急性高眼压对大鼠血—视网膜屏障的影响

发布时间：2018-06-01 15:09

本文选题：急性高眼压 + 伊文思蓝　；参考：《中南大学》2010年硕士论文

【摘要】： 目的：从视网膜的微环境入手,研究急性高眼压对大鼠BRB的影响,探讨急性高眼压后大鼠BRB损伤的时空特征以及视网膜HIF-1α的表达与BRB损伤的关系。方法：144只SD大鼠随机分为正常对照组、实验对照组和实验组。实验组大鼠制作高眼压模型,实验对照组大鼠施行假手术；之后分别存活3h、12h、Id、3d.7d。其中108只大鼠在处死前2小时大隐静脉注射3%伊文思蓝(EB),用共聚焦显微镜检测视网膜铺片和切片中EB的分布；用分光光度计定量视网膜EB的含量。36只大鼠用免疫组化方法检测视网膜HIF-1α的表达。结果：在正常视网膜中偶见EB红色荧光斑点,在急性高眼压后视网膜中可见EB红色荧光斑点主要分布在节细胞层的毛细血管周边,3h、12h和1d时EB红色荧光斑点还见于内核层的毛细血管周边。视网膜中央部在3h时EB红色荧光斑点开始增多,12h时EB红色荧光斑点最多,然后逐渐减少；视网膜周围部在3h时EB红色荧光斑点最多,然后逐渐减少；视网膜周围部先于中央部出现EB渗漏高峰。进一步定量检测显示和正常对照组相比,视网膜中央部在3h时EB渗漏量开始增多,12h时EB渗漏量最多,以后逐渐减少,7d时接近恢复正常；视网膜周围部在3h时EB渗漏量最多,以后逐渐减少,7d时接近恢复正常；视网膜周围部先于中央部出现EB渗漏量峰值。免疫组化结果显示视网膜HIF-1α在正常对照组未见阳性表达,在HIOP后3h可见HIF-1α阳性产物增多,12h时达高峰,然后1d、3d、7d逐渐减少；从视网膜中央部和周围部HIF-1α表达情况看,在3h和1d时视网膜周围部HIF-1α阳性产物明显比中央部多；HIF-1α阳性产物主要见于节细胞层,3h、12h时HIF-1α阳性产物还见于内核层。结论：1.大鼠BRB在急性高眼压后的早期损伤更明显,晚期逐渐恢复；视网膜周围部BRB先于中央部出现严重破坏；BRB损伤主要分布于节细胞层。 2.急性高眼压后HIF-1α的表达在早期明显上调,晚期逐渐恢复,其表达的时空特征与BRB损伤的时空特征一致,提示急性高眼压后HIF-1α的表达上调可能与BRB的损伤有关。
[Abstract]:Aim: to study the effect of acute intraocular pressure (IOP) on BRB in rats from the microenvironment of retina, and to investigate the temporal and spatial characteristics of BRB damage in rats with acute intraocular hypertension and the relationship between the expression of HIF-1 伪 in retina and BRB damage. Methods 144 Sprague-Dawley rats were randomly divided into normal control group, experimental control group and experimental group. The rats in the experimental group were made into the model of high intraocular pressure, and the rats in the control group were sham-operated, and then survived for 3 hours and 12 hours, respectively. 108 rats were injected with 3% Evans blue EBV 2 hours before death. The distribution of EB in retinal slices and sections was detected by confocal microscope. The content of EB in retina was measured by spectrophotometer. The expression of HIF-1 伪 in retina of 36 rats was detected by immunohistochemical method. Results: EB red fluorescent spots were occasionally seen in the normal retina. EB red fluorescent spots were found in the retina after acute intraocular pressure (IOP), mainly in the capillary periphery of the ganglion cell layer at 12 h and 1 d after intraocular pressure, and also around the capillary in the nuclear layer. The number of EB red fluorescent spots in the central part of the retina began to increase at 3 h and then decreased gradually at 12 h, and then decreased gradually at 3 h in the peripheral retina, the number of EB red fluorescent spots in the central part of the retina increased at 3 h and then decreased gradually at 12 h. The peak of EB leakage occurred in the periretinal region before the central part. Further quantitative examination showed that EB leakage in the central part of the retina began to increase at 3 h and reached the maximum at 12 h, then decreased gradually at 7 d, and returned to normal at 7 d, and that in the peripheral retina at 3 h, the EB leakage was the highest in the central part of the retina compared with the normal control group. After 7 days of gradual reduction, the peak of EB leakage occurred in the peripheral part of the retina before that in the central part. Immunohistochemical results showed that there was no positive expression of HIF-1 伪 in the normal control group, and the positive product of HIF-1 伪 reached its peak at 12 h after HIOP, and then decreased gradually at 1 d 3 d and 7 d, and the expression of HIF-1 伪 was observed in the central and peripheral retina. At 3 h and 1 d, the positive products of HIF-1 伪 in the periretinal region were significantly higher than those in the central part of the retina. The positive products of HIF-1 伪 were mainly found in the ganglion cell layer at 3h and 12h, and also in the nuclear layer. Conclusion 1. The early injury of BRB was more obvious after acute high intraocular pressure, and the late stage gradually recovered, and the severe damage of BRB in the periretinal region was mainly distributed in the ganglion cell layer before the central part of the retina. 2. The expression of HIF-1 伪 was obviously up-regulated at the early stage and gradually recovered at the late stage after acute intraocular hypertension. The spatiotemporal characteristics of HIF-1 伪 expression were consistent with those of BRB injury, suggesting that the up-regulation of HIF-1 伪 expression after acute intraocular hypertension might be related to BRB damage.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R774.1

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