Puerarin results in down-regulation of apoptosis in cultured human aortic endothelial cells induced by homocysteinemia / 中成药

ABSTRACT

AIM: To select differentially the expressed gene,glucose regulated protein 78(GRP78),to observe the expression of GRP78 in atherosclerosis tissue and to elucidate the mechanism for anti-atherosclerosis of puerarin.METHODS: The model was established by using homocysteinemia in vitro.The vascular endothelial cells were incubated with various concentrations of puerarin(10 -7 mol/L,10 -6 mol/L,10 -5 mol/L) for 48 h.The apoptosis rate was detected by flow cytometry using Annexin/PI-staining.GRP78 gene was amplificated by PCR.These differentially expressed fragments were cloned,selected,confirmed,and sequenced.Digoxigenin-labeled DNA Probe and cholesterol-rich rabit model were prepared,GRP78 mRNA was determined by in situ hybridization in atherosclerotic lesions and normal tissue.RESULTS: Three concentrtions of puerarin significantly inhibited the apoptosis of endothelial cells.The early apoptosis rate was decreased magnificently in a dose dependent manner compared with that of HCY group (P