曲安奈德抑制氧诱导视网膜病变模型小鼠中视网膜新生血管的机制研究

发布时间：2018-04-24 03:30

本文选题：曲安奈德 + 血管内皮细胞生长因子　；参考：《复旦大学》2011年硕士论文

【摘要】：第一部分氧诱导视网膜病变(oxygen-induced retinopathy, OIR)模型的建立目的建立OIR模型小鼠。方法7日龄C57BL/6J新生小鼠6只作为单纯给氧组,置于氧浓度75%的容器内连续饲养5天至小鼠12日龄,再置于正常空气环境下饲养至17日龄；6只同龄新生小鼠在正常空气环境饲养至17日龄作为单纯对照组。采用FFA眼底造影、视网膜铺片ADP染色及计数眼球切片中突破视网膜内界膜新生血管的内皮细胞核数等方法观察视网膜新生血管情况。结果单纯给氧组FFA检查可见眼底视网膜新生血管生成,视网膜铺片ADP酶染色见视网膜血管大片无灌注区,血管部分闭塞,分支减少,分布较紊乱；而单纯对照组小鼠视网膜血管分布均匀规则,无闭塞。单纯给氧组平均每个眼球切片中突破视网膜内界膜的内皮细胞核数为(24.8±7.1)个,较对照组显著增多(P0.05)。结论该小鼠模型具有血管无灌注区、血管闭塞、新生血管形成等改变,重复性好、可定量研究,是研究视网膜新生血管比较合适的动物模型。第二部分曲安奈德(triamcinolone acetonide, TA)抑制视网膜新生血管的机制研究目的通过OIR模型小鼠,探讨TA抑制视网膜新生血管的可能机制。方法7日龄C57BL/6J新生小鼠36只,共分为4组。其中18只制作OIR动物模型,并且于12日龄时玻璃体腔内注射曲安奈德2μl,作为高氧TA组；对侧眼注射相同体积的平衡盐溶液(balanced salt solution, BSS),作为高氧BSS组；其余两组小鼠在正常环境下饲养,余处理同前,分别作为正常BSS组和正常TA组。用FFA造影及二磷酸腺苷(adenosine diphos-phate, ADP)酶法染色,了解视网膜血管情况；用视网膜切片HE染色,计数小鼠视网膜新生血管内皮细胞核数,加以比较；用免疫组织化学法检测各组小鼠视网膜切片VEGF、SDF-1、CD14含量。用Realtime PCR检测VEGF、SDF-1 mRNA的表达。结果与高氧BSS组相比,高氧TA组突破视网膜内界膜内皮细胞细胞核数目明显减少(P0.05),血管分布更规则,FFA造影接近正常眼底。SDF-1、VEGF mRNA及SDF-1、VEGF和CD14蛋白表达在高氧TA组低于高氧BSS组(P0.05),正常TA组和正常BSS组之间无明显差异(P0.05)。VEGF、SDF-1与CD14的IOD/AOI值均呈显著线性相关(偏相关分析P0.05)。结论TA能有效抑制小鼠视网膜新生血管形成,其机制可能是通过减少单核/巨噬细胞聚集进而降低SDF-1及VEGF的表达而实现。
[Abstract]:The first part is the establishment of oxygen-induced retinopathy (OIRs) model. Objective to establish OIR model mice. Methods six 7-day-old C57BL/6J newborn mice were fed in a 75% oxygen concentration container for 5 days to 12 days of age. Six newborn mice of the same age were reared to 17 days of age in normal air as the control group. The retinal neovascularization was observed by FFA fundus angiography, ADP staining and counting the number of endothelial nuclei of neovascularization through the inner limiting membrane of the retina. Results the retinal neovascularization was observed in the oxygenated group by FFA. The retinal ADP enzyme staining showed that there was no perfusion area, partial occlusion, less branches and more disordered distribution of retinal vessels in the oxygenated group. In the control group, the retinal blood vessels were distributed uniformly and without occlusion. The number of endothelial nuclei breaking through the inner retinal membrane was 24.8 卤7.1 in the oxygen alone group, which was significantly higher than that in the control group (P 0.05). Conclusion the mouse model has no perfusion area, vascular occlusion, angiogenesis and so on. It has good reproducibility and can be quantitatively studied. It is a suitable animal model for the study of retinal neovascularization. The mechanism of triamcinolone acetonide (TA) in inhibiting retinal neovascularization Objective to explore the possible mechanism of TA inhibiting retinal neovascularization in OIR mice. Methods 36 7-day-old C57BL/6J mice were divided into 4 groups. Eighteen OIR animal models were made and triamcinolone acetonide (2 渭 l) was injected intravitreally as hyperoxia TA group at 12 days of age, and balanced salt solution in the same volume was injected into the contralateral eye as hyperoxia BSS group. The other two groups of mice were fed in normal environment. The remaining mice were treated as normal BSS group and normal TA group respectively. The retinal vessels were studied by FFA and adenosine diphos-phosphate enzyme staining, and the number of endothelial nuclei of retinal neovascularization in mice were counted by HE staining in retina sections, and the number of endothelial nuclei of retinal neovascularization in mice was compared. Immunohistochemical method was used to detect the content of CD14 in retina sections of mice. The expression of VEGFG SDF-1 mRNA was detected by Realtime PCR. Results compared with hyperoxia BSS group, In hyperoxia TA group, the number of endothelial cell nucleus of retinal inner boundary was significantly reduced, and the expression of VEGF mRNA and SDF-1 mRNA and CD14 protein in hyperoxia TA group was significantly lower than that in hyperoxia TA group compared with that in hyperoxia BSS group, while in normal TA group and positive group, the expression of VEGF-VEGF and CD14 in hyperoxia TA group was more regular than that in hyperoxia TA group. There was no significant difference between normal BSS group and normal BSS group. There was a significant linear correlation between IOD/AOI value of CD14 and SDF-1 (partial correlation analysis P 0.05). Conclusion TA can effectively inhibit the formation of retinal neovascularization in mice, which may be achieved by reducing mononuclear / macrophage aggregation and thus the expression of SDF-1 and VEGF.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1

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