ANALYSIS OF CORNEAL ALDEHYDE DEHYDROGENASE PATTERNS IN PATHOLOGICAL CORNEAS
ANALYSIS OF CORNEAL ALDEHYDE DEHYDROGENASE PATTERNS IN PATHOLOGICAL CORNEAS
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DOI：
10.1097/00003226-199303000-00010
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发表时间：
1993-03-01
期刊：
影响因子：
2.8
通讯作者：
KIJLSTRA, A
中科院分区：
文献类型：
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作者：
GONDHOWIARDJO, TD;VANHAERINGEN, NJ;KIJLSTRA, A
在本研究中，我们使用各种电泳技术结合免疫化学和酶谱鉴定，研究了圆锥角膜中角膜醛脱氢酶（ALDH）的特性。使用在角膜整形手术期间获得的正常角膜和其他病理性角膜纽扣作为对照。与正常对照（5.5 +/- 2.6 IU/mg 蛋白质）相比，圆锥角膜上皮提取物（3.1 +/- 2.1 IU/mg 蛋白质）的酶活性显着降低（p < 0.001），而基质和内皮提取物中未观察到显着差异。与病理角膜和正常对照相比，角膜 ALDH 热稳定性行为没有观察到显着差异，54-和 88-kDa 物质的位置也没有发生任何明显变化。另一方面，等电聚焦分析显示病理角膜与对照相比有不同的模式。此外，天然聚丙烯酰胺凝胶电泳分析显示，正常角膜显示出三个条带，而圆锥角膜和其他病理角膜仅显示出两个条带。从三带模式到两带模式的转变可以使用谷胱甘肽等还原剂在体外重现。
In this study we investigated the properties of corneal aldehyde dehydrogenase (ALDH) in keratoconus corneas using various electrophoretic techniques combined with immunochemical and zymographic identification. Normal corneas and other pathologic corneal buttons obtained during keratoplastic surgery were used as a control. A significant (p < 0.001) lower enzymatic activity was found in keratoconus epithelial extracts (3.1 +/- 2.1 IU/mg protein) compared with normal controls (5.5 +/- 2.6 IU/mg protein), whereas no significant differences were observed in the stromal and endothelial extracts. No significant differences were observed for the corneal ALDH thermolability behavior, nor did any marked changes occur in the position of the 54- and 88-kDa species when comparing the pathologic corneas and normal controls. On the other hand, isoelectric focusing analysis showed a different pattern for the pathologic corneas as compared with controls. Moreover, native-polyacrylamide gel electrophoresis analysis showed that normal corneas exhibit three bands, whereas keratoconus and other pathologic corneas only show two bands. The shift from the three-band pattern to the two-band pattern could be reproduced in vitro using reducing agents, such as glutathione.