葡萄籽原花青素诱导人肺癌A549细胞凋亡的研究

Study on the Effect of Grape Seed Proanthocyanidins on the Apoptosis of Human Lung Carcinoma A549 Cells

[目的]研究葡萄籽提取物原花青素对人肺癌细胞株A549凋亡的影响.[方法]以不同浓度的原花青素与 A549细胞体外共同培养,采用噻唑蓝（MTT）法测定细胞增殖抑制率;Hoechst33258染色观察凋亡细胞形态学变化;TUNEL法检测细胞凋亡率.[结果]原花青素可体外抑制 A549细胞生长,在2.5~250 μmol/L的浓度范围,细胞增殖抑制率差异极为显著（ P 〈0.01）.作用24 h的IC50值达7.40 μmol/L,且呈浓度和时间依赖性;荧光染色可见不同浓度原花青素作用于细胞后,凋亡细胞增加,并可见染色质凝集、细胞核碎裂等细胞凋亡特征性变化.TUNEL法计量结果显示,随着PA作用浓度的增加,凋亡率增高,呈剂量依赖性.实验组细胞凋亡率均高于对照组（ P 〈0.05）.[结论]原花青素在体外能通过抑制人肺癌细胞株A549细胞增殖,促进细胞凋亡.

[Objective]To explore the effect of grape seed extract proanthocyanidins（PA） on the apoptosis of human lung carcinoma A549 cells. [Methods] The A549 cells were cultivated with different concentration of PA. The suppression rate of cell proliferation was measured by MTT assay. Cell apoptosis was observed by fluorescence microscope techniques with Hoechst 33258 assay. TUNEL method was used to detect the cell ap- optosis rate, [Results]PA could inhibit the proliferation of A548 cells. While the concentration of PA was 2. 5 ~250 μmol/L, there was most significant difference in the suppression rate of cell proliferation （ P 〈0.01）. The ICS0 for 24h was up to 7.40 μmol/L in a dose- and time-dependent manner. Fluorescence staining showed that the apoptotic cells increased and the apoptotic characteristic changes such as chromatic agglutination and nuclear fragmentation were found after the cells were cultivated with different concentrations of PA on cells. TUNEL assay showed that the apoptosis rate increased with the concentration of PA in a dose-dependent man- ner. The apoptosis rate of experimental group was higher than that of control group（ P 〈0.05）. [Conclusion] PA can promote the cell apoptosis by inhibiting the proliferation of human lung carcinoma A549 cells in vitro.