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Ula V. Jurkunas, Ian Rawe, Maya S. Bitar, Cheng Zhu, Deshea L. Harris, Kathryn Colby, Nancy C. Joyce; Decreased Expression of Peroxiredoxins in Fuchs’ Endothelial Dystrophy. Invest. Ophthalmol. Vis. Sci. 2008;49(7):2956-2963. doi: 10.1167/iovs.07-1529.
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purpose. To compare the relative expression of peroxiredoxin (Prx) proteins in normal human corneal endothelium with endothelium in corneas affected by Fuchs’ endothelial dystrophy (FED) and between normal human endothelium and epithelial/stromal tissue.
methods. Human corneal endothelial cell-Descemet’s membrane (HCEC-DM) complexes from normal and FED corneal buttons were dissected from the epithelium/stroma. For proteomic analysis, HCEC-DM protein extracts were separated by using two-dimensional gel electrophoresis. Relative differences in protein spot density was analyzed. Proteins of interest, including Prx isoforms, were identified by MALDI-TOF (matrix-assisted desorption ionization-time of flight) mass spectrometry. Western blot analysis compared the relative expression of Prx isoforms in normal and FED endothelium and between normal endothelium and normal epithelium/stroma. Expression of Prx-2 mRNA was compared by using real-time PCR.
results. Proteomic analysis identified differences in the relative expression of Prx isoforms between normal and FED endothelium. Western blot analysis confirmed that expression of Prx-2, −3, and −5 was significantly decreased (P < 0.05) in FED cells. Normal HCECs expressed significantly (P < 0.05) higher levels of Prx-2 and −3 than did the epithelium/stroma. Expression of Prx-5 was not significantly different (P > 0.05) in the endothelium versus the epithelium/stroma. Real-time PCR analysis revealed that Prx-2 mRNA was significantly decreased (P = 0.027) in FED samples.
conclusions. Prx proteins were identified in human corneal endothelium. The fact that Prx-2 and −3 were expressed at significantly higher levels in HCEC-DM compared with the epithelium/stroma reflects the different physiologic activities of individual corneal cell types. Significantly decreased expression of Prx-2, −3, and −5 in FED may suggest an alteration in the ability of endothelial cells to withstand oxidant-induced damage and may be closely related to the pathogenesis of this disease.
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