The Study Of The Effect Of CD105(Endglin) On Corneal Vascularization

Objective To establish an animal model of corneal neovascularization(CNV) and tosupply a steady condition for the investigation of CNV. To study the expression ofCD105 in corneal neovascularization (CNV) of rats, and study its relation with VEGF. Toinvestigate the effect of silencing CD105 gene expression with siRNA on theproliferation of Human umbilical vein endothelial cells (HUVECs) and VEGF mRNAand protein expression.Methods The pills containing 100 ng/μl the rat-derived basic fibroblast growth factor(b-FGF) 1μl were transplanted into the corneal pockets of rats to establish the rat CNVmodels. At the indicated time points, morphologic changes in CNV were observed, andthe expression of CD 105 and VEGF was detected by immunohistochemistry, computerimage analysis and RT-PCR. Human umbilical vein endothelial cells were cultured invitro and transfected with siRNA of CD105 by lipofectamine2000. The levels of CD105mRNA and VEGF mRNA expression before and after transfection were detected bysemi-quantitative reverse transcription polymerase chain reaction (RT-PCR). The levelsof CD105 protein and VEGF protein expression before and after transfection weredetected by western-blot. Apoptosis of HUVECs induced by CD105 siRNA was detectedby MTT.Results The CNV began to grow on the 4th day, reached the peak on the 10th day,and reduced after 3 weeks. The expression of CD105 and VEGF was parallel to the growth of CNV. In vitro, The RT-PCR detected that the level of CD105 mRNA inHUVECs was inhibited by the special siRNA. Western-blot indicated that the expressionof CD105 protein was downregulated in HUVECs after interfered with siRNA. WhileVEGF mRNA and protein were upregulated parallel with that of CD105. The cellproliferation in RNAi group was significantly lower than that in the negative control andcontrol.Conclusion The bFGF pills could induce the CNV in rats' cornea successfully. Thegel foam sponges-bFGF-2％agarose inducing CNV in rat eye was feasibility and idealanimal model because of its high CNV formation rate, repetitive and easy for observationand measurement in vivo. CD105 has a relation to the CNV and plays a necessary role inthe growth of CNV. RNA interference can downregulate the level of CD105 mRNA andCD105 protein expression, through that VEGF mRNA and protein can be downregulatedalso, then induce the apoptosis of HUVECs. CD105 can accelerate the growth ofHUVECs. CD105 may be used as a gene therapy target on corneal neovascularization.