Objective To observe the effect of adenovirus-mediated Tum5 (rAd-Tum5 )inhibiting retinal neovascularization (RNV)of oxygen-induced retinopathy (OIR)mouse model.Methods The OIR model was induced in 96 C57BL/6J mice aged of 7 days according to the literature.These mice were divided randomly into control group, OIR group, OIR rAd-green fluorescent grotein (GFP ) group and OIR rAd-Tum5 group,each group had 24 mice.The rAd-GFP and rAd-Tum5 were injected into the vitreous cavity of mice aged of 12 days in OIR rAd-GFP group and OIR rAd-Tum5 group,respectively. Meanwhile,OIR group and the control group received the injection of physiological saline solution of same volume.The relatively non-perfusion area was evaluated by fluorescence angiography,and the number of pre-retinal nucleus breaking through internal limiting membranes was observed by hematoxylin-eosin staining.The expression of vascular endothelial growth factor (VEGF)was estimated by immunofluorescent (IF)and Western blot.Results The retinal avascular areas of all groups were significantly different (F =61.224,P < 0.01 ). The retinal avascular area of the rAd-Tum5 group was decreased significantly comparing with that in the OIR group and rAd-GFP group (P <0.01 ).However,there are no significant differences between the OIR group and rAd-GFP group (P = 0.827 ).The number of pre-retinal nucleus breaking through ILM of all groups was significantly different (F =635.738,P <0.01),but no significantly difference was observed in OIR group and rAd-GFP group (P =0.261 ).Significant differences could also been seen between OIR rAd-Tum5group and OIR group as well as OIR rAd-Tum5 group and OIR rAd-GFP group (P <0.01).The results of IF and Western blot indicated that expression of VEGF in the OIR group and rAd-GFP group was obviously up-regulated, compared with that in the control group.But the expression was declined in the rAd-Tum5 group compared with that in the OIR group and rAd-GFP group. Conclusion Tum-5 peptide can efficiently prevent RNV probably by down-regulating expression of VEGF.%目的：观察腺病毒介导的重组 Tum5基因(rAd-Tum5)对氧诱导视网膜病变(OIR)模型小鼠视网膜新生血管的抑制作用。方法7日龄 C57BL/6J 小鼠96只,随机分为正常组、OIR 组、rAd-绿色荧光蛋白(GFP)空载体组(OIR rAd-GFP 组)和重组 Tum5基因组(OIR rAd-Tum5组),每组均为24只。参照文献方法建立 OIR 模型。12日龄时,OIR rAd-GFP 组、OIR rAd-Tum5组小鼠双眼玻璃体腔分别注射病毒滴度为1×1011 pfu/ml 的 rAd-GFP、rAd-Tum5病毒1μl。OIR 组、正常组小鼠双眼玻璃体腔分别注射等体积无菌生理盐水。视网膜铺片观察视网膜无灌注区相对面积,病理切片计数突破内界膜的视网膜前细胞核数,免疫荧光法及蛋白免疫印迹法检测小鼠视网膜中血管内皮生长因子(VEGF)表达情况。结果正常组、OIR 组、OIR rAd-GFP 组、OIR rAd-Tum5组小鼠视网膜无灌注区相对面积比较,差异有统计学意义(F =61.224,P <0.01)。组间视网膜无灌注区相对面积比较,OIR rAd-GFP组与 OIR 组差异无统计学意义(P =0.827)；OIR rAd-Tum5组与 OIR 组、OIR rAd-GFP 组差异均有统计学意义(P <0.01)。4组间突破内界膜的视网膜前细胞核数比较,差异有统计学意义(F =635.738,P <0.01)。组间突破内界膜的视网膜前细胞核数比较,OIR rAd-GFP 组与 OIR 组差异无统计学意义(P =0.261)；OIR rAd-Tum5组与 OIR 组、OIR rAd-GFP组差异有统计学意义(P <0.01)。与正常组小鼠视网膜 VEGF 表达比较,OIR 组和OIR rAd-GFP组 VEGF 表达均上调,OIR rAd-Tum5组小鼠视网膜 VEGF 表达较 OIR 组和 OIR rAd-GFP 组呈下降趋势。结论重组 rAd-Tum5基因可以抑制视网膜新生血管生成,可能与下调 VEGF 表达有关。
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