| 1. To study the expression of PCNA and FVIIIR: Ag in choroidal neovascularization (CNV), to investigate the progress and characters of CNV induced by krypton laser in BN rat.2. To study the expression and signification of PEDFmRNA, PEDPN TGFa-1 mRNA, TGFa-K TaR I in experimental CNV, and so to investigate the mechanism of PEDPN TGFa-1 in the forming progress of CNV.3. To study the inhibition of experimental CNV by rhPEDF. Methods1 . Seven groups of 42 BN rats were photocoagulated by krypton laser in single eye. 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 8 weeks and 12 weeks after photocoagulation, FFA and histopathologic examination were performed, the expression of PCNA and FVIIIR:Agwas observed by immunohistochemical method.2. Five groups of 30 rats were photocoagulated by krypton laser in single eye to induce CNV. 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks after photocoagulation, the retina was processed for histopathologic examination, in situ hybridization of PEDFmRNA and TGF- beta 1 mRNA, immunohistochemistryof PEDF, TGFa-1, TaR I and F\lR:Ag. The results of in situ hybridization and immunohistochemistry were semi-quantified, the correlation between PEDFmRNA, TGFa-1 mRNA, PEDF, TGFa-1, TaR I and FVIIIR: Ag were analysed respectively.3. Two groups of 16 BN rats were photocoagulated by krypton laser in single eye. 1 week after photocoagulation, one group of 8 rats received intravitreal injections of 2ug rhPEDF/lOfilBSS, and the other group of 8 rats received intravitreal injections of 10ul BSS as control group. 2 weeks and 4 weeks after photocoagulation, FFA was performed, and the expression of FVIII R:Ag was observed by immunohistochemical method. Results1. FFA showed fluorescein leakage 1 week after photocoagulation, and remained in 12 weeks. Subretinal new vessel was observed histopathologically 1 week after photocoagulation. The expression of PCNA decreased from 3 days to 4 weeks after photocoagulation. FVIIIR:Ag positive staining cells formed vessels in the lesion 1 week after photocoagulation; the expression of FVIIIR:Ag gradually increased and reached its peak at 4 weeks, then kept its level to 12 weeks.2. PEDFmRNA expressed in the ganglion cells, inner nuclear layer, RPE in normal BN retina. After photocoagulation, the expression of PEDFmRNA was also found in the lesions of outer nuclear layer and choroids. From 3 days to 4 weeks, the expression density of PEDFmRNA in the lesions decreased (P<0.05).The expression of PEDF was found similar to that of PEDFmRNA in normal BN retina and the retina after photocoagulation; the expression of PEDFTGF-beta1 mRNA expressed in the ganglion cells, RPE in normal BN retina and also in the lesions after photocoagulation. It was strong 3 day after photocoagulation, then became weak (P<0.05).TGF-β1 expressed in the ganglion cells, inner nuclear layer, RPE in normal BN retina. Besides that, it was found in the lesions and became weaker from 3 days to 4 weeks after photocoagulation. It concentrated at the edge of the photocoagulated lesions in 2 to 4 weeks.The expression of T/SR I was in the same spot and changed in the same way to that of TGF-β1 in normal BN retina and the retina after photocoagulation.From 3 days to 4 weeks, the expression density of FVIIIR: Ag in the lesions increased (P<0.05), however, the expression density of PEDFmRNA, PEDF, TGF-β1mRNA,TGF-βl and TβR I decreased (P<0.05); the expression of PEDFmRNA, PEDF, TGF-β1mRNA, TGF-β1> TβR I was negative correlated to that of FVIIIR:Ag during the development of CNV (r=-0.764; -0.84; -0.87; -0.87; -0.80, P<0.05) ; TGF-β was positive correlated with that of TβRI (r=0.84,P<0.05) .3. In the results of the study on the Inhibition of CNV by PEDF, 2 weeks and 4 weeks after photocoagulation, FFA showed less fluorescein leakage in the PEDF group than that in the control group, the expression of FVfflR: Ag in PEDF group was less than that in the control group at the same phase. From 2 weeks to 4 weeks after photocoagulation, the expressio... |
PDF Full Text Request