| Objective: To detect the suppression effects of culture supernatant of human amniotic membrane on the expressions of Vascular Endothelial Growth Factor(VEGF) in rabbit comeal epithelial cells (RCEs).Methods:(1) The normal RCEs were cultured for 7 days until there were confluence on the cultures.(2) The cells were subcultured to plastic(3 groups, No.1~3,No.l was control) and naked amniotic membrane (No.4 group) cultures respectively.(3) When the cells on the plastic cultures were 70% confluence again, the RCEs were treated with DMEM (free from FBS components )medium for 12 hours.(4) Then one of plastic group (No.2 group) was cultured by culture supernatant of human amniotic membrane without epithelial layer as its medium; another plastic group (No.3 group) was cultured by culture supernatant of human amniotic membrane with epithelial layer as its medium;the other two groups(No.1 and No.4 group) were still culture by DMEM (free from FBS components ).Culturing for 48hs.(5) Then the total RNAs of these cultures were extracted and were detected by RT-PCR.Results:(l)There were expressions of VEGF mRNA from No.1 group and No.2 group, however, the expressions were significantly suppressed in No.3 group(using culture supernatant of human amniotic membrane with epithelial layer as its medium) and No.4 group (RCEs cultered directly on naked amniotic membrane) (P<0.001,n=5) .(2) The expressions of bFGF mRNA of all groups were not different significantly.Conclusion: VEGF and bFGF are important factors for corneal neovascularization,our research demonstrates that the mRNA expressions of VEGF in RCEs can be suppressed by the culture supernatant of human amniotic membrane with epithelial layer. So we think AM transplantation is partly through depressing the mRNA expression of VEGF in corneal cells to reduce the comeal neovascularization of juried comeal surface. And it may be an efficient method of curing corneal neovascularization in clinic.
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