摘要
背景已有研究证明骨髓问充质干细胞（BMSCs）移植可重建损伤的角膜上皮并促进角膜上皮的修复，但BMSCs的作用机制尚不明确。目的探讨BMSCs移植在大鼠角膜化学伤后抗炎和抗新生血管形成的作用。方法从2只成年清洁级SD大鼠双侧股骨及胫骨提取骨髓并进行BMSCs培养和传代，第3代细胞种植于羊膜。18只成年清洁级SD大鼠用NaOH滤纸片贴附于角膜中央的方法制作角膜碱烧伤模型，然后按随机数字表法随机分为3组，每组6只眼，均取右眼为模型眼。造模1周后，BMSCs移植组行带有BMSCs的羊膜移植；羊膜移植组行无BMSCs的羊膜移植；模型对照组不进行移植。术后2周裂隙灯显微镜下进行角膜透明度评分并计算角膜新生血管（CNV）面积，ELISA法检测各组大鼠角膜中白细胞介素2（IL-2）、IL-10、干扰素γ（IFN-γ）和转化生长因子β（TGF—β）的含量，荧光实时定量PCR法检测各组角膜中基质金属蛋白酶2（MMP-2）、血管内皮生长因子（VEGF）、表皮生长因子（EGF）和碱性成纤维细胞生长因子（bFGF）mRNA的表达。结果传代细胞CD90及CD29表达阳性率分别为99．78％及99．79％，CD34、CD45及C011b的表达率分别为7．90％、1．16％和1．28％，符合间充质干细胞的特征，种植于羊膜后生长状态良好。角膜碱烧伤后1周，各组大鼠角膜水肿混浊评分和CNV面积的差异均无统计学意义（F=0．021，P=0．979；F=0．076，P=0．927）。移植术后2周，羊膜移植组和BMSCs移植组大鼠角膜混浊水肿评分均明显低于模型对照组（P=0．011、0．001），CNV面积均小于模型对照组（P=0．005、0．000）。与羊膜移植组比较，BMSCs移植组CNV面积较小（P=0．001）。角膜中Th1细胞分泌的IL-2和IFNγ在BMSCs移植组明显下降，与模型对照组及羊膜移植组比较差异均有统计学意义（P=0．000、0．002、0．003、0．045），而Th2细胞分泌的IL
[Abstract] Background The favorable effect of bone marrow mesenchymal stem cells （BMSCs） on the reconstruction of injured corneas have been reported, but the mechanism remains unclear. Objective This study was to evaluate the anti-inflammatory and antiangiogenic effect after transplantation of BMSCs in chemically burned corneas. Methods BMSCs were isolated and extracted from the bone marrow. The cells were cultured and passaged and then were seeded on the amniotic membrane. Corneal alkali injury models were created in 18 clean SD rats by sticking the filter paper containing 1 mol/L NaOH at the central cornea for 40 seconds. The rats were then randomized into 3 groups. Amniotic membrane with BMSCs or amniotic membrane without BMSCs were transplanted in 1 week after the establishment of models, and the rats without transplantation were used as the control group. The severity of corneal lesion was graded, and angiogenesis area was measured 2 weeks after the transplantation. Theexpression of interleukin-2 （ IL-2 ）, interferon-γ （ IFN-γ）, IL-10 and transforming growth factor-β （ TGF-β ） were examined by ELISA, and the mRNA of the matrix metalloproteinase-2 （ MMP-2 ） , vascular endothelial growth factor （VEGF） ,epidermal growth factor（EGF） and basic fibroblast growth factor（bFGF） were analyzed by real-time PCR. Results The positive rates of the cells were 99.78% and 99.79% for CD90 and CD29,7.90% , 1.16% and 1.28% for CD34,CD45 and CDllba. The ceils grew well on the amniotic membrane. The corneal inflammatory score and neovascularization area were similar among the three groups （ F = 0. 021, P = 0. 979 ; F = 0. 076, P = 0. 927 ）. However, the corneal inflammatory score was significantly reduced and neovascularization area was significantly less in the amniotic membrane group compared with the BMSCs group and control group（ P=0. 011,0. 001 ;P = 0. 005,0. 000 ）. The levels of IL-2 and IFN-γ secreted by Thl cells were decreased （ P = 0. 000,0. 002 ; P = 0. 003,0. 045 ） and
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2012年第3期253-257,共5页
Chinese Journal Of Experimental Ophthalmology
基金
南通市社会发展基金项目（$2009029）
关键词
骨髓间充质干细胞
化学烧伤
角膜
炎症相关因子
血管生成相关因子
Bone marrow mesenchymal stem cell
Chemical burn
Cornea
Inflammatory-related factor
Angiogenic-related factor
