Role Of Tyrosine Kinase Inhibitor (Genistein) In Retinal Neovascularization In Mice

Objective: To investigate the effect of tyrosinase inhibitor(Genistein)on retinal neovascularization in mice.Methods: Using random number table method,36 7-day-old clean C57BL/6J mice were randomly divided into model control group,Gen group,DMSO group,normal group.Each group had 9 mice.The first three groups were returned to normal environment after 5 days in the closed oxygen box with oxygen volume fraction(75 ± 2)%.The normal group was the mice in the natural environment.The model control group and normal group were not given drug treatment.Gen group and DMSO group of mice for 12 days old,intravitreal injection of Genistein(diluted with 5% dimethyl sulfoxide(DMSO),400mg/L)and DMSO 1?l respectively.The mice were cultured under normal conditions for 5 days.At seventeenth days after birth,2 mice in each group were subjected to fluorescein angiography to observe the morphology of the retinal vessels.All the remaining mice were sacrificed to obtain the retina.Real-time PCR were used to detect the expression of VEGF and b FGFm RNA in the mouse retina(2-??Ct),and the expression of VEGF and b FGF proteins in the retina of mice were detected by Western blot(VEGF,b FGF/?-actin).Results: The m RNA expression levels of VEGF and b FGF in each group were as following:(0.64±0.25 and 21.40±3.07)in model control group,(0.37±0.23 and 17.22 ± 2.63)in DMSO group,(0.03 ± 0.02 and 0.52 ± 0.25)in Gen group and(0.04 ± 0.02 and 0.67 ± 0.23) in the normal group,and model control group and DMSO group were significantly higher than the normal group and Gen group.By the pairwise comparisons,the m RNA expression levels of VEGF and b FGF between the model control group and the DMSO group,normal group and Gen group were not statistically significant(P>0.05).By the other pairwise comparisons,the differences were statistically significant(P<0.05).The protein expression levels of VEGF and b FGF in each group were as following:(1.01 ± 0.05 and 0.97 ±0.06)in model control group,(1.06±0.07 and 1.03 ± 0.08)in DMSO group,(0.73 ± 0.05 and 0.76 ± 0.07)in Gen group and(0.52 ± 0.05 and 0.56 ± 0.05)in the normal group,and model control group and DMSO group were significantly higher than the normal group and Gen group.The protein expression levels of VEGF and b FGF between model control group and DMSO group had no statistical difference(P>0.05).By the other pairwise comparisons,the differences were statistically significant(P<0.05).Fluorescein angiography of retinal: In the model control group and DMSO group,there were many layers and branches of blood vessels and new vascular clusters.In group Gen,the retinal vessels were stratified and the branches were less.The retinal vessels were relatively clear,and the number of new vessels was less.The normal group retinal vessel shape neat,no neovascularization clusters.Conclusion: 1.High expression of VEGF and b FGF in oxygen induced retinal neovascularization in mice;2.Genistein can inhibit the formation of VEGF and b FGF in retinal neovascularization;3.Genistein can inhibit the formation of retinal neovascularization.