The Effects Of Vascular Endothelial Growth Factor Antisense Oligonucleotide Combined With Angiopoietin-1on Diabetic Retinopathy Rats

Objective:To observe the Leakage effect of artificial diabetic retinopathy ratsretinal neovascularization on the combination of vascular endothelial growth factorAntisense oligonucleotide(VEGF ASODN)and Angiopoietin-1(Ang-1).Method:Eighty healthy male Sprague-Dawley (SD) rats were randomly dividedinto normal control group (group A, with5rats) and the rest75rats were onceintraperitoneal injected on left lower cavity with1%concentration solution of(STZ)every60mg/Kg; After24hours, through tail vein to pick blood to measurethe blood-glucose (BG), those rats whose blood sugar value not less than16.7mmol/L is designated as diabetic rats mold. Successful diabetes rats model wereexamined by fundus fluorescein angiography (FFA)after5more months feeding,confirming the DR rats model and the rats with similar retinopathy level, randomlypicking up45rats and dividing into5groups: DR control group (Group B, with5rats);PBS buffer control group (Group C, with10rats); VEGF ASODN interventiongroup (Group D, with10rats);Ang-1intervention group (Group E, with10rats);Co-ordinate intervention group (Group F, with10rats).5μl PBS bufferintravitreal injection to C group;5μl (100μmol/L) VEGF ASODN intravitrealinjection to D group;5μl (160μg/ml) Ang-1intravitreal injection to E group; each5μl(100μmol/L) of VEGF ASODN&Ang-1(160μg/ml) intravitreal injection to F group;Then repeating above operation after3days and examining FFA, survey andcomparing the Leakage of rat's retinal neovascularization of the different groups,observing the cell nucleus of new blood vessels sprout of retina under opticalmicroscopes by histopathology specimen.Results:1.After intravitreal injection,the average Leakage area of retinalneovascularization(mm2)of Group A, B, C, D,E:0,4.53±0.71,4.46±0.62,3.14±0.22,3.34±0.26,1.53±0.24;The normal control group (group A) have no Retina leakageand neovascularization,when comparing with Group B,Group C,Group D,Group Eand Group F,the differences were significan(tP<0.01);The fluorescence leakage levelof Group B and Group C show no much differences,and it has no significance(P＞ 0.05);The fluorescence leakage level of Group D,Group E and Group F is lower thanGroup B and Group C,this differences were significant(P<0.05);The fluorescenceleakage level of Group F is lower than Group D and Group E,this differences weresignificant(P<0.05).2.Observing under optical microscope:the average number of vascular budsnucleolus over through inner limiting membrane (ILM) of Group A, B, C, D, E&Fare1.13±0.31,80.31±5.21,81.08±2.57,37.37±3.41,41.07±2.09,14.41±1.23;Group A show a nice retina condition with a similar,clear structure and neatorder,rare cases of over through ILM vascular buds nucleolus on retina,comparingGroup A with Group B,Group C,Group D,Group E and Group F,the differences wereobviously significant(P<0.01);there are many over through ILM vascular budsnucleolus are visible on the retina of Group B or Group C,the blood vessel obviouslydistended, retina structure and cell rank obviously disordered,retina tissue edema,thisdifference has no significance(P＞0.05);comparing with Group B&C,the number ofover through ILM vascular buds nucleolus of the Group D,Group E and Group F arelower,the differences were significant(P<0.05);the number of over through ILMvascular buds of The Group F is lower than Group D and Group E,the differenceswere significant(P<0.05).Conclusion:The intravitreal injection of the combaination of VEGF ASODN&Ang-1can significantly reduce retinal vessel leakage,and effectively inhibite retinaneovascularization.