大鼠急性视网膜缺血节细胞死亡时脑红蛋白的表达  

The neuroglobin expression when retinal ganglion cells death in acute rats＇ retina ischemia

作　　者：丁国鹏[1,2] 燕振国[3] 何涛[1] 游思维[3] 李耀宇[1] 沈学峰[3] 王涛[3] 张欲凯[3] 黄晓峰[4] 

机构地区：[1]北京军区总医院眼科,100700 [2]山西医科大学第二临床医院 [3]第四军医大学全军神经科学研究所,西安 [4]第四军医大学电镜中心,西安

出　　处：《中华眼科杂志》2010年第7期590-596,共7页Chinese Journal of Ophthalmology

摘　　要：目的 研究大鼠视网膜急性缺血中视网膜节细胞（RGCs）死亡时脑红蛋白（Nsb）的表达.方法 实验研究.采用特异性结扎大鼠视网膜动脉的方法造成视网膜急性缺血模型,按照结扎时间不同（0、15、30、60 min）分为A、B、C、D组,每组10只大鼠,均以左眼为实验眼.每组随机选取3只大鼠视网膜用于免疫印迹法检测,4只大鼠视网膜做HE染色节细胞计数及免疫组化染色荧光强度分析,3只大鼠做免疫电子显微镜观察.采用单因素方差分析对不同结扎时间Ngb蛋白定量及RGCs数目进行统计学分析,采用KSD-t检验进行两两比较.结果 视网膜完全结扎后,A、B、c、D组Ngb蛋白表达量分别为1.439±0.014、2.023±0.134、1.416±0.030及1.073±0.064;RGCs计数结果分别为4368±124、4296 ±96、4132±104、3973±115,组间比较差异有统计学意义（F=79.548,10.191,P〈0.05）.B组表达最高,之后逐渐降低,C组接近正常,此时与A组比较RGCs数量开始减少,D组Ngb的表达已明显低于A组,与A组比较RGCs数量进一步减少.在视网膜各层中,Ngb的表达以RGCs层为最高,其次为内丛状层与外丛状层.RGCs中Ngb主要分布在胞质,C组线粒体外室和线粒体嵴也发现有Ngb表达,而内核层及外核层细胞的胞质内始终未见Ngb的表达.结论 Ngb在大鼠RGCs急性缺血死亡时表达快速升高,主要表达于RGCs的胞质内,与RGCs的生存状态关系密切.Objective To investigate the intra-retinal expression of neuroglobin （Ngb） and death of retinal ganglion cells （RGCs） in acute retina ischemia rats. Methods It was an experimental study. The acute retina ischemia model was established by specific hypodesised left retina artery of Sprague-Dawdey rats. Forty rats were divided into four groups （0, 15, 30, 60 min） by the time of retina ischemia. Every group has 10 rats, in one group random 3 rats were detected by Western blotting;4 rats were detected by ganglion cell counted by hematoxylin and eosin stain and immunohistochemisty fluorescence intensity analysis. The rest 3 rats were detected by Western blotting. The difference among different data were analyzed statistically by One-factor analysis of variance and LSD-t analysis. Results The intra-retinal expression of Ngb reached maximum after acute ischemia 15 minute （P = 0. 000）. then the expression began decreasing. After 30 minute acute ischemia, the expression of Ngb had approached normal （P =0.728）, while, the cell number of RGCs began lower than 0 min group （P = 0.011） ; after 60 minute acute ischemia, the expression of Ngb had been obviously lower than 0 min group （P =0. 001）, the cell number of RGCs had been further lower than 0 min group （P =0. 000）. The expression of Ngb in RGCs layer was highest in rat retina. The expression in inner plexiform layer and external plexiform layer were lower than the former. The expression of Ngb RGCs was mostly intracytoplasm. After 30 minute acute ischemia, the expression of Ngb were detected in mitochondrial outer compartment and mitochondrial cristae, but in cytoplasm of inner nuclear layer and outer nuclear layer the Ngb was not found. Conclusion Ngb quickly steps-up when RGCs die in acute retina ischemia, and mainly expresses intracytoplasm of RGCs. It has tense relationships with nerve cells＇ survival in hypoxia.

关 键 词：视网膜 缺血 神经组织蛋白类 珠蛋白类 视网膜神经节细胞 细胞死亡 免疫组织化学 显微镜检查 免疫电子 

分 类 号：R774.1[医药卫生—眼科]