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目的:探讨荔枝核提取物对人神经母细胞瘤SH-SY5Y细胞的增殖抑制及至凋亡机制。方法:采用MTT法检测细胞活力,Hoechst 33258染色观察细胞形态改变;流式细胞术测定细胞凋亡率、细胞周期的变化及Fas的表达;活性检测试剂盒测定样品对细胞caspase活性的影响。结果:荔枝核提取物L2.3对SH-SY5Y细胞的增殖抑制活性呈时间、剂量双重依赖性,细胞内可见染色质固缩,出现凋亡小体。细胞凋亡率于处理48 h后显著增高,G0/G1及G2/M期细胞减少,S期细胞增多,Fas的表达明显增高,caspase-3、8、9的活性均时间依赖性地增高。结论:荔枝核提取物L2.3可通过诱导细胞凋亡、影响细胞周期分布而抑制SH-SY5Y细胞增殖,caspase-3、caspase-8、caspase-9及Fas蛋白可能参与了L2.3诱导SH-SY5Y细胞凋亡的通路。
Objective: To investigate the inhibitory effect of Litchi seed extract on the proliferation of human neuroblastoma SH-SY5Y cells and its mechanism of apoptosis. METHODS: Cell viability was measured by MTT assay, Hoechst 33258 staining was used to observe the changes in cell morphology, flow cytometry was used to determine apoptosis rate, cell cycle changes, and Fas expression. The activity assay kit was used to determine the effect of the sample on cell caspase activity. RESULTS: The inhibitory activity of Litchi seed extract L2.3 on SH-SY5Y cells was time- and dose-dependent. Chromatin condensation and apoptotic bodies appeared in the cells. The rate of cell apoptosis was significantly increased after 48 hours of treatment, the number of cells in G0/G1 and G2/M phases was decreased, the number of cells in S phase was increased, the expression of Fas was significantly increased, and the activities of caspase-3, 8 and 9 all increased in a time-dependent manner. CONCLUSION: Litch nuclear extract L2.3 can inhibit the proliferation of SH-SY5Y cells by inducing apoptosis and affecting cell cycle distribution. Caspase-3, caspase-8, caspase-9 and Fas proteins may be involved in L2.3-induced SH - SY5Y cell apoptosis pathway.