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目的探讨大豆异黄酮(SI)代谢物雌马酚对血管内皮细胞凋亡和增殖的影响。方法体外培养血管内皮细胞(ECV-304),采用不同浓度雌马酚(0、6.25、12.50、25.00、50.00和100.00μmol/L)处理24 h,收集细胞,采用流式细胞术分析细胞内活性氧(ROS)、细胞凋亡和细胞周期;采用免疫组织化学技术分析凋亡相关蛋白Bax、Bcl-2和caspase-3表达;采用噻唑蓝(MTT)法分析雌马酚对ECV-304细胞增殖影响。结果与对照组比较,6.25μmol/L雌马酚组ECV-304细胞内ROS含量下降,100.00μmol/L雌马酚组ECV-304细胞内ROS含量升高;与对照组比较,25.00、50.00、100.00μmol/L雌马酚组ECV-304细胞早、晚期凋亡率明显升高,并可导致细胞G1期阻滞(P<0.05);MTT实验结果显示,0、6.25、12.50、25.00、50.00、100.00μmol/L雌马酚组ECV-304细胞增殖抑制率分别为-(8.23±2.21)%、-(3.29±1.07)%、(6.75±1.73)%、(25.46±4.82)%和(36.93±4.66)%,低剂量雌马酚对细胞生长具有促进作用,高剂量雌马酚对细胞生长具有抑制作用;与对照组比较,25.00μmol/L雌马酚组作用24 h后,细胞内Bax和caspase-3表达增加,而Bcl-2表达减少。结论低浓度雌马酚可降低细胞内ROS含量,促进ECV-304细胞增殖;高浓度雌马酚可导致细胞凋亡、细胞周期阻滞,抑制细胞增殖。
Objective To investigate the effect of soybean isoflavone (SI) metabolite equol on apoptosis and proliferation of vascular endothelial cells. Methods Vascular endothelial cells (ECV-304) were cultured in vitro and treated with different concentrations of equol (0, 6.25,12.50,25.00,50.00 and 100.00 μmol / L) for 24 h. Cells were collected and analyzed by flow cytometry (ROS), apoptosis and cell cycle. The expression of Bax, Bcl-2 and caspase-3 were detected by immunohistochemistry. The proliferation of ECV-304 cells was detected by MTT assay influences. Results Compared with the control group, the ROS content of ECV-304 cells in 6.25μmol / L equol group decreased and the ROS content in 100.00μmol / L equol group increased. Compared with the control group, the levels of ROS in 25.00,50.00, The apoptosis rate of ECV-304 cells in 100.00μmol / L equol group was significantly higher than that in control group (P <0.05). The results of MTT assay showed that 0,6.25,12.50,25.00,50.00 The inhibitory rates of proliferation of ECV-304 cells in the 100.00μmol / L equol group were (8.23 ± 2.21)%, (3.29 ± 1.07)%, (6.75 ± 1.73)%, (25.46 ± 4.82)% and ± 4.66)%, low dose of equol can promote cell growth, high dose of equol has an inhibitory effect on cell growth; compared with the control group, 25.00μmol / L equol group 24 h, the intracellular Bax And caspase-3 expression increased, while Bcl-2 expression decreased. Conclusions Low concentration of equol can reduce intracellular ROS content and promote the proliferation of ECV-304 cells. High concentration of equol can lead to cell apoptosis, cell cycle arrest and inhibition of cell proliferation.