穗花杉双黄酮抑制小儿血管瘤内皮细胞体外增殖实验研究

Inhibitory effect of Amentoflame on proliferation of endothelial cells from infantile hemangioma in vitro

目的:探究穗花杉双黄酮(AF)对小儿血管瘤内皮细胞体外增殖的影响。方法:MTS法筛选AF抑制小儿血管瘤内皮细胞体外增殖的适宜浓度,流式细胞法检测AF对小儿血管瘤内皮细胞细胞周期G 1期和S期分布的影响,荧光定量PCR和Western blot法检测AF对小儿血管瘤内皮细胞增殖相关蛋白表达的影响。结果:75、125μmol/L AF显著降低小儿血管瘤内皮细胞活性(P<0.05),75μmol/L AF用于后续细胞周期实验、荧光定量PCR和Western blot实验,结果显示75μmol/L AF抑制小儿血管瘤内皮细胞从G 1期进入S期(P<0.05),但对G 2其细胞分布无明显作用,与对照组相比较,AF显著抑制小儿血管瘤内皮细胞β-catenin和Cyclin D1 mRNA和蛋白表达,促进p21 mRNA和蛋白表达(均P<0.05)。结论:AF通过抑制β-catenin和Cyclin D1,促进p21等增殖相关蛋白表达进而抑制小儿血管瘤内皮细胞体外增殖。

Objective:To explore the effect of Amentoflame(AF)on the proliferation of endothelial cells from infantile hemangioma in vitro.Methods:MTS method was used to screen the appropriate concentration of AF to inhibit the proliferation of endothelial cells from infantile hemangioma in vitro.Flow cytometry was used to detect the effect of AF on the distribution of endothelial cells from infantile hemangioma in the G 1 and S phases.Fluorescence quantitative PCR and Western blot were used to detect the effect of AF on the expression of proliferation-related proteins in the endothelial cells from infantile hemangioma.Results:75 and 125μmol/L AF significantly reduced the activity of endothelial cells from infantile hemangioma.75μmol/L AF was used in subsequent cell cycle experiments,fluorescent quantitative PCR and Western blot experiments.The results indicated that 75μmol/L AF inhibited the endothelial cells from infantile hemangioma to enter the S phase from G 1 phase,but had no obvious effect on the cell distribution of G 2 phase.Compared with the control group,AF significantly inhibited the expression ofβ-catenin and Cyclin D1 mRNA and protein in endothelial cells from infantile hemangioma,and promoted the expression of p21 mRNA and protein(all P<0.05).Conclusion:AF can promote the expression of proliferation-related proteins such as p21 by inhibitingβ-catenin and Cyclin D1,and thereby can inhibit the proliferation of endothelial cells from infantile hemangioma in vitro.