[1]李娟,周颖,谭钢,等.间充质干细胞外泌体治疗小鼠干眼的疗效评价[J].眼科新进展,2019,39(10):901-905.[doi:10.13389/j.cnki.rao.2019.0205]
 LI Juan,ZHOU Ying,TAN Gang,et al.Therapeutic effects of mesenchymal stem cells derived exosomes on dry eye mice[J].Recent Advances in Ophthalmology,2019,39(10):901-905.[doi:10.13389/j.cnki.rao.2019.0205]
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
39卷
期数:
2019年10期
页码:
901-905
栏目:
实验研究
出版日期:
2019-10-05

文章信息/Info

Title:
Therapeutic effects of mesenchymal stem cells derived exosomes on dry eye mice
作者:
李娟周颖谭钢王甜鲜盼盼龙乾发
710004 陕西省西安市,西安市第四医院眼科,陕西省眼科医院,西安交通大学附属广仁医院(李娟);415003 湖南省常德市,常德市第一人民医院眼科(周颖);421001 湖南省衡阳市,南华大学附属第一医院眼科(谭钢);710001 陕西省西安市,西安市中心医院神经外科(王甜,鲜盼盼,龙乾发)
Author(s):
LI JuanZHOU YingTAN GangWANG TianXIAN Pan-PanLONG Qian-Fa
Department of Ophthalmology,Xi’an No.4 Hospital,Shaanxi Eye Hospital,Affiliated Guangren Hospital School of Medicine,Xi’an Jiaotong University(LI Juan);Department of Ophthalmology,the First People’s Hospital of Changde (ZHOU Ying);Department of Ophthalmology,the First Affiliated Hospital of the University of South China (TAN Gang);Department of Neurosurgery,Xi’an Central Hospital(WANG Tian,XIAN Pan-Pan,LONG Qian-Fa)
关键词:
外泌体干眼泪膜功能
Keywords:
exosomesdry eyetear film function
分类号:
R772.2
DOI:
10.13389/j.cnki.rao.2019.0205
文献标志码:
A
摘要:
目的 探讨人脐带血间充质干细胞外泌体(mesenchymal stem cells derived exosomes,MSC-Exo)治疗小鼠干眼的效果。方法 选取32只(64眼)雄性6~8周龄BALB/c小鼠,随机分为A、B、C、D组,每组各8只。除D组不予处理外,其余24只均给予2 g·L-1苯扎氯铵滴双眼,连续14 d,各建立中重度干眼模型。造模结束后,A组用25 g·L-1MSC-Exo滴眼液点眼,B组用磷酸盐缓冲液点眼,C组为阴性对照组,造模后不予处理。分别在治疗前和治疗后第1、4、7天进行小鼠泪膜功能相关检查,包括泪液分泌实验(SchirmerⅠtest,S Ⅰ T)、泪膜破裂时间(tear break-up time,BUT)、荧光素染色评分(corneal fluorescein staining,FL)测定。于相应时间点收集其角膜组织,行苏木精-伊红染色及透射电子显微镜下观察角膜上皮组织结构变化。结果 治疗前及治疗后第1天,A、B、C三组S Ⅰ T、BUT及FL(FL仅A、B组比较)差异均无统计学意义(均为P>0.05),各时间点S Ⅰ T、BUT A、B、C组与D组差异均有统计学意义(均为P<0.05)。治疗后第4天、第7天,A组S Ⅰ T较B组、C组明显增加,A组BUT较B组、C组明显延长,差异均有统计学意义(均为P<0.05)。治疗后第4天、第7天,A组S Ⅰ T较B组,C组明显增加,A组BUT较B组、C组明显延长,差异均有统计学意义(均为P<0.05)。治疗后第4天和第7天,A组S Ⅰ T、BUT均较治疗前明显增加(均为P<0.05),FL评分较治疗前降低(P<0.05);而B组S Ⅰ T、BUT与治疗前差异均无统计学意义(均为P>0.05),FL差异无统计学意义(P>0.05);C组造模后未行治疗;D组未行任何处理,两组各项检测指标均未见明显变化(均为P>0.05)。电镜观察可见:A组角膜上皮微绒毛呈指状突起,微绒毛数量多,排列整齐;B组角膜上皮微绒毛减少、变短、排列紊乱;C组角膜上皮微绒毛可见明显减少、脱落、缺失,排列紊乱;D组角膜上皮微绒毛形态呈指状突起,排列整齐,数量多;C组与A组和D组有很大差异。结论 MSC-Exo滴眼液对小鼠干眼有明显的治疗作用,有望为临床干眼的治疗带来新方法。
Abstract:
Objective To investigate the therapeutic effects of human umbilical cord mesenchymal stem cells derived exosomes (MSC-Exo) on a mouse model of dry eye.Methods Thirty-two healthy adult male BALB/c mice (64 eyes) aged 6-8 weeks were selected in this study and divided into four groups randomly (group A,B,C and D),with each group consisting of 8 mice.The dry eye model was induced by eye drop of 2 g·L-1 benzalkonium chloridefor 14 consecutive days in group A,group B and group C.Group D served as the normal controls without any treatment.The A group was given 25 g·L-1 MSC-Exo eye drops,while the group B was given PBS eye drops and the group C were not received any treatment as the negative control.The Schirmer I test(SIT),tear break-up time (BUT) and corneal fluorescein staining (FL) were evaluated before and after eye drops on day 1,day 4 and day 7.The cornea samples of all mice were collected for histological investigation of corneal epithelium using hematoxylin and eosin staining,and ultrastructure of corneal epithelial cells was further examined by transmission electron microscopy (TEM).Results There were no statistical changes in SIT,BUT before therapy and 1 day after treatment among group A,group B and group C,and no statistical changes in FL between group A and group B at both time points.Moreover there were significant differences in SIT,BUT between group A,B,C and group D (all P<0.05).Significantly more tear volume and longer BUT were shown on day 4 and day 7 after treatment in group A than group B and group C(all P<0.05).On day 4 and day 7 after treatment,SIT and BUT were significantly increased in group A (both P<0.05),and FL score was lower than before treatment (P<0.05).There were no significant differences in SIT,BUT,FL before and after treatment in group B (all P>0.05).No treatment was performed after modeling in group C,and no treatment was performed in group D.There were no significant changes in parameters of the two groups (all P>0.05).TEM showed that the number of microvilli of corneal epithelium was large in group A,and they arranged neatly and had finger-like protrusions;the microvilli in group B decreased,shortened and arranged disorderly;they were significantly reduced,shedding,and missing,and their arrangement was disordered in group C,while the microvilli in group D were finger-like,neatly arranged and numerous.Group C was very different from group A and group D.Conclusion The exosomes derived from mesenchymal stem cells exhibit signicantly therapeutic effects on dry eye mice model,which suggest that MSC-Exo administration is a promising method for the treatment of dry eye.

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备注/Memo

备注/Memo:
国家自然科学基金资助(编号:81400424、81871084);2017年陕西省创新人才推进计划项目-青年科技新星项目(编号:2017KJXX-87);陕西省科学技术研究发展计划项目(编号:2014K11-03-07-04);中央高校基本科研业务费专项资金资助(编号:xzy012019122)
更新日期/Last Update: 2019-10-12