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目的探讨神经干细胞(NSCs)移植对视神经受损SD大鼠视网膜节细胞(RGCs)的保护作用。方法将48只健康成年SD大鼠随机分为N组(NSCs移植组)和C组(对照组),均使用精确校准方法在右眼造成部分视神经损伤,左眼作为正常对照。从胚胎SD大鼠海马分离NSCs,利用细胞培养和体内移植技术,将培养后的NSCs注入N组大鼠右眼玻璃体内,C组大鼠右眼玻璃体内注入同等体积的PBS,处死前3d在双上丘注射3%快蓝逆行标记双眼RGCs。将大鼠分别于注射NSCs或PBS后7d、14d、21d、28d处死,各分为4组,每组6只,分离视网膜置于荧光显微镜下,摄影输入计算机图像分析仪计数RGC,计算RGC标识率。另取6只健康成年SD大鼠作NSCs移植,分别于移植后7d、28d处死,每时间段3只,通过视网膜免疫荧光切片观察NSCs在视网膜的存活、整合情况。结果N组大鼠各时间段RGCs标识率与C组同时间段比较均增高,有显著性差异(P<0.01);N组与C组RGCs标识率均随时间延长而降低,且前后段时间比较有显著性差异,但N组降低速度明显慢于C组。NSCs移植7d后部分移植细胞迁移进入视网膜的内网层和节细胞层,28d后可见移植细胞广泛整合至宿主视网膜内。结论NSCs移植入视神经损伤大鼠视网膜后可提高视网膜神经节细胞的存活率,对受损的节细胞具有一定的保护作用。
Objective To investigate the protective effect of neural stem cells (NSCs) transplantation on retinal ganglion cells (RGCs) injured by optic nerve in rats. Methods Forty-eight healthy adult SD rats were randomly divided into N groups (NSCs transplantation group) and C group (control group), all of which were caused by partial retinal damage in the right eye using an accurate calibration method. The left eye was used as a normal control. NSCs were isolated from the hippocampus of embryonic SD rats. The cultured NSCs were injected into the right eye vitreous of N group rats by cell culture and in vivo transplantation. The right eye of C rats were injected with the same volume of PBS into the right eye, Double upper extremity injection of 3% fast blue retrograde labeling of bilateral RGCs. The rats were sacrificed at 7d, 14d, 21d and 28d after injection of NSCs or PBS, respectively. Each group was divided into 4 groups with 6 rats in each group. The retinas were separated and placed under a fluorescence microscope. The RGCs were counted by computerized image analyzer rate. Another 6 healthy adult SD rats were transplanted into NSCs. The mice were sacrificed at 7d and 28d after transplantation respectively. Three rabbits per time period were observed. The survival and integration of NSCs in the retina were observed by immunofluorescence. Results The labeling rate of RGCs in N group was significantly higher than that in C group at the same time (P <0.01). The labeling rate of RGCs in N group and C group decreased with time, There was a significant difference, but the N group was significantly slower than the C group. After 7d transplantation, some of the transplanted cells migrated into the inner reticular layer and the ganglion cell layer of the retina. After 28 days, the transplanted cells were widely integrated into the host retina. Conclusion Transplantation of NSCs into the retina of rats with optic nerve injury can increase the survival rate of retinal ganglion cells and protect the injured ganglion cells.