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目的:观察来源于海芒果种子的皂苷类化合物nerifolin对人肝癌细胞HepG2增殖及凋亡的影响,初步探讨其作用机制。方法:MTT法检测不同质量浓度nerifolin对HepG2细胞增殖的影响,流式细胞术检测nerifolin对HepG2细胞周期和凋亡的影响。Caspase试剂盒检测nerifolin对HepG2细胞caspase-3酶活化的影响。结果:Nerifolin可时间和剂量依赖性地抑制HepG2细胞增殖,作用24、48、72h的IC50分别为(2.34±0.08)、(0.13±0.01)、(0.06±0.01)μg/ml。随着nerifolin(0.1μg/ml)作用时间的延长,HepG2细胞S期百分比逐渐增多,而G0/G1期百分比逐渐减少(P<0.01),nerifolin阻滞HepG2细胞于S期。Nerifolin作用后,HepG2细胞早期凋亡率上升至22.65%,caspase-3活性比对照组明显升高(P<0.01)。结论:Neri-folin可通过S期阻滞抑制HepG2细胞的增殖,通过caspase-3依赖途径诱导HepG2细胞的凋亡。
OBJECTIVE: To observe the effect of nerifolin derived from sea mango seeds on the proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2, and to explore its mechanism. Methods: The effect of different concentrations of nerifolin on the proliferation of HepG2 cells was detected by MTT assay. The effect of nerifolin on the cell cycle and apoptosis of HepG2 cells was detected by flow cytometry. Caspase kit assay nerifolin on HepG2 cells caspase-3 enzyme activation. Results: Nerifolin could inhibit the proliferation of HepG2 cells in a time and dose-dependent manner. The IC50 values at 24, 48 and 72 hours were (2.34 ± 0.08), (0.13 ± 0.01) and (0.06 ± 0.01) μg / ml, respectively. The percentage of S phase of HepG2 cells gradually increased, while the percentage of G0 / G1 phase decreased gradually (P <0.01) with the increase of nerifolin (0.1μg / ml), and nerifolin blocked HepG2 cells in S phase. After Nerifolin treatment, the early apoptosis rate of HepG2 cells increased to 22.65%, and the activity of caspase-3 was significantly higher than that of the control group (P <0.01). Conclusion: Neri-folin can inhibit the proliferation of HepG2 cells through S-phase arrest and induce the apoptosis of HepG2 cells through a caspase-3-dependent pathway.