BMSCs与曲安奈德治疗兔实验性自身免疫性葡萄膜炎的疗效比较  

Comparison of therapeutic effect between bone marrow mesenchymal stem cells and triamcinolone acetonide via intravitreal injection for experimental autoimmune uveoretinitis

作　　者：徐如霞 李静[1] 葛向红[1] 石红霞[1] 孟宪娴[1] 李海燕[1] 杨鹏霞[1] 贾亚丁[1] 

机构地区：[1]山西医科大学&山西省眼科医院,太原030001

出　　处：《中华实验眼科杂志》2014年第7期600-606,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：山西省自然科学基金项目（2011011043-1）

摘　　要：背景 葡萄膜炎多由自身免疫反应所致,以往多应用糖皮质激素和免疫抑制剂进行治疗,但存在药物依赖和眼压升高的风险及其他不良反应.骨髓间充质干细胞（BMSCs）在抗炎、抑制新生血管方面具有较好的作用,且具有低免疫原性和免疫调节功能,但其在葡萄膜炎治疗方面的研究较少. 目的 比较BMSCs和曲安奈德（TA）对兔实验性自身免疫性葡萄膜炎（EAU）的治疗效果.方法 从5只新西兰大白兔股骨和胫骨骨髓中分离、培养BMSCs并进行传代,用流式细胞技术检测细胞表面抗原CD105、CD34和CD45的表达以鉴定细胞,取第3～4代细胞用于实验.将32只新西兰大白兔采用随机数字表法随机分为正常对照组、模型对照组、BMSCs干预组和TA干预组.正常对照组不进行任何处理,其余3个组用质量分数2％牛血清蛋白（BSA）静脉注射和玻璃体腔内注射法建立EAU模型.造模成功后24 h,模型对照组经右眼玻璃体腔内注射0.1 ml PBS; BMSCs干预组以同样方法注射0.1 ml BMSCs悬液,细胞密度为2×106个/ml;TA干预组注射0.1 ml（4 mg）TA.于眼部注射后15d每隔3天裂隙灯显微镜下观察实验兔的眼前节表现,并参照Caspi的标准进行炎症评分;利用眼底彩色照相法及眼部B型超声检查法观察各组兔眼后节表现;于眼部注射后30 d处死实验兔,制备眼组织标本,进行常规组织病理学检查,参照Caspi的标准进行视网膜病理学评分;于眼部注射后每隔3天取各组兔的房水0.1ml及外周血3 ml,采用ELISA法检测兔血清及房水中肿瘤坏死因子-α（TNF-α）的质量浓度.结果 培养的细胞生长状态良好,呈梭形,CD105＋细胞占96.8％,CD34^＋和CD45^＋细胞分别占2.1％和3.5％.正常对照组眼前节、眼后节检查及视网膜组织病理学检查未发现异常.与模型对照组比较,BMSCs干预组和TA干预组兔眼低炎症评分的眼数明显增加,差异均有统计学意义（χ^2=7.25,P=0.01;χ^2=7.37Background Uveitis is caused primarily by autoimmune response,and the conventional therapy is the systemic and topical application of glucocorticoid and immunosuppressive agent.However,these therapies exist a variety of complications.Bone marrow mesenchymal stem cells （BMSCs）are verified to have the anti-inflammation and anti-angiogenesis effects with low immunogenicity and immunoregulation function.But little study is reported for its efficacy in treating uveitis.Objective This study was to compare the therapeutic effects of BMSCs and triamcinolone acetonide （TA）on experimental autoimmune uveoretinitis （EAU）.Methods BMSCs were isolated and cultured from five New Zealand white rabbits,and the third and fourth generation of cells were used in the experiment.The cells were identified by assay of CD105,CD34 and CD45 using flow cytometry.Thirty-two New Zealand white rabbits were randomized into normal control group,model control group,BMSCs intervention group and TA intervention group according to randomized digital table.EAU models were established by intravenous and intravitreal injection of bovine serum albumin （BSA）.After modeling,0.1 ml PBS was intravitreally injected in the model control group,and 0.1 ml BMSCs suspension （2×106/ml）or 0.1 ml （4 mg）TA was used in the same way in the BMSCs intervention group or TA intervention group,respectively.The response of rabbit ocular anterior segment was examined under the slit lamp microscope and scored based on the criteria of Caspi,and the manifestation of ocular posterior segment was evaluated by color fundus photography and B-sonography.The histopathological examination was performed and scored based on the criteria of Caspi 30 days after modeling.Peripheral blood and aqueous humor were obtained for the assay of tumor necrosis factor-α （TNF-α） using ELISA.Results Cultured cells grew well and presented the spindle-like in shape,with the CD105＋cells for 96.8%,CD34^＋cells for 2.1% and CD45^＋cells for 3.5%.No obvious abnormality was se

关 键 词：自身免疫性疾病 动物模型 化学诱导葡萄膜炎 化学诱导视网膜炎 骨髓间充质干细胞 治疗 曲安奈德 治疗 肿瘤坏死因子-α 玻璃体腔内注射 

分 类 号：R773.9[医药卫生—眼科]