Effect of hypoxia on angiogenesis ability of cardiac microvascular endothelial cell of rats

DAI Guo-hua; MA Pei-ze; SONG Xian-bo; LIU Ning; YAO Jing

doi:10.3969/j.issn.1002-266X.2014.18.004

Abstract：Objective To investigate the angiogenesis ability changes of rat cardiac microvascular endothelial cells ( CMECs) under hypoxic conditions .Methods The primarily cultured rat CMECs were cultured under hypoxia of 1%O2 , 94%N2 , and 5%CO2 for 24 h to prepare hypoxic injury models ( hypoxia group ) , meanwhile , we set the control group which was cultured under normoxic serum-free condition .Scratch healing assay was used to detect the cell migration , in-verted phase-contrast microscope was employed to count cell tube formation , MTT colorimetric assay was used to draw the cell growth curve and detect the cell proliferation rate , and dynamic observation method was applied to find the window pe-riods of cell migration , tube formation and proliferation .Real-time PCR was used to detect the expression of cell prolifera-tion-associated protein kinase (ERK) and apoptosis-related molecules (p53).Results The cell proliferation rate, migra-tion rate and tube formation rate of the hypoxia group were significantly lower than those of the control group ( all P<0.05).Compared with the control group , the angiogenesis order of the hypoxia group changed , the window period of prolif-eration became the second day in advance which was the same day of the tube formation window .The ERK mRNA relative expression levels of the hypoxia and control groups were 1.681 ±0.403 and 1.000, and p53 mRNA relative expression lev-els were 0.458 ±0.100 and 1.000.ERK mRNA expression of the hypoxia group was higher , p53 mRNA expression was lower than that of the control group (all P<0.05 ).Conclusion The angiogenesis abilities of rat CMECs significantly re-duce under hypoxic environment for 24 h, which may be related to the down-regulation of p53 gene and up-regulation of ERK gene.