目的:探讨小叶莲提取物、有效成分及成分组合对人乳腺癌细胞增殖的影响及机制.方法:采用酸性磷酸酶法测定小叶莲4种提取物[乙醇提取物(Xc)、乙醇提取物石油醚萃取物(Xp)、乙醇提取物乙酸乙酯萃取物(Xe)、乙醇提取物正丁醇萃取物(Xz)]、5种有效成分[鬼臼毒素(S1)、去氧鬼臼毒素(S2)、4-去甲去氧鬼臼毒素(S3)、8-异戊烯基山柰酚(S4)、8,2′-二异戊烯基槲皮素-3-甲醚(S5)]以及3种有效成分组合{组合1[S1-S2-S3-S4-S5(2:4:1:4:32),Z1]、组合2[S2-S4(1:1),Z2]、组合3[S3-S4(1:4),Z3]}对人乳腺癌MDA-MB-231、MCF-7细胞增殖的影响;流式细胞仪检测上述样品对MDA-MB-231、MCF-7(或T47D)细胞周期和细胞线粒体膜电位的影响.结果:小叶莲有效成分组合Z1对MDA-MB-231、MCF-7细胞具有显著抑制作用,其半数抑制浓度(IC50)分别为(0.27±0.2)、(0.11±0.1)μg/mL;提取物Xc、Xp、Xe,有效成分S2、S4以及成分组合Z2、Z3对MDA-MB-231、MCF-7(或T47D)细胞的增殖均有较强抑制作用,IC50均小于15μg/mL;各提取物和有效成分均可阻滞MDA-MB-231、MCF-7细胞周期于G2/M期;各有效成分组合均可阻滞MDA-MB-231、T47D细胞周期于G0/G1期,并能降低MDA-MB-231、T47D细胞线粒体膜电位.结论:小叶莲有效成分及成分组合可通过影响细胞周期和线粒体膜电位,从而抑制乳腺癌细胞的增殖.
OBJECTIVE:To explore the effects and mechanism of extracts,active constituents and constituent combination of Sinopodophylli Fructus on cell proliferation of human breast cancer. METHODS:Acid phosphatase method was conducted to deter-mine the effects of 4 extracts [ethanol extract (Xc),petroleum ether extract from ethanol extract (Xp),ethyl acetate extract from ethanol extract (Xe),n-butanol extract from ethanol extract (Xz)],5 active constituents [podophyllotoxin (S1),deoxypodophyllo-toxin (S2),4-desmethyl deoxypodophyllotoxin (S3),8-isopentenyl kaempferol (S4),8,2′-diisoprenyl quercetin-3-methyl ether (S5)] and 3 active constituent combination [combination 1,S1-S2-S3-S4-S5 (2:4:1:4:32),Z1;combination 2,S2-S4 (1:1),Z2;combination 3,S3-S4(1:4),Z3] on the MDA-MB-231,MCF-7 cell proliferation;flow cytometry was adopted to detect the effects of above-mentioned samples on MDA-MB-231,MCF-7(T47D)cell cycle and mitochondrial membrane potential. RESULTS:The active constituent combination Z1 showed significant inhibitory effects on MDA-MB-231,MCF-7 cells,the half inhibitory concen-trations(IC50)were(0.27±0.2),(0.11±0.1)μg/mL;extracts Xc,Xp,Xe,active constituents S2,S4 and active constituent combi-nation Z2,Z3 showed relatively strong inhibitory effects on MDA-MB-231,MCF-7 (T47D) cell proliferation (IC50<15 μg/mL). Both extracts and active constituents can block MDA-MB-231,MCF-7 cell cycle in G2/M phase;all active constituents can block MDA-MB-231,T47D cell cycle in G0/G1 phase,and can reduce MDA-MB-231,T47D cell mitochondrial membrane potential. CONCLUSIONS:The active constituents and constituent combination of Sinopodophylli Fructus can inhibit cell proliferation of breast cancer by affecting cell cycle and mitochondrial mem-brane potential.