摘要
目的观察髓磷脂抑制蛋白Nogo-A/B在遗传性视网膜色素变性(retinitis pigmentosa,RP)遗传模型皇家外科学院大鼠(Royal college of surgeons rat,RCS)视网膜变性发展过程中的表达并探讨其意义。方法按出生后视网膜变性的发展程度将RCS-p+(视网膜含色素的变性大鼠)分为P15d、P30d、P60d和P90d 4组,采用相应时期的RCS-rdy+p+(视网膜含色素的正常大鼠)作为对照,5只/时间点,利用免疫组织化学技术及免疫印迹检测2组(共40只)RCS视网膜上Nogo-A/B蛋白的表达趋势。结果与对照组相比,①P15d、P30d RCS-p+大鼠视网膜各层结构和细胞数目无明显变化,仅节细胞数目轻微减少;P60d、P90d大鼠视网膜外核层(outer nuclear layer,ONL)和内核层(inner nuclear layer,INL)结构出现明显紊乱,细胞数目的减少以ONL和神经节细胞层(ganglion cell layer,RGC)层为主。②IH显示Nogo-A/B蛋白在RCS-p+大鼠各时间段视网膜表达均为阳性,P15d开始出现,随变性过程呈升高趋势,主要表达在INL和RGC层,阳性细胞数目显著高于对照组。③WB免疫印迹检测结果显示P15d、P30d、P60d和P90d Nogo-A蛋白表达量分别为(0.827 37±0.212 92)、(1.110 19±0.089 99)、(1.315 52±0.028 57)、(1.268 81±0.080 42),与对照组相比有显著差异(P<0.05)。Nogo-B2蛋白与Nogo-A蛋白表达趋势一致,但总体表达量较低。结论 Nogo-A蛋白可能参与了视网膜色素变性过程中视神经损伤后的再生修复抑制。
Objective To investigate the expression of Nogo-A/B proteins in the retina of Royal college of surgeons（RCS） rats（a classic genetic animal model with hereditary retinal degeneration） and its significance.Methods According to the development status of retinal degeneration,the RCS-p＋（RCS rats with inherited retinal dystrophy） rats were divided into 4 groups including a P15d group,a P30d group,a P60d group and a P90d group,and RCS-rdy＋p＋（RCS rats without retinal degeneration） rats of the corresponding periods were used as control groups（C15d group,C30d group,C60d group and C90d group,n=5）.Immunohistochemical assay and Western blot analysis were used to detect the expression patterns of Nogo-A/B proteins in the retinal tissues of the RCS rats（RCS-p＋/RCS-rdy＋p＋）.Results（1） Compared with the control groups,no obvious change of the cell numbers and thickness of retina were observed in the P15d and P30d groups,except for a slight decrease of retinal ganglion cell（RGC） number.In the P60d and P90d groups,apparent structural disorder was observed in the outer nuclear layer（ONL） and inner nuclear layer（INL） of the retina.In addition,the cell number decrease was significant in the ONL and RGC layers.（2） The immunohistochemistry results showed that the positive expression of Nogo-A/B proteins was detected in each time period,and the Nogo-A/B expression upregulated along with the retinal degeneration process from P15d,mainly locating in the INL and RGC layers.The number of Nogo-A/B-positive cells was significantly greater in the RCS-p＋ rats than in the RCS-rdy＋p＋ rats.（3） Western blot analysis showed that the protein expression of Nogo-A of the RCS-p＋ rats at P15d,P30d,P60d and P90d were（0.827 37±0.212 92）,（1.110 19±0.089 99）,（1.315 52±0.028 57） and（1.268 81±0.080 42）,respectively,which were significantly different from those of the control groups（P0.05）.The trend of Nogo-B2 protein expression was consistent with that of the Nogo-A prote
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2012年第10期948-952,共5页
Journal of Third Military Medical University
