玻璃体腔注射塞来昔布抑制大鼠视网膜新生血管的作用

Effect of intravitreal injection of celecoxib on neovascularization in rats

摘要:

目的：通过对大鼠氧诱导视网膜病变(oxygen-induced retinopathy，OIR)模型进行玻璃体腔注射塞来昔布，探讨塞来昔布在OIR中对视网膜新生血管的作用及其机制。

方法：选取7天龄SD乳鼠96只随机分为6组：Z组：正常组； O组：OIR组； A组：OIR+溶媒对照组； B组：OIR+塞来昔布5μg组； C组：OIR+塞来昔布20μg组； D组：OIR+塞来昔布80μg组。除Z组在正常环境饲养外，其余各组乳鼠均建立OIR模型。乳鼠于生后第 12d出氧箱后给予玻璃体内注射二甲基亚砜(dimethyl sulfoxide， DMSO)及相应剂量的塞来昔布，于生后第 17d处死，做组织切片HE染色观察视网膜组织形态并计数突破内界膜的血管内皮细胞核数，石蜡切片行免疫组化染色检测VEGF蛋白的表达。

结果：HE染色显示，Z组、O组、A组、B组、C组、D组突破视网膜内界膜的血管内皮细胞核数分别为0.44± 0.18、30.60±5.36、28.05±4.68、19.58±4.58、10.13±1.93、7.58±2.68个； 除O组与A组外，各组间差异均有统计学意义(P<0.05)，经塞来昔布治疗后，突破内界膜的血管内皮细胞核明显减少，且与剂量正相关； 免疫组化结果显示，Z组VEGF蛋白表达呈阴性，表达率为10%，其余各组可见VEGF蛋白的阳性表达，O、A组阳性表达较高，阳性率分别为90%、86%，表现为棕褐色颗粒或团块，均高于B、C、D组，且三组的阳性表达依次减低为68%、42%、30%。

结论：塞来昔布能够有效抑制大鼠OIR模型视网膜新生血管的生长，且抑制效果与剂量正相关，其作用可能通过抑制VEGF表达实现。

Abstract:

AIM: Through intravitreal injection of celecoxib in oxygen-induced retinopathy(OIR)rat model, to investigate the effect and mechanism of celecoxib on neovascularization of OIR.

METHODS: Ninety-six 7-day-old SD rats were randomly divided into 6 groups: Group Z:Normal group; Group O: OIR; Group A: OIR + vehicle control group; Group B: OIR+5μg celecoxib group; group C: OIR + 20μg celecoxib group; group D: OIR + 80μg celecoxib group. In addition to Z group in the normal environment, the other groups were established the OIR model. The neonatal rats were given intravitreal injections of dimethyl sulfoxide(DMSO)and the corresponding doses of celecoxib on the 12th day after birth. The rats were sacrificed on day 17 after birth. HE staining were employed to count the vascular endothelial cells which were breakthrough within the internal limiting membrane of retina. Immunohistochemistry staining were utilized to probe the expression of VEGF protein.

RESULTS: HE staining showed that, the number of the endothelial cells in the retina was 0.44±0.18, 30.60±5.36, 28.05±4.68, 19.58±4.58, 10.13±1.93, 7.58±2.68 in Group Z, O, A, B, C and D. In addition to Group O and Group A, there were significant differences between the two groups(P<0.05). After treatment with celecoxib, breakthrough of the internal limiting membrane of the vascular endothelial cell nucleus was significantly reduced, and positively correlated with the dose. Immunohistochemical results showed, the expression of VEGF protein in Group Z was negative, the expression rate was 10%,the positive expression of VEGF protein in Group A was higher than that in Group B, C and D, and the positive rate was 86%, which was higher than that of Group B, C and D, as 68%,42%, 30%.

CONCLUSION: Celecoxib can inhibit the OIR model of rat retinal angiogenesis, and the effect of suppressing a positive correlation with the dose,its action may inhibit VEGF expression.