嗅鞘细胞与睫状神经营养因子协同促进视神经损伤大鼠视神经修复的研究  

Synergetic effects of ciliary neurotrophic factor and olfactory ensheathing cells on optic nerve reparation

作　　者：尹丹萍[1] 柳林[2] 曹莉[3] 

机构地区：[1]济南军区总医院眼科,250031 [2]上海交通大学医学院附属仁济医院眼科 [3]第二军医大学神经生物教研室

出　　处：《中华眼科杂志》2013年第11期1020-1028,共9页Chinese Journal of Ophthalmology

摘　　要：目的 研究大鼠视神经吸断后嗅鞘细胞（OEC）和睫状神经营养因子（CNTF）单独及联合应用对视网膜神经节细胞（RGC）及其轴突的保护及促进修复作用.方法 实验研究.采用极细微玻管吸断成年大鼠视神经制作视神经吸断伤动物模型,将实验大鼠分为4组：对照组、OEC组、CNTF组和OEC ＋CNTF联合应用组.对照组为未行治疗的视神经损伤组,OEC组为视神经吸断伤处注入OEC组,CNTF组为视神经吸断伤后玻璃体内注射CNTF组,OEC＋ CNTF联合应用组是视神经吸断伤处注入OEC和玻璃体内注射CNTF共同应用组.术后4周,每组中大鼠全部玻璃体内注射霍乱毒素B亚单位（CTB）,顺行示踪标记视神经轴突,并联合在视神经远侧端注入荧光金或生物素化葡聚糖胺（BDA）逆行示踪标记视网膜神经节细胞,计算各组实验大鼠相同视野下视网膜神经节细胞即神经元的存活数、神经元轴突密度.对数据进行方差分析.结果 各治疗组视网膜神经节细胞存活数明显多于对照组,OEC组的视网膜神经节细胞存活数高于CNTF组,OEC和CNTF联合应用组视网膜神经节细胞存活数最高,各组RGC平均数分别为：OEC ＋CNTF组46.00±1.42,OEC组34.80±1.34,CNTF组25.00±1.61,对照组15.40±2.30,差异有统计学意义（F=633.38,P＜0.01）.各治疗组神经元轴突密度也大于对照组,OEC和CNTF联合应用组神经元轴突密度最大.各组视神经轴突平均光密度（A值×1O-2）分别为OEC＋ CNTF组3.18 ±0.26、OEC组2.96±0.28、CNTF组2.83±0.37、对照组2.75 ±0.12,差异有统计学意义（F=17.66,P ＜0.01）.结论 局部应用外源性OEC和玻璃体注射CNTF均能减轻视神经损伤引起的逆行性神经元损害,对视神经元有保护作用.OEC保护视神经元的作用优于CNTF.OEC与CNTF联合应用保护视神经元效果更加显著,两者合用产生了协同作用.Objective This study is to investigate the effect of the combination of the olfactory ensheathing cells （OEC） transplantation and intravitreous injection of ciliary neurotrophic factor （CNTF） on the retinal ganglial cells＇ （RGC） apoptosis and axonals＇ reparation and regeneration.Methods In this study,the supraorbital margin exposure of the optic nerves was used to establish adult SD rats＇ optic nerve inhausted injury model as control group.Then the purified OECs were injected into the optic nerve sheaths,and CNTF was injected into the vitreous body simultaneously.The rats were divided into control group,CNTF group,OECs group,and OEC ＋ CNTF combined group.At 4 weeks postoperatively,a cholera toxin B subunit （CTB） anterograde tracing technique and fluorescence （FG） biotinylated dextran amine （BDA） retrograde tracing technique were adopted to evaluate the survival of RGC and the regeneration of optic nerve axons.The number of survival neurons of the same vision field and the density of neurons were evaluated by analysis of variance.Results At the one and three quadrant distance between optic disc 2 mm spot,the number of the RGC in the control group was significantly （F =633.38,P ＜ 0.01） decreased compared with the normal group（46.00 ± 1.42,34.80 ± 1.34,25.00 ± 1.61,15.40 ± 2.30）.The survived RGC in OEC group was significantly more than that in CNTF group,while the combined treatment with CNTF and OEC had strongest repair effect.The neuron axon density showed a statistically significant difference in the average optical density value at distance between foramen opticum 2 mm spot （OEC ＋ CNTF：3.18 ± 0.26,OEC：2.96 ± 0.28,CNTF：2.83 ± 0.37,and control：2.75 ± 0.12,respectively,F =17.66,P ＜ 0.01）.Conclusions Optic nerves damage can be repaired and regenerated by the combined treatment with OEC and CNTF suggesting that CNTF and OEC have synergistic effect on the treatment of optic nerve injury and repair.Transplantation of OECs may genetically modify the secre

关 键 词：视神经损伤 嗅球 细胞移植 睫状神经营养因子 视网膜神经节细胞 细胞存活 

分 类 号：R774.6[医药卫生—眼科]