Gondhowiardjo T D, van Haeringen N J, Völker-Dieben H J, Beekhuis H W, Kok J H, van Rij G, Pels L, Kijlstra A
Department of Ophthalmology, University of Indonesia, Jakarta.
Cornea. 1993 Mar;12(2):146-54. doi: 10.1097/00003226-199303000-00010.
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.
在本研究中，我们使用多种电泳技术结合免疫化学和酶谱鉴定方法，研究了圆锥角膜中角膜醛脱氢酶（ALDH）的特性。将正常角膜以及角膜移植手术中获取的其他病理性角膜片用作对照。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中的酶活性显著降低（p < 0.001）（3.1±2.1 IU/mg蛋白质），而基质和内皮提取物中未观察到显著差异。在比较病理性角膜和正常对照时，未观察到角膜ALDH热稳定性行为的显著差异，54 kDa和88 kDa条带的位置也未发生明显变化。另一方面，等电聚焦分析显示病理性角膜与对照相比呈现出不同的图谱。此外，非变性聚丙烯酰胺凝胶电泳分析表明，正常角膜呈现三条带，而圆锥角膜和其他病理性角膜仅显示两条带。使用还原试剂（如谷胱甘肽）在体外可以重现从三条带模式到两条带模式的转变。
