An Experimental Study Of Endostatin And Modified Endostatin And Their Angiogenesis Effect On Mouse Choroid

Objective:To observe the inhibitory effect of endostatin,poly-sulfurize heparinize endostatin(PSH-ES)and PEG-ES on choroid neovascularization and to observe the effect on expression of VEGF and PEDF,approaching the mechanism of action of endostatin and modified endostatin.To compare the difference of the inhibitory effect on choroid neovascularization among them and to find a suitable modified endostatin.Methods:80 SPF C57BL female mice were divided into 8 groups randomly,PBS group of 3d and 7d,ES group of 3d and 7d,PSH-ES group of 3d and 7d,PEG-ES group of 3d and 7d.Every group have 10 mice.All the mice were induced choroid neovascularization animal model using 532nm semiconductor laser.10μg/μl ES,PSH-ES,PEG-ES and NS 1μl were injected into vitreous cavity.HE stain and choroid surface preparation technic were used to detect the area of choroid neovascularization of every group mouse and to carry out statistics analysis.To extract choroids total RNA of every group mouse and use real-time PCR method to detect correlation factor of choroid neovascularization.To analysis VEGF and PEDF protein expression with western bloting method.Results:1.HE stain and choroid surface preparation technicEither group 3d or group 7d,compared to treated groups:the area of choroid neovascularization of control group clearly increased and the differences have statistical significance(P＜0.05).The differences between ES group and PSH-ES group or between ES group and PEG-ES group have statistical significance(P＜0.05).ES group exceeded other two groups.There are also statistical significance(P＜0.05)between PSH-ES group and PEG-ES group.2.western bloting expeimentThe results of western Noting verified that VEGF protein and PEDF protein expressed in choroids.The results of group 3d and group 7d show:the order of VEGF protein expression as follow:control group＞ES-group＞PSH-ES group＞PEG-ES group.Among the first there groups,there are statistical significance(P＜0.05)and there are no statistical significance between the last two groups(P＞0.05).The order of PEDF protein expression as follow:control group＜ES-group＜PSH-ES group＜PEG-ES group.Among the first there groups,there are statistical significance(P＜0.05)and there are no statistical significance between the last two groups(P＞0.05).3.Real-time PCRThe results of real-time PCR verified that VEGFmRNA and PEDFmRNA expressed in choroid.The results of group 3d and group 7d show:the order of VEGF mRNA expression as follow:control group＞ES-group＞PSH-ES group＞PEG-ES group.There are statistical significance among the four groups(P＜0.05).The order of PEDFmRNA expression as follow:control group＜ES-group＜PSH-ES group＜PEG-ES group.There are statistical significance among the four groups(P＜0.05).Conclusion:1.From mRNA level and protein level,we proved that endostatin,PSH-ES and PEG-ES can effectively inhibit the choroid neovascularization.2.All of ES,PSH-ES and PEG-ES can also regulate the expression of correlation factor of neovascularization.They can up-regulation the expression of PEDF and down regulation the expression of VEGF.Both PSH-ES and PEG-ES are obviously more effective than ES.3.The result show PEG-ES may be a better ES modification,thus find a new way for treatment to choroid neovascularization.