Effect of Bufalin on Proliferation and Induce Apoptosis of Vascular Endothelial Cells EAhy926
|Keywords||Bufalin EAhy926 cells Proliferation Apoptosis Caspase3 Bcl-2|
Human endothelial cells were used as target cells EAhy926, through a series of research to observe bufalin as sodium pump inhibitors on vascular endothelial cell proliferation and morphological features of death. Bufalin on EAhy926 cell cycle and free Ca2+, Na+, K+ concentration change in cells were determined. Bufalin role in cells before and after the sodium pumpα1 subunit,β1 subunit, Caspase3 and Bcl-2 mRNA expression was detected by RT-PCR technique. All of the above work is foudamental to explore the bufalin in vascular endothelial cell apoptosis and death characteristics and mechanism of the process.Vascular endothelial cells EAhy926 were cultured and the effect of different concentrations of bufalin cell from 0 to 72 hours was investigated, and was found that bufalin on EAhy926 significantly inhibited cell growth depending on dose and time at range of 0.0625~1μmol/L. Apoptosis EAhy926 induced by 0.1μmol/L bufalin in 24 h was observed by fluorescence microscopic and partial cell necrosis was detected in 48 h. The typical "apoptosis peak" ranging from 8.2% to 36.5% was observed in the time-dependent manner by flow cytometry (P<0.01). Cell cycle analysis indicated that 0.1μmol/L bufalin could significantly affect the cell cycle of the vascular endothelial cells EAhy926, in which the proportion of G0/G1 phase cells was decreased during the range 0-36 h and S phase cells were significantly increased in a time-dependent changes (P<0.05). Semi-quantitative RT-PCR showed that Caspase3 mRNA expression was significantly increased compared with the control group (P<0.05), and Bcl-2 mRNA expression was markedly reduced (P<0.05), both of which showed a dose dependent manner. Confocal microscope detection showed that bufalin could increase the concentration of intracellular Na~+ and decrease the K~+ concentration in EAhy926 in a dose dependent manner (P<0.05).These results showed that bufalin inhibited EAhy926 cell proliferation, induced apoptosis, which may inhibit the sodium pump and increase the role of Caspase3 as well as decrease the Bcl-2 gene expression.