间充质干细胞来源外泌体(MSCs-exo)对大鼠角膜移植排斥反应的抑制作用  

Suppressive effect of exosomes derived from mesenchymal stem cells on keratoplasty rejection

作　　者：贾喆 李斐 吕瑛 曾孝宇 路晓晓 赵少贞 JIA Zhe;LI Fei;LV Ying;ZENG Xiao-Yu;LU Xiao-Xiao;ZHAO Shao-Zhen(Tianjin Medical University Eye Hospital,Tianjin 300384,China)

机构地区：[1]天津医科大学眼科医院,天津市300384

出　　处：《眼科新进展》2018年第11期1010-1013,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:81770890;81600705);天津市自然科学基金(编号:16JONJC12500)~~

摘　　要：目的观察间充质干细胞来源外泌体(mesenchymal stems cells cderived exosomes,MSCs-exo)对大鼠角膜移植排斥反应的抑制作用。方法制作大鼠异体角膜移植排斥反应模型,随机分为4组:空白对照组、MSCs结膜下治疗组、PBS结膜下注射组、MSCs-exo结膜下治疗组。术后第3天开始裂隙灯下观察角膜植片排斥情况,并对MSCs-exo进行染色及示踪观察其结局。结果空白对照组角膜植片存活时间为(9.67±0.56)d,MSCs结膜下治疗组为(12. 33±0. 76)d,PBS结膜下注射组为(9. 50±0.34)d,MSCs-exo结膜下治疗组为(16. 5±1.15) d。PBS结膜下注射组角膜植片存活时间较空白对照组差异无统计学意义(P>0.05),而MSCs结膜下治疗组与MSCs-exo结膜下治疗组角膜植片存活时间较PBS结膜下注射组均明显延长(均为P <0. 05),MSCs-exo结膜下治疗组植片存活时间长于MSCs结膜下治疗组(P<0. 05)。空白对照组大鼠免疫组织化学染色结果显示植片部分厚度比植床角膜厚度显著增厚,上皮层及基质层内见大量CD4+细胞聚集。而MSCs-exo结膜下治疗组植片中CD4+细胞比空白对照组明显减少。MSCs-exo结膜下注射后72 h时,仅有少量外泌体可在结膜下被观测到。进一步将观测时间前推至注射后24 h,发现大量外泌体存在于结膜下,而角膜组织和前房内也可见到外泌体的定位表达。结论结膜下注射MSCs-exo可以显著延长角膜植片存活时间,外泌体治疗可能成为无细胞治疗的新方法。Objective To investigate the effect of mesenchymal stem cells derived exosomes(MSC-exo)on keratoplasty rejection in a rat model.Methods A Wistar-Lewis rat corneal transplant rejection model(Wistar rats to Lewis rats)was established and then randomly separated into four groups:negative control group,MSCs treatment group,PBS control group and MSC-exo treatment group.Graft rejection was scored using slit lamp since the 3rd day after the operation.Labeled MSC-exo was used for tracking during the experiments to demonstrate the outcome of the exosomes.Results The survival time of the corneal grafts were(9.67±0.56)days for negative control group,(12.33±0.76)days for MSCs treatment group,(9.50±0.34)days for PBS control group,and(16.5±1.15)days for MSC-exo treatment group.Postoperative MSCs and MSCs-exo treatment groups prolonged graft survival time when compared with PBS control group,and the differences were statistically significant(all P<0.05).And the suppressive effect of MSCs-exo was stronger than MSCs(P<0.05).MSC-exo treatment groups also showed suppression of CD4+T cell infiltration in the allograft region in immunohistochemistry tests.Only a small number of exosomes could be observed 72 hours after the subconjunctival injection.However,a large number of labeled MSC-exo could be found when observation time was pushed forward to 24 hours after injection.Moreover,the labeled exosomes could also be found in corneal tissue and anterior chamber.Conclusion Subconjuctival injection of MSC-exo ccan prolong the survival time of the corneal allograft which indicates MSCs-exo can be used as a novel treatment approach in cell-free therapy.

关 键 词：间充质干细胞 外泌体 角膜移植排斥 无细胞治疗 

分 类 号：R772.2[医药卫生—眼科]