摘要
目的研究骨髓间充质干细胞移植对视神经损伤大鼠视网膜细胞凋亡的作用。方法制作大鼠视神经钳夹伤模型,随机分为干细胞移植治疗组(M组)和磷酸缓冲液治疗对照组(P组),取大鼠股骨和胫骨骨髓,密度梯度离心结合贴壁筛选法分离纯化大鼠骨髓间充质干细胞(MSCs)并鉴定,将传代培养后的定量MSCs注入损伤术后M组大鼠双眼玻璃体内,P组大鼠则注入等体积PBS,观察视神经损伤后1 d3、d7、d、14 d2、1 d和28 d视网膜形态学改变并应用流式细胞仪检测视网膜细胞凋亡率变化。结果视神经损伤后视网膜神经节细胞(retinal ganglion cell,RGCs)数目随时间延长而减少,组内前后时间段比较有显著性差异(P<0.05);M组RGCs降低速度明显慢于P组,M组各时间段RGCs数目与P组同时间段相比均增高,有显著性差异(P<0.05);视神经损伤后视网膜细胞出现凋亡,1 d-14 d内随时间延长凋亡率不断上升,14 d达到高峰,随后下降;损伤后21 d内M组各时间段细胞凋亡率均低于P组,二者比较有显著性差异(P<0.05),损伤后28 d二者的细胞凋亡率没有统计学差异(P>0.05)。结论 MSCs玻璃体内移植可提高视神经损伤大鼠RGCs的存活率,抵抗视网膜细胞凋亡。
Objective To investigate the effect of mesenchymal stem cell（MSCs） transplantation on retinal ganglion cells（RGCs）of Sprague-Dawley（SD）rats with optic nerve injury.Methods Optic nerve crush injury models were induced in the eyes of sixty-six adult SD rats by a calibrated method,which were randomly divided into group M（MSCs） and group P（PBS）.MSCs were isolated from SD rats by gradient centrifugation and anchoring culture method.Immunocytochemical staining was used to explore the surface antigen marker.The morphological changes of optic nerves were observed and the number of RGCs were examined by HE staining at 1 d、3 d、7 d、14 d、21 d and 28 d after optic nerve injury.At the same time,cell apoptosis rate of retinal was measured by flow cytometery.Results The number of RGCs was reduced significantly after optical nerve injury.The speed of RGCs decline in M group was more slowly than that of the P group,the number of RGCs in M group were higher compared with the P group at the same injury period,there was statistical significance between them（P〈0.05）.The rate of cell apoptosis increased and reached peak at 14d after optical nerve injury and then decreased.The apoptotic rate of M group were lower than that of P group at 1 d、3 d、7 d、14 d、21 d,there was statistical significance between them（P〈0.05）.At 28 d after injury,there was no statistical significance between M group and P group（P〈0.05）.Conclusion MSCs transplantation could rescue the survival rate of injured RGCs in optic nerve-crush rat and resist the cell apoptosis of retinal to realize it＇s protective effect.
出处
《中国实验诊断学》
北大核心
2010年第7期987-990,共4页
Chinese Journal of Laboratory Diagnosis
基金
广东省医学基金资助项目(A2008137)
