荔枝核提取物对HepG-2细胞生长抑制及凋亡诱导机制的探讨

Antiproliferative and induced apoptosis effects of Litchi seed extract in HepG-2 cells

目的:探讨荔枝核提取物成分L2.3对人肝癌细胞HepG-2的生长抑制作用及其分子机制。方法:MTT法测定样品对HepG-2细胞的增殖抑制,荧光染色观察细胞凋亡;比色法检测样品对HepG-2中caspase-3、caspase-8和caspase-9活性的影响,流式细胞仪检测并分析Fas蛋白表达的变化。结果:L2.3对HepG-2细胞表现出时间及剂量依赖性增殖抑制活性(P<0.05),其半数抑制质量浓度为43.0μg/mL;处理组细胞中出现致密浓染亮蓝色凋亡小体。较高质量浓度的L2.3处理细胞后,caspase-3、8、9的活性均呈时间依赖性增高(P<0.01),且Fas蛋白的表达也明显上调(P<0.05)。结论:荔枝核提取物L2.3对HepG-2细胞有体外增殖抑制活性,并介导caspase-3、caspase-8、caspase-9活性改变及Fas蛋白表达的上调,以此诱导细胞凋亡的发生。

Aim：To detect the effect of Litchi seed extract L2.3 on inhibiting the proliferation of human hepatocellular carcinoma cell line HepG-2 cells and its mechanism on inducing cell apoptosis. Methods： MTT assay was used to test the antiproliferative effect of L2.3,and the cell apoptosis was observed after stained with Hoechst 33258.The activities of caspase-3、caspase-8、caspase-9 were determined by caspase colorimetric assay kit.Flow cytometry（FCM） analysis was carried out to examine the expression of Fas protein in HepG-2 cells.Results： Litchi seed extract L2.3 showed dose-and time-dependent antiproliferative effect on HepG-2 cells（P0.05）.The 50% inhibitory concentration（IC50） of L2.3 was 43.0 μg/mL.In Hoechst 33258 stained cell,there were karyopyknosis and some apoptotic body with thick bright blue fluorescence were observed.The activities of caspase-3、caspase-8、caspase-9 were increased with time-dependent manner（P0.01）.After treated with high concentration of L2.3 for 48 h,Fas protein expressed in HepG-2 cells showed a significant increase.Conclusion：Litchi seed extract L2.3 can inhibit the proliferation of HepG-2 cells in vitro and induce cell apoptosis.Activation of caspase-3、caspase-8、caspase-9 change and Fas expression increase may be involved in the pathway of apoptosis in HepG-2 cells treated with L2.3.