The Effect Of Vascular Endothelial Growth Factor And Hypoxia On The Expression Of Opiticin In Retinal Pigment Epithelium Cells.

Objective:To evaluate the role of vascular endothelial growth factor or hypoxia on the secretion of opticin in retinal pigment epithelium cells.Methods:Human RPE cells were cultured, the third to six passage of retinal pigment epithelium (RPE) cells were placed in 6-well culture plates at a density of 4×104/well. For hypoxia experiment, the cell were cultured under hypoxic condition for different times. For vascular endothelial growth factor (VEGF) experiment, the media was changed to DMEM containing different concentration VEGF for 24h respectively. Opticin and VEGF mRNA levels were determined by RT-RCR method. The protein content of opticin in RPE cells or culture media was detected by Western Blot. Matrix metalloproteinase activity in culture media was analysis by zomygraphy. One way ANOVA was used to test the comparisons between experimental groups and control group.Results:Western Blot experiment showed the opticin expression was not changed in RPE cells after hypoxia treatment, however was significantly decreased in culture media, VEGF mRNA levels in RPE cells were increased after hypoxia. Opticin expression in RPE cells was also remain unchanged after vary concentration VEGF addition treatment (F=2.29, P=0.095), and the difference in culture media was significant compared to control group (F=107.28, P<0.001). Zomygrphagy indicate a matrix metalloproteinases type 2 digest band, the activities were enhanced with VEGF increasing. The decrease of opticin in culture media after VEGF treatment could be inhibited by EDTA.Conclusions:VEGF and hypoxia have an effect on the on the secretion of opiticin in RPE cells, it may be contributed to the increasing levels of metalloproteinases type 2.