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目的:探讨血管抑素(angiostatin)玻璃体注射对氧诱导的早产儿视网膜病变(retinopathy of prematurity,ROP)的视网膜及虹膜血管渗漏的作用及其机制。方法:将出生7d(P7)的Brown Norway鼠置高氧环境(750mL/LO2)5d后再置正常氧环境诱导ROP,建立ROP动物模型,并以年龄相匹配的正常鼠作为正常对照。所有ROP鼠(P14)及正常鼠均右眼玻璃体腔注射血管抑素,左眼注射相同剂量的PBS(磷酸盐缓冲生理盐水)作为对照。用Evans蓝微血管渗透性检测法及总蛋白标准化分别于注射后1,2和3d检测视网膜和虹膜的血管渗透性;用Western blot蛋白印迹分析和免疫组化方法检测注射24h后血管内皮生长因子(vascular endothelial growth factor,VEGF)在视网膜的表达。结果:ROP鼠视网膜及虹膜的血管渗透性明显增加(P<0.01);中剂量(3.75μg/眼)和高剂量(7.5μg/眼)血管抑素降低ROP鼠视网膜血管渗透性(P<0.05,P<0.01),而低剂量组(1.88μg/眼)没有引起明显改变,呈现剂量依赖型;三种不同剂量的血管抑素玻璃体注射后ROP鼠的虹膜均未发生明显的血管渗透性的改变。血管抑素注射后第1d和第2d视网膜血管渗透性明显降低(P<0.05,P<0.01),而第3d无明显降低,其作用呈现出时间进程。Western blot蛋白印迹和免疫组化分析表明血管抑素显著降低了ROP鼠视网膜的VEGF水平,但对正常鼠无影响。结论:血管抑素可以降低ROP鼠视网膜的病理性血管渗漏,其血管渗透性下降可能与血管抑素下调VEGF的表达有关。血管抑素可能对ROP等其他视网膜血管渗漏性疾病具有潜在的治疗作用。
Objective: To investigate the effect and mechanism of vitreous injection of angiostatin on retinal and iris vascular leakage induced by oxygen in preterm infants with retinopathy of prematurity (ROP). METHODS: Brown Norway rats (7 days old) were exposed to hyperoxia (750mL / LO2) for 5 days and then exposed to normal oxygen for ROP induction. ROP animal models were established and matched with normal rats as normal controls. All ROP rats (P14) and normal mice were injected intravitreally with angiostatin in the right eye and the same dose of PBS (phosphate buffered saline) was injected into the left eye as a control. Vascular permeability of the retina and iris was detected by Evans blue microvascular permeability assay and total protein standardization at 1, 2 and 3 days after injection respectively. Western blot and immunohistochemistry were used to detect the changes of vascular endothelial growth factor vascular endothelial growth factor, VEGF) in the retina. Results: The vascular permeability of the retina and iris of ROP rats increased significantly (P <0.01). The middle dose (3.75μg / eye) and high dose (7.5μg / eye) of angiostatin decreased retinal vascular permeability , P <0.01). However, no significant changes were observed in the low-dose group (1.88 μg / eye), showing a dose-dependent manner. No significant vascular permeability was observed in iris of ROP mice after three different doses of angiostatin change. Angiostatin vascular permeability decreased significantly (P <0.05, P <0.01) on the 1st day and the 2nd day after injection of angiostatin, while no significant decrease on the 3rd day. Western blot and immunohistochemistry analysis showed that angiostatin significantly reduced the level of VEGF in the retina of ROP mice but had no effect on normal mice. Conclusion: Angiostatin can reduce the pathological vascular leakage of the retina of ROP rats, and its decreased vascular permeability may be related to the down-regulation of VEGF expression by angiostatin. Angiostatin may have a potential therapeutic effect on other retinal vascular leaks such as ROP.