羊膜移植对单纯疱疹病毒性角膜基质炎中明胶酶活性表达的影响

Activity of Matrix Metalloproteinases in Herpetic Stromal Keratitis Mouse Model with Amniotic Membrane Transplantation

目的:探讨羊膜移植对小鼠角膜1型单纯疱疹病毒(HSV-1)感染后发生角膜基质炎(HSK)的角膜中明胶酶(Gelatinases)的活性表达变化的影响,探讨羊膜移植对HSK的治疗作用及其机制。方法:将50只BALB/c小鼠随机分为实验组和对照组,每组各25只,105pfu/L HSV-1病毒(SM44株)感染小鼠角膜,制作HSK小鼠模型。对实验组小鼠右眼角膜行羊膜移植术,对照组角膜不行羊膜移植术。在羊膜移植术后第0,2,7,14,21天处死动物取角膜。应用明胶酶谱(Gelatin Zymography)方法和计算机图像分析系统检测小鼠角膜明胶酶的酶活性表达的变化。结果:角膜感染HSV-1后,明胶酶谱显示:实验组小鼠角膜中酶原形式的pro-MMP-9(92 kU)在AMT后第2,14,21天表达增强。活性形式的MMP-9(82 kU)在AMT后各时间点均有表达,但在第2,14及21天表达增强。pro-MMP-2(72 kU)条带在AMT后各时间点均有表达。而MMP-2活性条带(62 kU)则出现在AMT后第14天和21天。对照组小鼠角膜中明胶酶的表达变化情况跟实验组类似,但是,MMP-2活性各时间点均较实验组为强(P<0.05)。而MMP-9的活性在羊膜移植后第2天(P<0.01),第14及21天(均为P<0.05)均较实验组为强。结论:羊膜移植对HSV-1感染小鼠角膜后引起的坏死性角膜基质炎形成的角膜上皮缺损和角膜基质溃疡有明显的修复作用,这种作用的机制可能是通过抑制明胶酶的活性表达而实现的。

Objective： To study the effects of amniotic membrane transplantation on the activity of gelatinases in mice with herpetic stromal keratitis （HSK）. Methods： Fifty BALB/c mice were randomly divided into two groups and infected with HSV-1 （SM44 strain）. Amniotic membrane transplantation （AMT） was perfomd in AMT group. At different time （0, 2, 7, 14 and 21 d） after AMT, the activity of gelatinase （matrix metalloproteinase-2,9） in corneas were detected by gelatin zymography, and the results were analyzed by computer medical image analysis system. Results： The gelatin zymogram showed a 92 kU band corresponding to the pro-enzyme form of MMP-9 and it was detected stronger at the 2,14 and 21 d in AMT group, the active MMP-9 （82 kU band） was present in each time point post-AMT, and （its activity） was strong at the 2nd day and even stronger at the 14th day. The pro-enzyme form of MMP-2（72 kU） was detected at each time point. A 62 kU band corresponding to the active form of MMP-2 was displayed at 14th and 21st day post-AMT. At the same time points, the similar changes were found in the levels of MMP-2 and -9 in control group, but the expression of gelatinase was greatly stronger than those in the AMT group. Conclusion.. Immediate intervention in HSK with amniotic membrane transplantation could promote epithelial healing and inhibit the development of cornea ulcerative lesions by inhibiting the activity of gelatinase.