羊膜对角膜成纤维细胞CTGF表达及凋亡的影响  

Effects of amniotic membrane on expression of connective tissue growth factor and apoptosis of corneal fibroblasts

作　　者：黄琼[1] 胡燕华[1] 张明昌[1] 张缨[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,武汉430022

出　　处：《眼科研究》2006年第2期125-128,共4页Chinese Ophthalmic Research

摘　　要：目的观察羊膜对在其基质面生长的角膜成纤维细胞结缔组织生长因子(CTGF)表达以及凋亡的影响。方法将传代的家兔角膜成纤维细胞接种于羊膜基质面,培养5d后,用RT-PCR检测角膜成纤维细胞CTGFmRNA表达的变化;用流式细胞仪观察羊膜对IL-1诱导的角膜成纤维细胞凋亡的影响。结果在羊膜基质面培养的角膜成纤维细胞,其CTGF/GAPDH像素比值较对照组低,与对照组比较有明显统计学差异(P<0.05);羊膜能降低IL-1诱导的角膜成纤维细胞凋亡率,与对照组比较有明显统计学差异(P<0.01)。结论羊膜不仅抑制体外培养的角膜成纤维细胞CTGFmRNA的表达,而且能抑制角膜成纤维细胞的凋亡,可能部分解释羊膜减轻角膜基质损伤修复反应和抗眼表瘢痕形成的机制。Objective Amniotic membrane has been shown the ability to reduce ocular surface scar formation. The aim of this study is to investigate the effects of amniotic membrane on expression of connective tissue growth factor （CTGF） and apoptosis of corneal fibroblasts cultured on the stroma of amniotic membrane. Methods The ceils of the control group and amniotic membrane group were both seeded in DMEM containing 10% fetal bovine serum,while the cells in the latter group were cultured on amniotic membrane stroma. After five days of culture,reverse transcription polymerase-chain reaction （RT-PCR） was used to detect CTGF mRNA expression, Interleukin-1 （IL-1） （10 ng/ml） was used to induce cell apoptosis,and the cells from both groups were seeded in DMEM containing 2% fetal bovine serum, while the cells in the amniotic membrane group were cultured on amniotic membrane stroma. After five days of induction, flow cytometry was used to evaluate the effect of amniotic membrane on corneal fibroblasts apoptosis induced by IL-1. Results The CTGF/GAPDH value in corneal fibroblasts on the stroma of amniotic membrane was 0.96±0.087 and that in normal cultured corneal fibroblasts was 1.37±0.121 and the difference between them was significant （P 〈 0.05）, suggesting amniotic membrane could suppress CTGF expression in cultured corneal fibroblasts. The percent of apoptosis in the amniotic membrane group （6.32±0.73） was considerably lower than that in the control group （12.98±1.85）（P 〈 0.01）, showing amniotic membrane could protect corneal fibroblasts from apoptosis. Conclusion Amniotic membrane significantly inhibit corneal fibroblasts apoptosis induced by IL-1, which may partly explain why amniotic membrane could reduce the tissue reaction during corneal wound healing and inhibit ocular surface scar formation.

关 键 词：羊膜 角膜成纤维细胞 结缔组织生长因子 凋亡 

分 类 号：R77[医药卫生—眼科]