目的 探讨抗血管内皮生长因子（vascular endothelial growth factor,VEGF）药物调控线粒体功能在早产儿视网膜病变（retinopathy of prematurity,ROP）动物模型中的作用机制。方法 取18只新生（Sprague Dawley）大鼠建立氧诱导视网膜病变疾病模型,随机分为对照组（非视网膜病变）、模型组、治疗组,每组各6只。建模成功后治疗组大鼠玻璃体腔内注射1 μL抗VEGF药物（康柏西普）,其余两组大鼠玻璃体腔内注射等量生理盐水,每日1次,持续5 d。末次给药24 h后,利用荧光造影技术评估视网膜血管灌注状态并观察血管变化;苏木精-伊红（hematoxylin-eosin,HE）染色观察视网膜形态学改变;免疫组化染色检测视网膜VEGF表达;Cleaved Caspase-3免疫荧光染色评估视网膜细胞凋亡;Western blot分析视网膜中过氧化物酶体增殖物激活受体γ的辅助激活因子1-α（peroxisome proliferator-activated receptor gamma coactivator 1α,PGC-1α）、8-羟基脱氧鸟苷（8-hydroxy-2'-deoxyguanosine,8-OHdG）蛋白表达情况。结果 模型组视网膜出现大量新生血管和出血现象,而对照组与治疗组这些现象则显著减轻。与模型组比,对照组和治疗组视网膜内核层（inner nuclear layer,INL）、视网膜外核层（outer nuclear layer,ONL）的厚度均显著增大;VEGF免疫反应性减弱;Caspase-3荧光强度显著降低;PGC-1α蛋白表达水平显著升高,8-OHdG蛋白表达水平显著降低。结论 抗VEGF药物可抑制早产大鼠视网膜病变,其作用机制与调控线粒体功能并减少视网膜细胞凋亡相关。
Abstract
Objective Exploring the mechanism of anti VEGF drugs regulating mitochondrial function in an animal model of retinopathy of prematurity (ROP). Methods An oxygen induced retinal disease model was established in 18 newborn SD rats, which were randomly divided into a control group (non retinal disease), a model group, and a treatment group, with 6 rats in each group. After successful modeling, the treatment group of rats was injected with 1 μL of anti VEGF drug (Conbercept) into the vitreous cavity, while the other two groups were injected with an equal amount of physiological saline once a day for 5 days. 24 hours after the last administration, fluorescence angiography was used to observe changes in retinal blood vessels perfusion; Examine the retinal morphology via hematoxylin-eosin (HE) staining; monitor alterations in VEGF expression within the retina using immunohistochemistry; and detect changes in retinal cell apoptosis with Cleaved Caspase-3 immunofluorescence; Western blot analysis of PGC-1 α and 8-OHdG protein expression in the retina. Results The model group exhibited extensive neovascularization and retinal hemorrhaging, whereas the control and treatment groups showed a marked decrease in these occurrences compared to the model group. The inner nuclear layer (INL) and outer nuclear layer (ONL) of the retina in both the control and treatment groups showed a notable increase in thickness when contrasted with the model group; Reduced immune reactivity of VEGF; The fluorescence intensity of Caspase-3 significantly decreased; PGC-1α protein expression significantly rose, concurrently with a marked reduction in 8-OHdG protein expression. Conclusion Anti VEGF drugs can inhibit retinopathy in premature rats, and their mechanism of action is related to regulating mitochondrial function and reducing retinal cell apoptosis.
关键词
早产儿视网膜病变 /
抗VEGF药物 /
线粒体功能 /
细胞凋亡
Key words
retinopathy of prematurity /
anti-VEGF drugs /
mitochondrial function /
cell apoptosis
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基金
武汉市卫健委医学科研项目“抗VEGF药物调控线粒体功能在早产大鼠视网膜病变中的机制研究”（WX20Z25）; 湖北省卫健委医学科研项目“抗VEGF药物调控线粒体功能在早产大鼠视网膜病变中的机制研究”（2021W356）
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