山楂原花青素对人肝癌SMMC-7721细胞增殖和凋亡的影响

Effect of Hawthorn Proanthocyanidins on Proliferation and Apoptosis of Human Hepatocellular Carcinoma SMMC-7721 Cells

目的:探讨山楂原花青素提取物、表儿茶素和原花青素B2对人肝癌SMMC-7721细胞增殖及凋亡的影响。方法:体外培养SMMC-7721细胞,噻唑蓝法检测不同质量浓度山楂原花青素对细胞增殖的影响;4’,6-二脒基-2-苯基吲哚染色法观察细胞凋亡,Western blot法检测细胞凋亡相关蛋白的表达;检测每组细胞中过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、Na^+K^+-ATP酶活力和丙二醛(malondialdehyde,MDA)含量。结果:山楂原花青素提取物、表儿茶素和原花青素B_2呈时间和质量浓度依赖性地显著抑制SMMC-7721细胞的增殖(P<0.05),促进细胞凋亡;与对照组相比,SMMC-7721细胞中SOD、CAT、GSH-Px、Na^+K^+-ATP酶活力降低,MDA含量升高;细胞凋亡相关蛋白半胱天冬酶-8、半胱天冬酶-3、Bcl-2相关X蛋白表达量和多聚二磷酸腺苷核糖聚合酶片段化增加,Bcl-2表达量降低。结论:山楂原花青素可通过影响抗氧化酶活力和细胞凋亡相关蛋白来抑制SMMC-7721细胞增殖,促进细胞凋亡。

Objective:To investigate the effect of hawthorn proanthocyanidins extract,epicatechin and proanthocyanidin B2 on the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cells.Methods:SMMC-7721 cells were cultured in vitro.The effect of different concentrations of hawthorn proanthocyanidins on cell proliferation and apoptosis was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay and 4’,6-diamidino-2-phenylindole(DAPI)staining,respectively.The expression of apoptosis-related proteins was detected by Western blotting.The activities of catalase(CAT),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and Na+K+-ATPase and malondialdehyde(MDA)level in cells from each group were determined.Results:Hawthorn proanthocyanidins extract,epicatechin and proanthocyanidin B2 all significantly inhibited the proliferation of SMMC-7721 cells in a time-dependent and concentrationdependent manner(P<0.05)and induced cell apoptosis.Compared with the control group,hawthorn proanthocyanidins extract,epicatechin and proanthocyanidin B2 decreased the activity of SOD,CAT,GSH-Px and Na+K+-ATPase and increased the level of MDA in SMMC-7721 cells.The expression of apoptosis-related proteins casepase-8,caspase-3,Bcl-2-associated X protein and the fragment of poly(ADP-ribose)polymerase were increased,while the expression of Bcl-2 was decreased.Conclusion:Hawthorn proanthocyanidins can inhibit the proliferation of SMMC-7721 cells and induce apoptosis through regulating antioxidant activity and apoptosis-related proteins.