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Published on: March 18, 2015
[Effect of PAI-1 antisense RNA on vascular endothelial growth factor expression in aorta smooth muscle cells cultured
1Department of Cardiology of the First Affiliated Hospital, Harbin Medical University, Harbin, Heilongjiang 150001 P. R. China. xinneiyi@0451.com
Objective:
To investigate the role of plasminogen activator inhibitor-1(PAI-1) antisense RNA in regulating the expression of PAI-1 and vascular endothelial growth factor (VEGF) in aorta smooth muscle cells cultured in vitro.
Methods:
The second extron of PAI-1 was amplified with polymerase chain reaction(PCR), the production was inserted into eukaryotic cell expression vector pcDNA3.1 after it had been purified and cloned so as to construct PAI-1 antisense RNA recombination plasmid. The recombination plasmid was transfected into SMC. PAI-1 expression was detected by immunohistochemistry, Western blod and ELISA; the effects of PAI-1 variation on VEGF was examined by immunofluorescence.
Results:
PAI-1 antigen was the lowest in cells on the third day after transfection; the expression of VEGF was also decreased. PAI-1 antigen gradually increased on the fifth day and VEGF increased correspondingly. On the seventh day, PAI-1 antigen and VEGF increased to nearly normal level.
Conclusion:
PAI-1 antisense RNA can block the translation progress of PAI-1 proteins effectively and inhibit the expression of VEGF in aorta smooth muscle cells.
