| Objective:To study the effect of Heterophyllin B(HB),a water-soluble active component of Radix Pseudostellariae,on angiogenesis and its related mechanism,and to provide reference for clinical research and application of Radix Pseudostellariae.Method:Chicken embryo chorioallantoic membrane(CAM)experiment was used to investigate the effect of HB on CAM angiogenesis;Use CCK8 assay to detect the effect of HB on Human umbilical vein endothelial(HUVEC)cells proliferation;scratch assay was used to detect the effect of HB on HUVEC migration;ELISA was used to measure the secretion level of Vascular endothelial growth factor165(VEGF165)and fibroblast growth factors2 of HB-treated HUVEC;Western Blot was applied to analyze the possible mechanisms by which HB promotes angiogenesis.Result:The results of CAM experiments showed that HB had a significant effect on angiogenesis.CCK8 results suggested that HB could promote the proliferation of HUVEC in the concentration range of 40 to 120μM.Scratch assay showed that there was a significant difference in healing degree at different HB concentrations groups when compared with control(P < 0.05).With the increase of HB concentration,the migration intensity of HUVEC was enhanced,which was time-and concentration-dependent.ELISA indicated that the up-regulation of expression of VEGF165 was positively correlated with HB concentration(P < 0.05).Compared with HB group,the expression of VEGF165 was significantly different in negative and positive control groups(P < 0.05).Western Blot showed that the expression of KRAS,RAF-1,MEK1/2 and ERK1/2 were positively correlated with HB concentration(P < 0.05).Conclusion:HB can promote angiogenesis,which may be related to up-regulation the expression of VEGF165 in HUVEC,activation of MAPK downstream ERK signal pathway,and promotion of proliferation and migration of HUVEC cells. |
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