AIM:To investigate the induction of vascular endothelial growth factor(VEGF) mRNA
expression by hypoxia in human retinal
pigment epithelial(PRE) cells,ARPE- 19 cells,and probe into the inhibitive effects of
genistein on the VEGF mRNA an d protein
expression. METHODS:The experiment was finished in the Laboratory for Physiology of Nanji ng Medical University from February to December 2004.RPE cells were exposed to h ypoxia(50 mL/L CO2 or 95 mL/L N2) for 2,12,24 and 36 hours,then the expression o f VEGF mRNA was examined by reverse transcript ploymerase chain reaction(RT- PC R).The RPE cells pretreated with 50,100,200 μ mol/L genistein and 50 μ mol/L P D98059 were exposed to hypoxia for 2 and 24 hours,then RT- PCR was used to exam ine the VEGF mRNA expression in ARPE- 19 cells,and enzyme linked immunosorbent assay(ELISA) to detect the VEGF protein expression in the culture medium. RESULTS:① Hypoxia could increase the VEGF mRNA expression in ARPE- 19 cells 2.6,3.1,8.4,2.9 times in hypoxia group as much as the normal control group at 2 ,12,24 and 36 hours(P< 0.05,n=8).② Hypoxia for 2 hours:the VEGF mRNA expression could be inhibited by 51.1% ,71.6% ,79.7% and 55% by 50,100,200 μ mol/L genistein and 50 μ mol/L PD98059 of the hypoxia group(P= 0.01,n=10),and the pro tein level in the culture medium was (97.00± 12.79),(58.85 ± 20.21),( 37.93± 14.41) and (57.83± 9.66) ng/L.③ Hypoxia for 24 hours:the VEGF mRNA exp ression could be inhibited by 55.0% ,78.7% ,90.7% and 67.2% by 50,100,200 μ mol/L genistein and 50 μ mol/L PD98059 of the hypoxia group(P=0.000,n=10),th e protein level in the culture medium was(189.60± 20.20),(117.70± 21.97),(49.7 ± 13.17) and (86.33± 12.47) ng/L. CONCLUSION:Hypoxia can induce the VEGF mRNA expression in a time- dependent manner.Genistein can suppress the upregulation of hypoxia- induced VEGF express ion in AREP- 19 cells flatly during and after RT- PCR,and inhibit the expressi on of VEGF mRNA and protein concentration- dependently.