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Expression of antioxidant enzymes in rat retinal ischemia followed by reperfusion.

Agardh, Carl-David LU ; Gustavsson, Carin LU ; Hagert, Per LU ; Bengtsson, Marie E LU and Agardh, Elisabet LU (2006) In Metabolism, Clinical and Experimental 55(7). p.892-898
Abstract
To evaluate the expression and protein levels of antioxidant enzymes in the rat retina exposed to oxidative stress induced by ischemia-reperfusion injury. Retinal ischemia was induced in female Wistar rats by ligation of the optic nerve and vessels behind the left eye bulb, and was followed by reperfusion for 0, 3, 6, or 24 hours. The right eye served as control. RNA and protein were extracted simultaneously from each retina. Expressions of the endogenous antioxidant enzymes glutathione peroxidase (GPx1), catalase (CAT), copper/zinc superoxide dismutase, manganese superoxide dismutase, and the catalytic subunit of glutamylcysteine ligase (GCLc) were analyzed with real-time reverse transcription polymerase chain reaction and related to the... (More)
To evaluate the expression and protein levels of antioxidant enzymes in the rat retina exposed to oxidative stress induced by ischemia-reperfusion injury. Retinal ischemia was induced in female Wistar rats by ligation of the optic nerve and vessels behind the left eye bulb, and was followed by reperfusion for 0, 3, 6, or 24 hours. The right eye served as control. RNA and protein were extracted simultaneously from each retina. Expressions of the endogenous antioxidant enzymes glutathione peroxidase (GPx1), catalase (CAT), copper/zinc superoxide dismutase, manganese superoxide dismutase, and the catalytic subunit of glutamylcysteine ligase (GCLc) were analyzed with real-time reverse transcription polymerase chain reaction and related to the endogenous control cyclophilin B. Protein levels were measured with Western blot analysis. During the early phase (0 or 3 hours) of reperfusion, no changes were seen in enzyme expression. After 6 hours, GCLc expression increased by a factor of 1.14 (P =.034), followed by a decline of 0.80 after 24 hours (P =.00004), according to the comparative Ct method. After 24 hours of reperfusion, GPx1 expression increased by a factor of 1.14 (P =.028), and CAT had decreased by 0.82 (P =.022). Expressions of copper/zinc superoxide dismutase and manganese superoxide dismutase showed a tendency toward a decrease by factors of 0.86 (P =.055) and 0.88 (P =.053), respectively, after 24 hours. Protein levels did not differ for any of the antioxidants, regardless of reperfusion time. The slightly increased messenger RNA expression of GPx1 after 24 hours of reperfusion with a concomitant very modest decrease in CAT and GCLc expression and no change in protein levels indicate a very modest, if any, response to oxidative stress generated by ischemia followed by reperfusion in rat retina. (Less)
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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Metabolism, Clinical and Experimental
volume
55
issue
7
pages
892 - 898
publisher
Elsevier
external identifiers
  • pmid:16784960
  • wos:000238691700007
  • scopus:33745133331
  • pmid:16784960
ISSN
1532-8600
DOI
10.1016/j.metabol.2006.02.016
language
English
LU publication?
yes
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Department of Clinical Sciences, Malmö (013240000), Unit on Vascular Diabetic Complications (013241510), Ophthalmology (013242810), Medical Genetics Unit (013241550)
id
a02b1aa1-6c3d-4e98-864c-9f6bb95df898 (old id 158235)
date added to LUP
2016-04-01 16:06:14
date last changed
2021-05-19 01:33:35
@article{a02b1aa1-6c3d-4e98-864c-9f6bb95df898,
  abstract     = {To evaluate the expression and protein levels of antioxidant enzymes in the rat retina exposed to oxidative stress induced by ischemia-reperfusion injury. Retinal ischemia was induced in female Wistar rats by ligation of the optic nerve and vessels behind the left eye bulb, and was followed by reperfusion for 0, 3, 6, or 24 hours. The right eye served as control. RNA and protein were extracted simultaneously from each retina. Expressions of the endogenous antioxidant enzymes glutathione peroxidase (GPx1), catalase (CAT), copper/zinc superoxide dismutase, manganese superoxide dismutase, and the catalytic subunit of glutamylcysteine ligase (GCLc) were analyzed with real-time reverse transcription polymerase chain reaction and related to the endogenous control cyclophilin B. Protein levels were measured with Western blot analysis. During the early phase (0 or 3 hours) of reperfusion, no changes were seen in enzyme expression. After 6 hours, GCLc expression increased by a factor of 1.14 (P =.034), followed by a decline of 0.80 after 24 hours (P =.00004), according to the comparative Ct method. After 24 hours of reperfusion, GPx1 expression increased by a factor of 1.14 (P =.028), and CAT had decreased by 0.82 (P =.022). Expressions of copper/zinc superoxide dismutase and manganese superoxide dismutase showed a tendency toward a decrease by factors of 0.86 (P =.055) and 0.88 (P =.053), respectively, after 24 hours. Protein levels did not differ for any of the antioxidants, regardless of reperfusion time. The slightly increased messenger RNA expression of GPx1 after 24 hours of reperfusion with a concomitant very modest decrease in CAT and GCLc expression and no change in protein levels indicate a very modest, if any, response to oxidative stress generated by ischemia followed by reperfusion in rat retina.},
  author       = {Agardh, Carl-David and Gustavsson, Carin and Hagert, Per and Bengtsson, Marie E and Agardh, Elisabet},
  issn         = {1532-8600},
  language     = {eng},
  number       = {7},
  pages        = {892--898},
  publisher    = {Elsevier},
  series       = {Metabolism, Clinical and Experimental},
  title        = {Expression of antioxidant enzymes in rat retinal ischemia followed by reperfusion.},
  url          = {https://lup.lub.lu.se/search/files/4568638/625497.pdf},
  doi          = {10.1016/j.metabol.2006.02.016},
  volume       = {55},
  year         = {2006},
}