目的：探讨早产儿视网膜病变（ROP）大鼠玻璃体内注射骨髓间充质干细胞（BMSCs）对视网膜细胞凋亡及视网膜中神经营养素-3（NT-3）、睫状神经营养因子（CNTF）表达的影响。方法：40只7 日龄 Sprague-Dawley新生大鼠随机分为正常对照组、ROP模型组、骨髓间充质干细胞（BMSCs）移植组与磷酸缓冲液组（PBS组），每组10只。采用高氧法制成 ROP 模型，持续高氧5 d后分别给予BMSCs移植组与PBS组玻璃体内注射BMSCs和PBS。移植7 d后，采用 TUNEL/DAPI、NT-3/DAPI、CNTF/DAPI免疫荧光双标法观察BMSCs移植对视网膜细胞凋亡及NT-3、CNTF表达的影响。结果：移植7 d后，正常对照组视网膜几乎未见TUNEL+ DAPI+细胞，BMSCs移植组视网膜可见少量TUNEL+ DAPI+细胞，显著低于ROP模型组与PBS组（P0.05）；正常对照组视网膜可见极少量的NT-3+DAPI+与CNTF+DAPI+ 细胞，ROP模型组与PBS组大鼠视网膜可见少量NT-3+DAPI+与CNTF+DAPI+ 细胞，较正常对照组增多，但差异无统计学意义（P>0.05）；BMSCs移植组大鼠视网膜NT-3+DAPI+与CNTF+DAPI+ 细胞数均显著多于ROP模型组与PBS组(P0.05）。结论：BMSCs移植可减轻ROP大鼠视网膜细胞的凋亡，其机制可能与其促进ROP大鼠视网膜细胞NT-3与CNTF的表达有关。
Abstract
OBJECTIVE: To explore the effects of marrow mesenchymal stem cell (BMSC) transplantation on retinal cells apoptosis and changes to neurotrophin-3 (NT-3 and ciliary neurotrophic factor (CNTF) in rats with retinopathy of prematurity (ROP). METHODS: Seven-day-old Sprague-Dawley rats were randomly divided into normal control (CON), ROP, BMSC transplantation (BMSCs were transplanted 5 days after oxygen conditioning) and phosphate buffered saline (PBS) groups. The ROP model was prepared according to the classic hyperoxygen method. Seven days after transplantation, TUNEL/DAPI, NT-3/API and CNTF/DAPI double-labeled immunofluorescence were used to examine the effects of BMSC transplantation on both the apoptosis of retinal cells and the expression of NT-3 and CNTF protein in the retinal cells of the ROP rats. RESULTS: Seven days after BMSC transplantation, there were few TUNEL+ DAPI+ cells observed in the CON group. There were fewer TUNEL+DAPI+ cells observed in the BMSC group than in the ROP group (P0.05). There were few NT-3+DAPI+ cells and CNTF+DAPI+ cells in the CON group. There were more NT-3+DAPI+ and CNTF+DAPI+ cells in the ROP group than in the CON group, but there was no significant difference between the ROP and CON groups (P>0.05). More NT-3+DAPI+ and CNTF+DAPI+ cells were observed in the BMSC group compared with the ROP group (P0.05). CONCLUSIONS: BMSC transplantation therapy could alleviate the apoptosis of retinal cells in ROP rats, and its mechanisms might be associated with promoting the expression of NT-3 and CNTF protein in retinal cells.
关键词
视网膜病变 /
骨髓间充质干细胞移植 /
凋亡 /
神经营养素-3 /
睫状神经营养因子 /
大鼠
Key words
Retinopathy /
Bone marrow mesenchymal stem cell transplantation /
Apoptosis /
Neurotrophin-3 /
Ciliary neurotrophic factor /
Rats
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