肿瘤坏死因子-α对体外培养的兔角膜上皮细胞基质金属蛋白酶表达活性的影响

Effect of Tumor Necrosis Factor-Alpha on Metalloproteinases Activity of Cultured Rabbit Corneal Epithelial Cells

目的:观察肿瘤坏死因子-α(TNF-α)对体外培养的兔角膜上皮细胞基质金属蛋白酶-2、-9(MMP-2、MMP-9)表达活性的影响,探讨MMP-2、MMP-9的调节机制及TNF-α和MMP-2、MMP-9在角膜炎症性病变中的作用。方法:组织块法培养兔角膜上皮细胞,经不同浓度(1,10,100μg/L)TNF-α诱导24 h后,收集条件培养基应用明胶酶谱法检测MMP-2、MMP-9的活性,利用计算机图像分析系统进行统计分析。结果:在体外培养的角膜上皮细胞中,TNF-α能增强MMP-2、MMP-9的活性,并具有一定的浓度依赖性,TNF-α可激活MMP-2酶原,使其表现出活化形式。结论:炎性因子TNF-α可以上调角膜上皮细胞分泌的MMP-2、MMP-9的活性,针对这一过程进行干预对防治角膜炎性疾病具有重要意义。

Objective： To study the activity of matrix metalloproteinase-2, -9 （MMP-2, MM-9） genera- ted by the cultured rabbit corneal epithelial cells stimulated by tumor necrosis factor-α（TNF-α）, and to investigate the possible regulative mechanism of MMP-2 and MMP-9, and the potential effect in the corneal inflammatory diseases. Methods： The rabbit corneal epithelium cells were cultured in vitro and incubated with TNF-α at different concentrations （0, 1, 10, 100 μg/L） for 24 h. The activity of Pro-MMP-2 and MMP-9 was examined by using gelatin zymography. The results were analyzed by the computer image-analysis system and statistical test. Results. TNF-α stimulated the secretion of MMP-2 and MMP-9 in a dose dependent manner. Pro-MMP-2 was ac- tivated by TNF-α to show its activated form. Conclusion： Inflammatory factor such as TNF-α can stimulate MMP-2 and＇MMP-9 activity of corneal epithelial cells, and this may be one manipula- ting mechanisms of MMPs expression in the pathogenesis of corneal diseases, which may play an important role in the prevention and treatment of corneal inflammatory diseases.