【作者】 苗刚勇；

【导师】 唐罗生；

【作者基本信息】 中南大学 ， 眼科学， 2008， 硕士

【摘要】 目的:1.探讨MCP-1在Brown Norway大鼠试验性视网膜脱离中的表达,及其与视网膜脱离过程中光感受器细胞凋亡的相关性。2.探讨干预脱离的视网膜MCP-1的表达,光感受器细胞凋亡的变化。方法:(1)取37只Brown Norway大鼠右眼建立视网膜脱离模型,左眼自身对照。排除因死亡,眼内炎和视网膜出血等不符合视网膜脱离模型标准的大鼠7只。剩余30只大鼠按预处理时间点(1小时,6小时,12小时,24小时和72小时)随机分为五组,每组6只。预定时间点处死大鼠,取出眼球。免疫组化技术检测实验眼与对照眼视网膜外核层MCP-1的表达;TUNEL原位末端标记法检测实验眼与对照眼外核层凋亡细胞阳性表达细胞数。统计软件分析MCP-1与TUNEL表达的相关性。(2)9只Brown Norway大鼠双眼建立视网膜脱离模型后,沿原穿刺口右眼视网膜下注射抗大鼠MCP-1 IgG抗体,左眼注射等容积的PBS。排除因死亡,眼内炎和视网膜出血等不符合视网膜脱离模型标准的大鼠3只。剩余6只大鼠模型建立后72小时处死,剜出眼球,制作石蜡切片,应用TUNEL原位末端标记法计数外核层阳性细胞。统计学方法比较72小时视网膜脱离组,MCP-1干预组和假干预组的外核层凋亡细胞的表达。结果:(1)免疫组化:正常视网膜MCP-1很少表达,点状分布于内核层,外核层和神经节细胞层,每个高倍镜下外核层阳性表达细胞数平均为3.64±1.02。试验性视网膜脱离模型建立1小时、6小时、12小时、24小时和72小时后MCP-1表达主要分布于外核层,内核层与神经节细胞层较少表达,外核层高倍镜下MCP-1阳性表达细胞数分别为10.50±2.82、11.13±3.31、25.23±5.56、37.23±7.15和205.83±15.23,与对照组均有统计学差异。视网膜脱离组之间相互比较,除视网膜脱离1小时和6小时无统计学差异外,其余各组均有显著性差异(P＜0.05)。(2)TUNEL:正常视网膜外核层TUNEL阳性细胞数为2.16±1.33/mm~2,试验性视网膜脱离模型建立1小时、6小时、12小时、24小时和72小时外核层TUNEL阳性表达细胞数分别为15.30±3.21/mm~2、25.11±5.31/mm~2、41.32±7.56/mm~2、80.23±10.15/mm~2和425.83±25.23/mm~2。(3)各组大鼠实验眼视网膜ONL内MCP-1及TUNEL阳性细胞均于RD后1h开始升高,随时间延长表达增加,72小时达到高峰,二者的表达呈显著正相关(r=0.905)。(4)视网膜脱离后72小时MCP-1干预组和PB S假干预组外核层TUNEL阳性表达细胞数为160.24±10.56/mm~2和430.15±30.42/mm~2。MCP-1干预组与视网膜脱离组统计学上具有显著性差异(p＜0.05),而PBS假干预组与视网膜脱离组统计学无显著性差异(p＞0.05)。结论:(1)MCP-1在大鼠试验性视网膜脱离模型建立后1小时即明显表达,随时间延长表达增加,72小时达到高峰。(2)视网膜脱离各时相的MCP-1与TUNEL阳性表达细胞数具有明显正相关。(3)干预脱离的视网膜MCP-1的表达可减少光感受器细胞的凋亡。更多还原

【Abstract】 Objective:(1)To explore Monocyte chemoattractant protein-1 expresses in rat experimental retinal detachment at different times;the relationship between MCP-1 and photoreceptor apoptosis in retinal detachment.(2)To explore blocking the expression of MCP-1 in retinal detachment can diminish photoreceptor apoptosis.Methods:46 Brown Norway rats were involved in this study.(1)37 rats were builded the retinal detachment model in the right eye,and the left eye as control.Except 7 rats which were not fit to the standard model(include death, endophthalmitis and retinal hemorrhage).The left 30 rats devided into 5 groups according force-managed times(1h、6h、12h、24h、72h).Then rats were killed and the eyes were enclutered.The expression of MCP-1 in retinal detachment eyes were assessed by immunohistochemical method and cell death were quantified by TUNEL assay.Analysis the relatinship between MCP-1 and TUNEL in detached retinal by statistical software.(2)9 rats were builded the retinal detachment model both of the eyes.We injected rat anti-MCP-1 IgG subretinally during RD induction in the right eyes and PBS in the left eyes. Except 3 rats which were not fit to the standard model(include death,endophthalmitis and retinal hemorrhage).Cell death were quantified by TUNEL assay 72h after RD,since at that time the number of TUNEL-positive cells in the ONL is at the peak.Resuits(1)Immunohistochemical analysis:In normal retinal tissue,the number of MCP-1 positive cells was 3.64±1.02 under microscopy visual field.MCP-1 positive cells was 10.50±2.82、11.13±3.31、25.23±5.56、37.23±7.15、205.83±15.23 in detached retinal at 1h、6h、12h、24h、72h under microscopy visual field.Compare the expression of MCP-1 at different times of RD,every two groups had statistical variation,except the 1h and 6h group.(2)TUNEL assey:In normal retinal tissue,the number of TUNEL positive cells in ONL was 2.16±1.33/mm~2.TUNEL positive cells in ONL was 15.30±3.21/mm~2;25.11±5.31/mm~2;41.32±7.56/mm~2; 80.23±10.15/mm~2;425.83±25.23/mm~2 in detached retinal at 1h、6h、12h、24h、72h.(3)MCP-1 and TUNEL positive cell began to express remarkably after 1h of retinal detachment,then increased follow time,got the peak at 72h.The relationship between MCP-1 and TUNEL positive cell is positive correlation(r=0.905).(4)TUNEL positive cells in MCP-1 blocking group and PBS group after 72h of retinal detachment were 160.24±10.56/mm~2 and 430.15±30.42/mm~2 in ONL.There was statistical variation between MCP-1 blocking group and RD group(p＜0.05),no statistical variance between RD group and PBS group(p＞0.05).Conclution:(1)MCP-1 began to express after 1h of rat retinal detachment,then increased follow time,got the peak at 72h.(2)There is positive correlation between MCP-1 and TUNEL positive cell.(3)Blocking the expression of MCP-1 in detached retinal can diminish photoreceptor apoptosis.更多还原