不同浓度重组Canstatin蛋白对碱烧伤后角膜MMP-2及TIMP-2表达的影响  

Effect of recombinant canstatin proteins with different concentrations on the expression of matrix metaiioproteinase-2 and tissue inhibitor of metalloproteinase-2 in mice with corneal alkaline burn

作　　者：鲁铭[1] 朱晶[1] 

机构地区：[1]中国湖北省武汉市第一医院眼科,430022

出　　处：《国际眼科杂志》2016年第2期224-227,共4页International Eye Science

摘　　要：目的:探讨不同浓度重组Canstatin蛋白对碱烧伤后小鼠角膜基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)及其组织抑制剂-2(tissue inhibitor of metalloproteinase-2,TIMP-2)表达的影响及其调节作用。方法:BALB/c小鼠60只随机分为实验组A、实验B及对照组C,每组20只。采用1mol/L氢氧化钠溶液烧伤小鼠右眼角膜,建立炎症性角膜碱烧伤动物模型,分别予以A组、B组重组Canstatin蛋白3μg/mL、5μg/mL点右眼,4次/d,对照组C组予以生理眼水点右眼。在碱烧伤后第1、3、7、14d以形态学分析评价角膜上皮损伤面积及新生血管生长的情况,并于碱烧伤后第1、3、7、14d应用Western-blot检测角膜MMP-2和TIMP-2的表达,增强化学发光法(ECL)对结果进行分析。结果:形态学分析显示,A组和B组小鼠在碱烧伤后第3d起各时间点角膜上皮缺损面积均小于对照组(P<0.01),角膜新生血管均得到抑制,CNV面积明显小于对照组(P<0.01)。Western-blot结果显示,碱烧伤后各时间点MMP-2的表达,A组和B组均明显低于对照组(P<0.01),TIMP-2的表达高于对照组(P<0.01),且A组和B组间MMP-2的表达在第14d比较差异有统计学意义(P<0.05),TIMP-2的表达在第7d及第14d比较差异有统计学意义(P<0.05)。结论:重组Canstatin蛋白可通过抑制角膜细胞及浸润的炎性细胞产生MMP-2,促进TIMP-2表达,从而抑制和延迟碱烧伤后角膜融解的发生和发展,对碱烧伤后角膜的重塑起着重要作用。AIM：To investigate the effect of recombinant canstatin proteins with different concentration on the expressions of matrix metalloproteinase-2（MMP-2） and tissue inhibitor of metalloproteinase-2（TIMP-2） in mice with corneal alkali burn.METHODS：Sixty BALB/c mice were divided into three groups（experimental group A,experimental group B and control group C）,20 mice in every group and their corneas in the right eyes were burned with alkali（1mol/L NaOH）.The experimental group A received recombinant canstatin proteins drops with 3μg/mL.The experimental group B received recombinant canstatin proteins drops with 5μg/mL and the control group C was treated with physiologic saline.At different time points（1,3,7 and 14d） after alkali burns,the mice were killed and the growth of epithelial defect and corneal neovascularization（CNV）were observed with an operation microscope.The expressions of MMP-2 and TIMP-2 in cornea were measured by the Western blot technique,and the results were analyzed by enhanced chemiluminescent（ECL）.RESULTS：The areas of epithelial defect and corneal neovasularization significantly reduced in mice treated with recombinant canstatin proteins compared to mice treated with phvsioloaic saline at 3,7 and 14 d after alkaliinduced injury（all P〈0.01）;the neovasularization was suppressed and the area of CNV was less than that in control group C（all P〈0.01）.Western blot analysis showed that the expression levels of MMP- 2 in experimental group A and B were significantly lower than that in control group C（P〈0.01） and the expressions of TIMP-2 in experimental group A and B were significantly higher（P〈0.01）;the level of MMP-2 in experimental group B were lower than that in experimental group A on day 14（P 0.05）,while the level of TIMP- 2 in experimental group B were significantly higher than that in experimental group A on day 7 and day 14（P〈0.05）.CONCLUSION：Recombinant canstatin proteins may suppress the expression of MMP-2,upregulate the express

关 键 词：碱烧伤 重组Canstatin蛋白 基质金属蛋白酶-2 基质金属蛋白酶组织抑制剂-2 

分 类 号：R779.13[医药卫生—眼科]