[1]吴利安,王从毅,杨新光,等. 骨髓间充质干细胞移植对兔碱烧伤角膜新生血管形成的抑制作用[J].眼科新进展,2015,35(7):623-625.[doi:10.13389/j.cnki.rao.2015.0169]
　WU Li-An,WANG Cong-Yi,YANG Xin-Guang,et al. Inhibitive effects of bone marrow stem cells on corneal neovascularization in rabbit with corneal alkali burn[J].Recent Advances in Ophthalmology,2015,35(7):623-625.[doi:10.13389/j.cnki.rao.2015.0169]
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 骨髓间充质干细胞移植对兔碱烧伤角膜新生血管形成的抑制作用
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年7期
页码:
623-625
栏目:
实验研究
出版日期:
2015-07-05

文章信息/Info

Title:
 Inhibitive effects of bone marrow stem cells on corneal neovascularization in rabbit with corneal alkali burn
作者:
 吴利安; 王从毅; 杨新光; 王建洲; 张林; 范晶晶
 710004　陕西省西安市，西安市第四医院，西安交通大学医学院附属广仁医院
Author(s):
 WU Li-An;  WANG Cong-Yi;  YANG Xin-Guang;  WANG Jian-Zhou;  ZHANG Lin; FAN Jing-Jing
 Xi ’an Fourth Hospital, Guangren Hospital Affiliated to Xi ’ an Jiaotong University, Xi ’ an 710004 , Shaanxi Province , China
关键词:
 ; 骨髓间充质干细胞; 角膜碱烧伤; 基质金属蛋白酶-2; 肿瘤坏死因子-α
Keywords:
 ; bone marrow stem cell; corneal alkali burn ;  MMP-2 ; TNF-α
DOI:
10.13389/j.cnki.rao.2015.0169
文献标志码:
A
摘要:
 目的　观察角膜碱烧伤兔行骨髓间充质干细胞（ｂｏｎｅｍａｒｒｏｗｍｅｓｅｎｃｈｙｍａｌｓｔｅｍｃｅｌｌｓ，ＢＭＳＣ）移植后不同时间受损角膜新生血管（ｃｏｒｎｅａｌｎｅｏｖａｓｃｕｌａｒｉｚａｔｉｏｎ，ＣＮＶ）发生情况，探讨ＢＭＳＣ移植后对碱烧伤ＣＮＶ的抑制作用。方法　取日本大白兔２４只，随机分为实验组和对照组各１２只，实验组进行烧伤后ＢＭＳＣ移植，体外分离培养得到ＢＭＳＣ，制备中度角膜碱烧伤模型，随后行ＢＭＳＣ移植。对照组不进行移植。分别于移植后３ｄ、１４ｄ、２８ｄ观察ＣＮＶ生长状态、计算ＣＮＶ面积、采用ＲＴ-ＰＣＲ检测角膜中基质金属蛋白酶-２（ＭＭＰ-２）和肿瘤坏死因子α（ＴＮＦ-α）的表达情况。结果　ＢＭＳＣ贴壁生长，呈长梭形，ＣＤ９０呈现高表达（表达率９７．９４％），ＣＤ３１低表达（表达率０．１５％）。ＢＭＳＣ移植后２８ｄ时实验组角膜较对照组明显透亮，新生血管面积显著减小（Ｐ＜０．０５）。ＢＭＳＣ移植后１４ｄ、２８ｄ，实验组ＭＭＰ-２基因条带的灰度值均低于对照组（均为Ｐ＜０．０５），实验组ＭＭＰ-２ｍＲＮＡ表达低于对照组且１４ｄ时差异最明显（Ｐ＜０．０１）。ＢＭＳＣ移植后３ｄ，实验组ＴＮＦ-α基因条带灰度值较对照组低，实验组ＴＮＦ-αｍＲＮＡ的表达明显受到抑制，且两组间差异有统计学意义（Ｐ＜０．０５）；１４ｄ时，实验组ＴＮＦ-α基因表达仍低于对照组（Ｐ＜０．０５），２８ｄ时实验组与对照组ＴＮＦ-αｍＲＮＡ表达量相当，差异无统计学意义（Ｐ＞０．０５）。结论　ＢＭＳＣ通过抑制ＭＭＰ-２的表达，减少细胞外基质的溶解，抑制ＣＮＶ的形成；通过降低炎性细胞因子ＴＮＦ-α的表达，减轻局部炎症反应，从而抑制ＣＮＶ的生长。
Abstract:
 Objective To observe the corneal neovascularization ( CNV) of damaged cornea in different time after early transplantation of autologous bone marrow stem cells ( BMSC ) in rabbits with corneal alkali burn . and explore the inhibitive effects of BMSC on the corneal alkali burn of rabbits. Methods Twenty-four rabbits were randomly divided into experimental group and control group, 12 cases in each group. BMSC was isolated and cultured in vitro. The models of moderate corneal alkali bum were induced. and the BMSC was transplanted in the experimental group. CNV growth was observed at 3 days, 14 days, 28 days after transplantation, and the CNV area was calculated. RT-PCR was used to exam the expression of MMP-2 and TNF-α mRNA in the damaged cornea. Results BMSC cultured in vitro adhered to the bottle wall, exhibiting spindle-shaped morphology , and with high expression of CD90 ( the expressive rate was 97. 94% ) ,low expression of CD31 ( the expressive rate was 0. 15% ). On the 28th day after transplantation.the degrees of corneal opacity in the experimental group was low . and CNV area in the experimental group were lower than that in the control group (P < 0. 05) . On the 14th day,28th day after transplantation , the MMP-2 gray value of the gene band in the experimental group were lower than those in the control group ( all P <0. 05 ) ,especially on the 14th day (P < 0. 01). On the 3th day after transplantation, the TNF-α gray value of the gene band in the experimental group were lower than that of the control group (P < 0. 05 ) . On the 14th day , the expression of TNF-a in the experimental group were lower than that of the control group ( P < 0. 05 ) .but on the 28 th day , there was no statistical difference between two groups (P > 0. 05) . Conclusion BMSC can inhibit the CNV through inhibiting the expression of MMP-2 and reducing the dissolution of the extracellular matrix ; BMSC can inhibit the CNV development through reducing the expression of inflammatory cytokines TNF-α and reducing the release of inflammatory proteins.

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备注/Memo

备注/Memo:
 陕西省自然科学基金资助（编号：２０１０ＪＱ４０１５）
更新日期/Last Update: 2015-07-03