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Sulfiredoxin-1 exerts anti-apoptotic and neuroprotective effects against oxidative stress-induced injury in rat cortical astrocytes following exposure to oxygen-glucose deprivation and hydrogen peroxide.
- Source :
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International journal of molecular medicine [Int J Mol Med] 2015 Jul; Vol. 36 (1), pp. 43-52. Date of Electronic Publication: 2015 May 08. - Publication Year :
- 2015
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Abstract
- Sulfiredoxin 1 (Srxn1), an endogenous antioxidant protein, plays an important neuroprotective role in cerebral ischemia. However, the exact mechanisms of action of Srxn1 in cerebral ischemia have not yet been fully elucidated. Therefore, in the present study, rat primary cortical astrocytes transfected with a lentiviral vector encoding short hairpin RNA (shRNA) were exposed to oxygen-glucose deprivation (OGD) for 4 h or to 100 µM hydrogen peroxide (H2O2) for 6 h, in order to construct an in vitro model of cerebral ischemia-induced damage. We found that following exposure to OGD or H2O2, the knockdown of Srxn1 resulted in a decrease in cell viability, as shown by MTS assay, an increase in cell damage, as shown by lactate dehydrogenase assay and an increase in cellular apoptosis, as shown by Hoechst 33342 staining and flow cytometry. Furthermore, we found that following exposure to OGD or H2O2, the knockdown of Srxn1 resulted in a decrease in mitochondrial transmembrane potential (Δψm) as indicated by JC-1 staining, an increase in the cytoplasmic expression of cytochrome c (Cyt.C), caspase-3, caspase-9, poly(ADP-ribose) polymerase (PARP) and Bax protein at the protein level, but a decrease in the expression of the anti-apoptotic Bcl-2 protein; these effects were tightly associated with the mitochondrial apoptotic pathway. However, we found that there was no obvious change in the intracellular calcium ([Ca2+]i) levels and caspase-12 expression following the knockdown of Srxn1. Taken together, the results from the present study demonstrate that Srxn1 protects primary rat cortical astrocytes from OGD- or H2O2-induced apoptosis and that involves the activation of the mitochondrial apoptotic pathway, which suggests that Srxn1 may be a potential target in the treatment of cerebral ischemia.
- Subjects :
- Animals
Astrocytes metabolism
Caspase 12 biosynthesis
Caspase 3 biosynthesis
Caspase 9 biosynthesis
Cell Survival
Cells, Cultured
Cerebral Cortex physiopathology
Cytochromes c biosynthesis
Glucose deficiency
Hydrogen Peroxide pharmacology
L-Lactate Dehydrogenase metabolism
Membrane Potential, Mitochondrial genetics
Membrane Potential, Mitochondrial physiology
Neuroprotective Agents metabolism
Oxidative Stress
Poly(ADP-ribose) Polymerases biosynthesis
Primary Cell Culture
Proto-Oncogene Proteins c-bcl-2 biosynthesis
RNA Interference
RNA, Small Interfering
Rats
Rats, Sprague-Dawley
bcl-2-Associated X Protein biosynthesis
Apoptosis physiology
Apoptosis Regulatory Proteins genetics
Brain Ischemia physiopathology
Cell Hypoxia physiology
Oxidoreductases Acting on Sulfur Group Donors genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1791-244X
- Volume :
- 36
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- International journal of molecular medicine
- Publication Type :
- Academic Journal
- Accession number :
- 25955519
- Full Text :
- https://doi.org/10.3892/ijmm.2015.2205