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Expression of glutathione transferases in corneal cell lines, corneal tissues and a human cornea construct.
- Source :
-
International journal of pharmaceutics [Int J Pharm] 2016 Jun 15; Vol. 506 (1-2), pp. 371-81. Date of Electronic Publication: 2016 Apr 22. - Publication Year :
- 2016
-
Abstract
- 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.<br /> (Copyright © 2016 Elsevier B.V. All rights reserved.)
- Subjects :
- Animals
Blotting, Western
Cell Line
Ethacrynic Acid metabolism
Fluorescent Antibody Technique
Glutathione analogs & derivatives
Glutathione metabolism
Humans
Rabbits
Species Specificity
Swine
Cornea enzymology
Epithelium, Corneal enzymology
Glutathione S-Transferase pi metabolism
Glutathione Transferase metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1873-3476
- Volume :
- 506
- Issue :
- 1-2
- Database :
- MEDLINE
- Journal :
- International journal of pharmaceutics
- Publication Type :
- Academic Journal
- Accession number :
- 27113863
- Full Text :
- https://doi.org/10.1016/j.ijpharm.2016.04.053