10;上述这4组的P62表达水平分别为0.54±0.06,0.76±0.05, 0.87±0.04和1.09±0.10。上述指标:3个浓度实验组与空白组比较,差异均有统计学意义(P<0.05,P<0.01,P<0.001)。结论山奈酚可有效抑制HepG2细胞增殖、抑制其自噬并诱导其凋亡,其机制可能为有效促进P62的沉积,同时能诱导凋亡经典途径中NLRP3的合成,从而抑制HepG2细胞自噬并促进其凋亡。

Effect of kaempferol on the proliferation of HepG2 cells and the mechanism of inducing apoptosis of HepG2 cells

Objective To research the effects of kaempferol on the proliferation and apoptosis of human hepatocellular carcinoma cells(HepG2 cells) and to explore its mechanism. Methods HepG2 cells were divi-ded into two groups:blank group and experimental group. The survival rate in the experimental group was treated with ten different concentrations(10, 20, 30, 40, 50, 60, 70, 80, 90, 100 μmol·L^-1) of kaempferol for 24 hours and detected by CCK-8 method. Finally, three concentrations(20,40,50 μmol ·L^-1)of kaempferol were used as the experimental-L,experimental-M,experimental-H groups. The expression level(grey ralue) of the nucleotide binding oligomerization domain-like receptor protein 3(NLRP3) and Dead bone slices-1(P62) in cultured HepG2 cells were detected by Western blotting. Results HepG2 cells treated with kaempferol for 24 hour,the survival rates of blank group and experimental-L,experimental-M,experimental-H groups were(100.00±0.00)%,(87.92±3.13)%,(77.92±4.40)% and( 70. 53 ± 4. 19) %,respectively;the expression levels of NLRP3 in the above four groups were 0. 27 ± 0. 05,0. 50 ± 0. 03,0. 71 ± 0. 08 and 0. 93 ± 0. 10,respectively;the expression levels of P62 in the above four groups were0. 54 ± 0. 06,0. 76 ± 0. 05,0. 87 ± 0. 04 and 1. 09 ± 0. 10,respectively. Comparison between the three experimental groups and blank group,the difference of the factors were statistically significant( P < 0. 05,P < 0. 01,P < 0. 001).Conclusion Kaempferol can effectively inhibit the proliferation of HepG2 cells,inhibit autophagy and induce the pyroptosis of HepG2 cells. The mechanism may be that kaempferol can effectively promote the deposition of P62 and induce the NLRP3 in the classical pathway of death of HepG2 cells,thus inhibiting the autophagy of HepG2 cells and promoting the pyroptosis of HepG2 cells.