摘要
目的观察玻璃体内注射色素上皮衍生因子基因修饰的人脐带间充质干细胞(pigment epithelial-derived factor gene-modified human umbilical cord mesenchymal stem cells,PEDF-MSCs)对糖尿病大鼠视网膜组织病理结构改变的影响。方法组织块培养法获取人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,h UCMSCs)。获取的h UCMSCs表达CD105、CD73和CD90,不表达CD34、CD45、CD11b、CD19和HLA-DR。用慢病毒载体以感染复数值为50对其转染。利用链脲佐菌素腹腔注射途径诱导糖尿病大鼠模型,实验动物分为四组:正常对照组(N组)、实验对照组糖尿病注射PBS组(D1组)、糖尿病注射h UCMSCs治疗组(D2组)、糖尿病注射PEDF-MSCs治疗组(D3组)。造模成功后3个月进行干预治疗,N组不予特殊治疗。玻璃体内注射2周后利用荧光显微镜观察PEDF-MSCs在大鼠眼内表达情况。干预治疗2周后进行HE染色观察各层视网膜结构,及各组视网膜总厚度变化。结果玻璃体内注射后2周,荧光显微镜下观察到D2组大鼠玻璃体内簇状排列的红色荧光,视网膜中未发现明显红色荧光;D3组大鼠玻璃体内簇状排列的绿色荧光,视网膜中未发现明显绿色荧光。HE染色显示,N组大鼠的视网膜各层结构完整,层次清晰,细胞排列整齐,染色均匀;D1组大鼠视网膜神经纤维层(nerve fiber layer,NFL)出现明显水肿、血管扩张,内丛状层(innerplexiform layer,IPL)结构疏松,内核层(inner nuclear layer,INL)细胞排列紊乱;D2组NFL水肿减轻;D3组NFL水肿明显减轻,INL与外核层(outer nuclear layer,ONL)层细胞排列规整。视网膜总厚度N组为(103.82±4.15)μm、D1组为(138.86±4.71)μm、D2组为(131.17±3.89)μm、D3组为(112.24±4.22)μm;各组间差异均有统计学意义(均为P<0.05)。结论 PEDF-MSCs可较长时间在糖尿病大鼠玻璃体内存活且持续表达。玻璃体内注射PEDF-MSCs能明显改善糖尿病大鼠视网膜损伤,减轻视网膜水肿。
Objective To investigate the protective effects of intravitreal injection of pigment epithelial-derived factor(PEDF)gene-modified human umbilical cord mesenchymal stem cells(PEDF-MSCs)on the pathological changes in retinal tissue of diabetic rats.Methods hUCMSCs were isolated from human umbilical cord tissue using tissue culture methods,and transfected with lentiviral vectors at a infection multiplicity of 50.Then diabetic model in rats was successfully induced by intraperitoneal injection of streptozotocin.And the rats were divided into normal control(N),PBS treatment(D1),hUCMSCs treatment(D2)and PEDF-MSC treatment(D3)group according to different treatment methods.Three months after modeling,treatment began in D1,D2 and D3 group,but N group left untreated.Two weeks after treatment,the expression of PEDF-MSCs in the eye of rats was detected by fluorescence microscopy,and HE staining was performed to observe the changes in retinal structure and the full-thickness of the retina in each group.Results The expression of CD105,CD73,CD90 was observed,while the expression of CD34,CD45,CD11b,CD19 and HLA-DR did not present.After 2 weeks of treatment,it was in the vitreous cavity not the retina that clusters of red fluorescence appeared in D2 group with fluorescence microscope.There were clusters of green fluorescence in the vitreous cavity not in the retina of D3 group.HE staining showed that the retina had intact structure and clear layers as well as neatly arranged and stained evenly cells in N group.In D1 group,the nerve fibers layer(NFL)showed obvious edema,the blood vessels were dilated,the inner plexiform layer(IPL)were loose and the inner nuclear layer(INL)cells were disordered.In D2 group,the edema of NFL relieved.In D3 group,NFL edema was significantly alleviated,and the cells of INL and outer nuclear layer(ONL)arranged in regular.Full-thickness of retina was(103.82±4.15)μm in N group,(138.86±4.71)μm in D1 group,(131.17±3.89)μm in D2 group,and(112.24±4.22)μm in D3 group,respectively,and the differences were statistically significant(all P<0.05).Conclusion PEDF-MSCs can survive and continue to express in the vitreous cavity of diabetic rats for a long time.Meanwhile,intravitreal injection of PEDF-MSCs can ameliorate retinal edema and the retinal injury in diabetic rats.
作者
韩新红
李艳
李聪伶
单田慧
王淑娜
HAN Xin-Hong;LI Yan;LI Cong-Ling;SHAN Tian-Hui;WANG Shu-Na(Department of Ophthalmology,Weifang Medical College,Weifang 261031,Shandong Province,China;Department of Ophthalmology,Affiliated Hospital of Weifang Medical College,Weifang 261031,Shandong Province,China)
出处
《眼科新进展》
CAS
北大核心
2018年第2期126-130,共5页
Recent Advances in Ophthalmology
基金
山东省高等学校科技计划项目(编号:J15LL08)
潍坊市卫生局科研项目(编号:2014-080)
