[1]李　鑫,洪　萌,侯铁奇.骨髓间充质干细胞对视网膜色素变性大鼠视网膜的修复作用[J].新乡医学院学报,2021,38(6):511-515.[doi:10.7683/xxyxyxb.2021.06.003]
　LI Xin,HONG Meng,HOU Tieqi.Repair effect of bone marrow mesenchymal stem cells on the retina of rats with retinitis pigmentosa[J].Journal of Xinxiang Medical University,2021,38(6):511-515.[doi:10.7683/xxyxyxb.2021.06.003]
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骨髓间充质干细胞对视网膜色素变性大鼠视网膜的修复作用
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年6
页码:
511-515
栏目:
基础研究
出版日期:
2021-06-05

文章信息/Info

Title:
Repair effect of bone marrow mesenchymal stem cells on the retina of rats with retinitis pigmentosa
作者:
李　鑫1; 洪　萌2; 侯铁奇3
（1.新乡市中心医院眼科，河南　新乡　453000；2.河南大学第一附属医院眼科，河南　开封　475000；3.中山大学附属第三医院眼科中心，广东　广州　510630）
Author(s):
LI Xin1; HONG Meng2; HOU Tieqi3
(1.Department of Ophthalmology,Xinxiang Central Hospital,Xinxiang 453000,Henan Province,China2.Department of Ophthalmology,the First Affiliated Hospital of Henan University,Kaifeng 475000,Henan Province,China3.Ophthalmic Center of the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,Guangdong Province,China)
关键词:
骨髓间充质干细胞; 视网膜色素变性; 视网膜修复
Keywords:
bone marrow mesenchymal stem cellretinitis pigmentosaretinal repair
分类号:
R774.1+3
DOI:
10.7683/xxyxyxb.2021.06.003
文献标志码:
A
摘要:
目的　探讨大鼠骨髓间充质干细胞（BMSC）的分离制备方法及其对视网膜色素变性（RP）大鼠视网膜的修复作用。方法　选取Sprague-Dawley大鼠2只，采用贴壁法分离获取BMSC，采用生长曲线、流式细胞术及细胞形态观察鉴定BMSC，并制备BMSC悬液。将45只Sprague-Dawley大鼠随机分为对照组、RP组和BMSC组，每组15只。对照组大鼠正常饲养，不制备RP大鼠模型。RP组和BMSC组大鼠采用碘酸钠（NaIO3）法建立RP大鼠模型。建模成功后，RP组大鼠局部视网膜下注射0.15 mL生理盐水，BMSC组大鼠局部视网膜下注射BMSC悬液0.15 mL（细胞密度为1×109 L-1），每日1次，连续注射7 d；对照组大鼠不采取任何干预措施。分别于造模前、造模后及干预后1个月检测3组大鼠血清脑源性神经营养因子(BDNF) 、白细胞介素-10（IL-10）水平；干预后1个月，采用断颈法处死大鼠，取大鼠视网膜组织，苏木精-伊红（HE）染色法观察各组大鼠视网膜病变部位组织形态。结果　倒置显微镜观察显示，制备的BMSC培养2 h后开始贴壁生长，培养2 d后伸出伪足，且长短不一，培养4 d后快速增长，细胞开始集落排列，细胞质丰富；细胞传代后分布相对均匀，形态纤长，细胞排列一致，呈旋涡或放射状。细胞生长曲线结果显示，BMSC接种第1～2 天为潜伏期，细胞生长相对缓慢；BMSC接种第3～5天为对数生长期，细胞生长迅速。第3代大鼠BMSC流式细胞术检测显示，CD45+、CD29+、CD44+细胞率分别为1.33%、79.95%、81.49%。造模前3组大鼠血清BDNF、IL-10水平比较差异无统计学意义（P＞0.05），造模后RP组和BMSC组大鼠血清BDNF、IL-10水平显著低于对照组（P＜0.05）。RP组和BMSC组大鼠造模后血清BDNF、IL-10水平显著低于造模前（P＜0.05），BMSC组大鼠干预后血清BDNF、IL-10水平显著高于造模后（P＜0.05）；干预后，BMSC组大鼠血清BDNF、IL-10水平显著高于RP组（P＜0.05）；对照组大鼠造模前、造模后、干预后血清BDNF、IL-10水平比较差异无统计学意义（P＞0.05）。HE染色显示，对照组大鼠视网膜细胞排列整齐，细胞完好；RP组大鼠视网膜细胞排列紊乱，存在许多细胞碎片；与RP组比较，BMSC组大鼠视网膜细胞排列相对整齐，细胞碎片显著减少。结论　BMSC可以显著提高RP大鼠血清BDNF、IL-10水平，促进受损视网膜修复。
Abstract:
Objective　To investigate the isolation and preparation methods of rat bone marrow mesenchymal stem cells (BMSC) and its repair effect on the retina of rats with retinitis pigmentosa (RP).Methods　　BMSCs were isolated from two Sprague-Dawley rats by adherent method.The BMSCs were identified by growth curve,flow cytometry and cell morphology observation,and the BMSCs suspension was prepared.A total of 45 Sprague-Dawley rats were randomly divided into control group,RP group and BMSC group,with 15 rats in each group.The rats in the control group were fed normally without modeling.The rats in the RP group and BMSC group were induced by sodium iodate (NaIO3) method to establish RP model.After successful modeling,the rats in the RP group were injected with 0.15 mL normal saline under the retina,and the rats in the BMSC group were injected with 0.15 mL BMSC suspension (the cell density was 1×109 L-1) under the retina,once a day for 7 days;while the rats in the control group did not take any intervention measures.The levels of serum brain-derived neurotrophic factor (BDNF) and interleukin-10 (IL-10) were detected before modeling,after modeling and one month after intervention.One month after the intervention,the rats were killed by breaking the neck,and the retinal tissues were obtained.The histological morphology of the retinal lesions was observed by hematoxylin-eosin (HE) staining.Results　Under the inverted microscope,BMSCs began to adherent growth after two hours of culture,and the pseudopodia appeared after two days of culture with different lengths;the BMSCs grew rapidly after 4 days of culture,and BMSCs with abundant cytoplasm began to form colonies;After subculture,the BMSCs were evenly distributed with slender form,and the cells were arranged in whirlpool or radial manner.The cell growth curve showed that the incubation period was 1-2 days after BMSC inoculation,and the cell growth was relatively slow;the third to fifth day after BMSC inoculation was logarithmic growth period,and the cells grew rapidly.The flow cytometry showed that the percentage of CD45+,CD29+ and CD44+ cells in the third generation of BMSC was 1.33%,79.95% and 81.49%,respectively.There was no significant difference in serum BDNF and IL-10 levels among the three groups before modeling (P>0.05).The levels of serum BDNF and IL-10 in the RP group and BMSC group were significantly lower than those in the control group after modeling (P<0.05).The levels of serum BDNF and IL-10 after modeling were significantly lower than those before modeling in the RP group and BMSC group (P<0.05).The levels of serum BDNF and IL-10 after intervention were significantly higher than those after modeling in the BMSC group (P<0.05).The levels of serum BDNF and IL-10 in the BMSC group were significantly higher than those in the RP group after intervention (P<0.05).There was no significant difference in serum BDNF and IL-10 levels among the time points of before modeling,after modeling and after intervention in the control group (P>0.05).The HE staining showed that the retinal cells of rats in the control group were well arranged and intact.The arrangement of retinal cells of rats was disordered and there were many cell fragments in the RP group.Compared with the RP group,the retinal cells of rats in the BMSC group were arranged orderly and the cell fragments were significantly reduced.Conclusion　The BMSC can significantly increase the levels of serum BDNF and IL-10 in RP rats,and promote the repair of damaged retina.

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更新日期/Last Update: 2021-06-05