早期异基因骨髓间充质干细胞局部移植修复重度碱烧伤角膜的实验研究

Research of severe alkali burned rat corneal surface by rat bone marrow-derived mesenchymal stem cells early allotrans-plantation

目的探讨结膜下异基因骨髓间充质干细胞(MMSC)移植修复重度碱烧伤角膜的可行性。方法 MMSC移植组大鼠28只(其中10只DiI标记MMSC),对照组大鼠18只。观察角膜透明度及新生血管。免疫荧光法观察角膜上皮细胞角蛋白12(CKl2)及缝隙连接蛋白43(Cx43)的表达和分布。RT-PCR分析白介素-1受体拮抗剂(IL-1RA)的表达。结果 4周时移植组角膜新生血管减少,透明度未见改善。角膜缘少量MMSC,未表达CK12、Cx43。角膜近中央区和中央区未见迁移的MMSC。移植组2周、4周IL-1RA高表达,2周时差异最明显。结论结膜下异基因MMSC移植可以抑止重度碱烧伤角膜新生血管的生成,可能与减轻炎症反应有关。

Objective To investigate the feasibility of reconstruction of severe alkali burned rat corneal surface by early subconjunctiva injection of allogenic marrow-derived mesenchymal stem cells. Methods Severe alkali burned rat corneal model was established. After aUogenetic MMSC transplantation, rat cornea clarity and neovacular in transplantation group and control group were observed. At the 4th week after DiI-labled MMSC transplantation, expression and distribution of corneal epithelial cell specific marker CK12 and Cx43 were observed by confocal microscopy. Expression of IL-IRA was compared between two groups at 3 d, 2 w and 4 w after transplantation. Results Corneas in transplantation group at the 4th week presented reduction in vascularity and no obvious improvement in clarity and edema. A lot of DiI-labled MMSC showing DAPI appeared in cornea limbus and not in central region or nearcentral region of cornea, which expressed no CK12 and Cx43. Corneas in transplantation group expressed more IL-iRA than in control group at the 2 w and 4 w, and most discrepancy at the 2 w. Conclusion Allogenetic MMSC after early being subconjunetivally transplanted into severe alkali burned rat cornea, can inhibit angiogenesis of rat corneal surface.