Saghizadeh M, Brown D J, Castellon R, Chwa M, Huang G H, Ljubimova J Y, Rosenberg S, Spirin K S, Stolitenko R B, Adachi W, Kinoshita S, Murphy G, Windsor L J, Kenney M C, Ljubimov A V
Ophthalmology Research Laboratories and Neurosurgical Institute, Burns and Allen Research Institute, Cedars-Sinai Medical Center, University of California Los Angeles Medical School Affiliate, Los Angeles, California 90048, USA.
Am J Pathol. 2001 Feb;158(2):723-34. doi: 10.1016/S0002-9440(10)64015-1.
We have previously described decreased immunostaining of nidogen-1/entactin; laminin chains alpha1, alpha5, beta1,gamma1; and epithelial integrin alpha3beta1 in human diabetic retinopathy (DR) corneas. Here, using 142 human corneas, we tested whether these alterations might be caused by decreased gene expression levels or increased degradation. By semiquantitative reverse transcription-polymerase chain reaction, gene expression levels of the alpha1, alpha5, and beta1 laminin chains; nidogen-1/entactin; integrin alpha3 and beta1 chains in diabetic and DR corneal epithelium were similar to normal. Thus, the observed basement membrane and integrin changes were unlikely to occur because of a decreased synthesis. mRNA levels of matrix metalloproteinase-10 (MMP-10/stromelysin-2) were significantly elevated in DR corneal epithelium and stroma, and of MMP-3/stromelysin-1, in DR corneal stroma. No such elevation was seen in keratoconus corneas. These data were confirmed by immunostaining, zymography, and Western blotting. mRNA levels of five other proteinases and of three tissue inhibitors of MMPs were similar to normal in diabetic and DR corneal epithelium and stroma. The data suggest that alterations of laminins, nidogen-1/entactin, and epithelial integrin in DR corneas may occur because of an increased proteolytic degradation. MMP-10 overexpressed in the diabetic corneal epithelium seems to be the major contributor to the observed changes in DR corneas. Such alterations may bring about epithelial adhesive abnormalities clinically seen in diabetic corneas.
我们之前曾描述过,在人类糖尿病视网膜病变(DR)角膜中,巢蛋白-1/内动蛋白、层粘连蛋白链α1、α5、β1、γ1以及上皮整合素α3β1的免疫染色减少。在此,我们使用142个人类角膜,测试了这些改变是否可能由基因表达水平降低或降解增加所致。通过半定量逆转录-聚合酶链反应,糖尿病和DR角膜上皮中层粘连蛋白链α1、α5和β1、巢蛋白-1/内动蛋白、整合素α3和β1链的基因表达水平与正常情况相似。因此,观察到的基底膜和整合素变化不太可能是由于合成减少所致。基质金属蛋白酶-10(MMP-10/基质溶解素-2)的mRNA水平在DR角膜上皮和基质中显著升高,而MMP-3/基质溶解素-1在DR角膜基质中显著升高。圆锥角膜中未观察到这种升高。这些数据通过免疫染色、酶谱分析和蛋白质印迹法得到了证实。在糖尿病和DR角膜上皮及基质中,其他五种蛋白酶和三种MMP组织抑制剂的mRNA水平与正常情况相似。数据表明,DR角膜中层粘连蛋白、巢蛋白-1/内动蛋白和上皮整合素的改变可能是由于蛋白水解降解增加所致。糖尿病角膜上皮中过度表达的MMP-10似乎是DR角膜中观察到的变化的主要促成因素。这种改变可能导致糖尿病角膜临床上所见的上皮黏附异常。