Xuefu Zhuyu Decoction Induces Endothelial Cells Angiogenic Activity And Its Mechanisms

Objective To investigate the effect of Xuefu Zhuyu Decoction induce endothelial cells angiogenic activity and its mechanisms.Methods Serum pharmacology technique was adopted.Human umbilical vein endothelial cells(HUVECs) were incubated with blank serum,Xuefu Zhuyu Decoction containing serum at final concentrations of 1.25%,2.5%and 5%.HUVECs migration,invasion,cell viability, numbers of cells in S phase,adhesion and capillary tube formation were assessed by wound healing assay,boyden chamber invasion assay,Methyl thiazolyl tetrazolium(MTT) assay,cell cycle assay,gelatin adhesion assay and capillary tube formation assay respectively.Secretion of vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) were evaluated by enzyme-linked immunosorbent assay(ELISA);secretion of nitric oxide(NO) was determined by nitrate reductase;synthesis of NO was studied by fluorescent probe; endothelial nitric oxide synthase(eNOS) activity was assessed by enzymatic colorimetry; VEGF,bFGF and eNOS mRNA level were evaluated by Reverse Transcriptase Polymerase Chain Reaction(RT-PCR).Results When HUVECs were incubated with Xuefu Zhuyu Decoction containing serum for 24h,the resulsts showed that final concentrations of 1.25%,2.5%and 5%upregulated bFGF mRNA level,secretion of bFGF and induced HUVECs migration.When HUVECs were incubated with Xuefu Zhuyu Decoction containing serum for 48h,the resulsts showed that final concentrations of 1.25%induced eNOS mRNA level,NO secretion,synthesis of NO, activity of eNOS,HUVECs capillary tube formation;2.5%enhanced VEGF,eNOS mRNA level,VEGF and NO secretion,synthesis of NO,activity of eNOS,HUVECs migration,invasion,cell viability,numbers of cells in S phase,adhesion and capillary tube formation;5%stimulated VEGF,eNOS mRNA level,bFGF and NO secretion,synthesis of NO,activity of eNOS,HUVECs migration,invasion,adhesion and capillary tube formation.Conclusions Xuefu Zhuyu Decoction induces HUVECs angiogenesis by upregulating the mRNA and protein level of VEGF,bFGF and eNOS,synthesis and secretion of NO.This suggests that the mechanism of Xuefu Zhuyu Decoction enhancing angiogenesis for treating ischemic diseases may be accounted for these effects.