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α-黑素细胞刺激素在氧诱导的视网膜病变小鼠模型中抑制病理性视网膜新生血管

发表时间:2016-03-15  浏览量:1696  下载量:548
全部作者: 杨千惠,张琰,薄其玉,曹云山,杨伟,李筱荣
作者单位: 天津医科大学眼科医院,天津医科大学眼科研究所,天津医科大学眼视光学院;甘肃省人民医院心血管内科
摘 要: 目的:以氧诱导视网膜病变(oxygen-induced retinopathy,OIR)小鼠为模型,研究玻璃体腔注射不同浓度的α-黑素细胞刺激素(α-melanocyte-stimulating hormone,α-MSH)对病理性视网膜新生血管(retinal neovascularization,RNV)生成的作用。方法:随机选取健康清洁级C57BL/6J幼鼠(7日龄)分为OIR+α-MSH-1组、OIR+α-MSH-5组、OIR+α-MSH-10组、OIR组和正常对照组。不同剂量α-MSH干预组和OIR组幼鼠在高氧环境下[氧浓度(75±2)%] 饲养5 d,然后在普通环境中饲养5 d,正常对照组幼鼠则在普通空气环境下饲养10 d. 取17日龄鼠,球后静脉注射高分子量异硫氰酸葡聚糖(fluorescein isothiocyanate-dextran,FITC-dextran),制备视网膜铺片,观察视网膜的血管形态,计算无灌注区面积相对于整个视网膜的面积;对小鼠眼球行石蜡切片和苏木精-伊红(hematoxylin-eosin,HE)染色,计数突破内界膜突入玻璃体腔的血管细胞核数。结果:视网膜铺片结果显示,各组间视网膜无灌注区相对面积整体比较有统计学意义(F=19.64,P<0.05)。其中正常对照组未见无灌注区,OIR组无灌注区的相对面积为(23.01±3.39)%,二者间差异有统计学意义(P<0.001)。而α-MSH干预组无灌注区的相对面积为OIR+α-MSH-1组,(18.14±7.20)%;OIR+α-MSH-5组,(15.64±7.07)%;OIR+α-MSH-10组,(7.62±6.52)%. 其中,OIR+α-MSH-10组与OIR组相比,无灌注区的相对面积有统计学差异(P<0.01)。石蜡切片HE染色结果表明,OIR组平均每张切片突入玻璃体腔的视网膜血管细胞核数为(13.59±4.28)个,而正常组为(1.48±1.27)个,二者差异有统计学意义(P<0.001)。α-MSH各干预组平均每张切片突入玻璃体腔的血管细胞核数分别为OIR+α-MSH-1组,(6.35±2.34)个;OIR+α-MSH-5组,(4.96±1.79)个;OIR+α-MSH-10组,(1.03±1.25)个。α-MSH干预组比较OIR组血管细胞核数明显减少(P<0.01),且实验组间比较有统计学意义(F=147.87,P<0.05)。结论:α-MSH在OIR小鼠模型中,剂量依赖性地减少视网膜中无灌注区的相对面积和视网膜新生血管核数,对病理性RNV生成有抑制作用。
关 键 词: 眼科学;氧诱导视网膜病变;视网膜新生血管;α-黑素细胞刺激素;病理
Title: α-melanocyte-stimulating hormone inhibits pathological retinal neovascularization in a mouse model of oxygen-induced retinopathy
Author: YANG Qianhui, ZHANG Yan, BO Qiyu, CAO Yunshan, YANG Wei, LI Xiaorong
Organization: School of Optometry and Ophthalmology, Tianjin Medical University, Tianjin Medical University Eye Institute, Tianjin Medical University Eye Hospital; Department of Cardiology, Gansu Provincal Hospital
Abstract: Objective: To study the effects of intravitreal injection of α-melanocyte-stimulating hormone (α-MSH) at different concentrations on pathological retinal neovascularization (RNV) in a mouse of oxygen-induced retinopathy (OIR). Methods: Healthy C57BL/6J pups of hygiene grade were randomly divided into OIR+α-MSH-1, OIR+α-MSH-5, OIR+α-MSH-10, OIR and normal control groups at postnatal day 7 (P7). The α-MSH intervention groups and OIR group were exposed to high oxygen [(75±2)%] for 5 d, then maintained under normal air condition for another 5 d; whereas the normal control group was raised under normoxia for 10 d. Retro-orbital injection of high molecular weight fluorescein isothiocyanate-dextran (FITC-dextran) was performed on P17 mice. The retina whole mounts were prepared to observe retinal vessels and quantify the relative area of vessel obliteration. The mouse eyeballs were subjected to paraffin sections and hematoxylin-eosin (HE) staining, and the average number of pre-retinal nuclei per section was quantified. Results: FITC-dextran-labeled retinal whole mounts showed that the relative vessel obliteration area was of statistical significance among the groups (F=19.64, P<0.05). No vessel obliteration was observed in normal control retinas, whereas the relative avascular area in the OIR group was (23.01±3.39)%, the difference between the two groups was of statistical significance (P<0.001). The relative avascular area in OIR+α-MSH-1, OIR+α-MSH-5, and OIR+α-MSH-10 groups were (18.14±7.20)%, (15.64±7.07)%, and (7.62±6.52)%, respectively. The relative avascular area in the OIR+α-MSH-10 group was significantly lower than that in the OIR group (P<0.01). HE staining of the paraffin sections showed that the average number of pre-retinal nuclei per section was 13.59±4.28 in the OIR group, and 1.48±1.27 in the normal controls, there is statistical significance between the two groups (P<0.001). The average numbers of pre-retinal nuclei per section in the α-MSH-treated groups were 6.35±2.34 for OIR+α-MSH-1 group, 4.96±1.79 for OIR+α-MSH-5 group, and 1.03±1.25 for OIR+α-MSH-10 group, and these numbers were all significantly less than that in the OIR group (P<0.001) and different compared with each group (F=147.87, P<0.05). Conclusion: Intravitreal injection of α-MSH dose-dependently reduces the relative area of vessel obliteration and the average number of pre-retinal nuclei in the retinas of OIR mouse model, thereby inhibiting RNV under the pathological condition.
Key words: ophthalmology; oxygen-induced retinopathy; retinal neovascularization; α-melanocyte-stimulating hormone; pathology
发表期数: 2016年3月第5期
引用格式: 杨千惠,张琰,薄其玉,等. α-黑素细胞刺激素在氧诱导的视网膜病变小鼠模型中抑制病理性视网膜新生血管[J]. 中国科技论文在线精品论文,2016,9(5):435-442.
 
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