Objective: To investigate the effect of antisense KLF4 (Krüppel-like factor 4) gene on the expression of vWF and PAI-1 in endothelial cells.
Methods: Human umbilical vein endothelial cells (HUVEC) were isolated from umbilical vein and cultured in endothelial cell medium. The recombinant adenoviral plasmid carrying the antisense KLF4 gene was constructed by homologous recombination. KLF4, PAI-1 and vWF mRNAs and proteins expression were detected by real time-PCR, Western blot, and confocal laser microscopy.
Results: Recombinant adenoviral plasmid carrying the antisense KLF4 gene (Ad-KLF4AS) was constructed successfully. Compared with the control Ad-GFP infection group, Ad-KLF4AS at a 200 MOI can down-regulate the expression of KLF4 gene in HUVEC (from 0.59 ± 0.01 to 0.44 ± 0.06) (P < 0.05), and increase vWF mRNA (from 1.04 ± 0.03 to 1.17 ± 0.05) and protein expression (P < 0.05). PAI-1 mRNA and protein of Ad-KLF4AS infection group was higher than that of Ad-GFP infection group. PAI-1 mRNA between the two groups had no significant difference (P > 0.05).
Conclusions: Down-regulation of KLF4 leads to increase in expression of vWF and PAI-1 in endothelial cells. KLF4 might play an important role in regulation of endothelial coagulant function.