目的观察卷柏总黄酮、穗花杉双黄酮对人脐静脉内皮细胞(HUVEC)增殖及血管内皮细胞生长因子(VEGF)表达的影响。方法采用四甲基偶氮唑蓝(MTT)法检测不同浓度卷柏总黄酮及穗花杉双黄酮对HUVEC细胞增殖的影响;流式细胞仪检测细胞周期;Western blot检测VEGF蛋白的表达。结果不同质量浓度的卷柏总黄酮(1、5、10、15、20μg.mL-1)和穗花杉双黄酮(0.3,1,2,4,5μg.mL-1)可以剂量依赖地促进HUVEC细胞增殖,其中卷柏总黄酮10μg.mL-1、穗花杉双黄酮1μg.mL-1在36 h时显著促进细胞增殖,提高S期细胞比例,VEGF蛋白表达明显升高。结论细胞实验结果提示卷柏中黄酮类物质可能具有促血管新生的作用。
OBJECTIVE To observe the effects of total flavonoids and amntoflavone isolated from Selaginella tamariscina on the cell proliferation and cycle and the expression of vascular endothelial growth factor(VEGF) in human umbilical vein endothelial cells(HUVEC).METHODS HUVEC was cultured as target cell in vitro.MTT method was used for the analysis of the effect on the proliferation of HUVEC by different concentrations of total flavonoids and amntoflavone isolated from Selaginella tamariscina;flow cytometer was used to determine the cell cycle,and Western blot was wtilized to investigate the expression of VEGF protein.RESULTS After 24 h treatment total flavonoids(1,5,10,15,20 μg·mL-1) and amntoflavone(0.3,1,2,4,5 μg·mL-1) isolated from Selaginella tamariscina.Promoted the proliferation of HUVEC with remarkable dosage-effect relationship,and the total flavonoids and amntoflavone had the best proliferation effect at the concentrations of 10 and 1 μg·mL-1,respectivey.Further studies showed that total flavonoids(10 μg·mL-1) and amntoflavone(1 μg·mL-1) could both significantly enhance the synthesis of DNA and increase VEGF protein expression of HUVEC in 36 h.CONCLUSION Experimental results demonstrate that the total flavonoids and amntoflavone isolated from Selaginella tamariscina may have the effect of inducing angiogenesis.