Expression of glutathione transferases in corneal cell lines, corneal tissues and a human cornea construct

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• 作者：Christian Kö ; lln ^{c.koelln@tu-braunschweig.de" class="auth_mail" title="E-mail the corresponding author} ; Stephan Reichl ; ^{s.reichl@tu-braunschweig.de" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Cornea ; Cell culture in vitro model ; Metabolism ; Glutathione transferase ; GSTP1-1 ; GSTO1-1
• 刊名：International Journal of Pharmaceutics
• 出版年：2016
• 出版时间：15 June 2016
• 年：2016
• 卷：506
• 期：1-2
• 页码：371-381
• 全文大小：1230 K

文摘

Glutathione transferase (GST) expression and activity were examined in a three-dimensional human cornea construct and were compared to those of excised animal corneas. The objective of this study was to characterize phase II enzyme expression in the cornea construct with respect to its utility as an alternative to animal cornea models. The expression of the GSTO1-1 and GSTP1-1 enzymes was investigated using immunofluorescence staining and western blotting. The level of total glutathione transferase activity was determined using 1-chloro-2,4- dinitrobenzene as the substrate. Furthermore, the levels of GSTO1-1 and GSTP1-1 activity were examined using S-(4-nitrophenacyl)glutathione and ethacrynic acid, respectively, as the specific substrates. The expression and activity levels of these enzymes were examined in the epithelium, stroma and endothelium, the three main cellular layers of the cornea. In summary, the investigated enzymes were detected at both the protein and functional levels in the cornea construct and the excised animal corneas. However, the enzymatic activity levels of the human cornea construct were lower than those of the animal corneas.