[1]韩新红,李艳,李聪伶,等.色素上皮衍生因子基因修饰的脐带间充质干细胞对糖尿病大鼠视网膜组织病理改变的影响[J].眼科新进展,2018,38(2):126-130.[doi:10.13389/j.cnki.rao.2018.0027]
　HAN Xin-Hong,LI Yan,LI Cong-Ling,et al.Pathological changes in retinal tissue in diabetic rats treated with PEDF gene-modified umbilical cord mesenchymal stem cells[J].Recent Advances in Ophthalmology,2018,38(2):126-130.[doi:10.13389/j.cnki.rao.2018.0027]
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色素上皮衍生因子基因修饰的脐带间充质干细胞对糖尿病大鼠视网膜组织病理改变的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
38卷
期数:
2018年2期
页码:
126-130
栏目:
实验研究
出版日期:
2018-02-05

文章信息/Info

Title:
Pathological changes in retinal tissue in diabetic rats treated with PEDF gene-modified umbilical cord mesenchymal stem cells
作者:
韩新红; 李艳; 李聪伶; 单田慧; 王淑娜
261031　山东省潍坊市，潍坊医学院临床学院眼科（韩新红，李艳，李聪伶，单田慧）；261031　山东省潍坊市，潍坊医学院附属医院眼科（李艳，李聪伶，王淑娜）
Author(s):
HAN Xin-Hong; LI Yan; LI Cong-Ling; SHAN Tian-Hui; WANG Shu-Na
Department of Ophthalmology，Weifang Medical College（HAN Xin-Hong，LI Yan，LI Cong-Ling，SHAN Tian-Hui），Weifang 261031，Shandong Province，China；Department of Ophthalmology，Affiliated Hospital of Weifang Medical College（LI Yan，LI Cong-Ling，WANG Shu-Na），Weifang 261031，Shandong Province，China
关键词:
色素上皮衍生因子; 间充质干细胞; 糖尿病视网膜病变; 大鼠
Keywords:
pigment epithelial-derived factor; mesenchymal stem cell; diabetic retinopathy; rats
分类号:
R774
DOI:
10.13389/j.cnki.rao.2018.0027
文献标志码:
A
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，hUCMSCs）。获取的hUCMSCs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hUCMSCs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。
Abstract:
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.

参考文献/References:

［1］　SAADDINE JB，HONEYCUTT AA，NARAYAN KM，ZHANG X，KLEIN R，BOYLE JP.Projection of diabetic retinopathy and the major eye diseases among people with diabetes mellitus：United States，2005-2050［J］.Arch Ophthalmol，2008，126（12）：1740-1747.
［2］　YAU JW，ROGERS SL，KAWASAKI R，LAMOUREUX EL，KOWALSKI JW，BEK T.Global prevalence and major risk factors of diabetic retinophathy［J］.Diabetes Care，2012，35（3）：556-564.
［3］　EZQUER F，EZQUER M，ARANGO-RODRIGUEZ M，CONGET P.Could donormultipotent mesenchymal stromal cell sprevent or delay the onset of diabetic retinopathy［J］?Acta Ophthalmol，2014，92（2）：86-95.
［4］　AMADO LC，SALIARIS AP，SCHULERI KH，STJOHN M，XIE JS，CATTANEO S.Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction［J］.Proc Natl Acad Sci，2005，102：11474-11479.
［5］　JIANG X，ZH Y，LIU B，ZHANG X，WU Y，YU X.Human mesenchymal stem cells inhibit differentiation and function of monocyte-derived dendritic cells［J］.Blood，2005，105（10）：4120-4126.
［6］　YANG Z，LI K，YAN X，DONG F，ZHAO C.Amelioration of diabetic retinopathy by engrafted human adipose-derived mesenchymal stem cells in streptozotocin diabetic rats［J］.Graefes Arch Clin Exp Ophthalmol，2010，248（10）：1415-1422.
［7］　LI G，ZHANG XA，WANG H，WANG X，MENG CL，CHAN CY.Comparative proteomic analysis of mesenchymal stem cells derived from human bone marrow，umbilical cord，and placenta：implication in the migration［J］.Proteomics，2009，9：20-30.
［8］　SI YL，ZHOA YL，HAO HJ，FU XB，HAN WD.MSC：biological characteristics，clinical applications and their outstanding concerns［J］.Ageing Res Rev，2011，10：93-103.
［9］　SCALINCI SZ，CALINCI SZ，SCOROLLI L，CORRADET TI.Potential role of intravitreal human placental stem cell implants in inhibiting progression of diabetic retinopathy in type 2 diabetes：neuroprotective growth factors in the vitreous［J］.Clin Ophthalmol，2011，5：691-696.
［10］　YAMAGIS HI，MATSUI T，NAKAMURA K，UEDA S，NODA Y，IMAIZUMI T.Pigment epithelium-derived factor （PEDF）：its potential therapeutic implication in diabetic vascular complications［J］.Curr Drug Targets，2008，9（11）：1025-1029.
［11］　ZHU XF，ZOU HD.PEDF in diabetic retinopathy：a protective effect of oxidative stress［J］.Biomed Biotechnol，2012，（2012）：580687.
［12］　SHEN X，XIE B，CHENG Y，JIAO Q，ZHONG Y.Effect of pigment epithelium derived factor on the expression of glutamine synthetase in early phase of experimental diabetic retinopathy［J］.Ocul Immunol Inflamm，2011，19（4）：246-254.
［13］　SHEN X，ZHONG Y，XIE B，CHENG Y，JIAO Q.Pigment epithelium derived factor as an anti-inflammatory factor against decrease of glutamine synthetase expression in retinal müller cells under high glucose conditions［J］.Graefes Arch Clin Exp Ophthalmol，2010，248（8）：1127-1136.
［14］　XIE B，JIAO Q，CHENG Y，ZHONG Y，SHEN X.Effect of pigment epithelium-derived factor on glutamate uptake in retinal müller cells under high-glucose conditions［J］.Invest Ophthalmol Vis Sci，2012，53（2）：1023-1032.
［15］　LI MY，LI Y，LIU HB，BI HL，ZHANG J.Effects of PEDF on rat retinal Müller cellsapoptosis stimulated by high glucose［J］.Rec Adv Ophthalmol，2014，34（6）：526-529.
李梦云，李艳，刘宏波，毕欢丽，张杰.色素上皮衍生因子对高糖刺激下大鼠视网膜Müller细胞凋亡的影响［J］.眼科新进展，2014，34（6）：526-529.
［16］　PARK HY，KIM JH，SUN KH，PARK CK.Stem cell-based delivery of brain-derived neurotrophic factor gene in the rat retina［J］.Brain Res，2012，1469（1）：10-23.

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备注/Memo

备注/Memo:
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）
更新日期/Last Update: 2018-02-02