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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.

Zhou Y, Zhou Y, Yu S, Wu J, Chen Y, Zhao Y - Int. J. Mol. Med. (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Chongqing Medical University, Chongqing 400016, P.R. China.

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.

No MeSH data available.


Related in: MedlinePlus

Knockdown of sulfiredoxin 1 (Srxn1) decreases cell viability and increases cell damage following exposure to oxygen-glucose deprivation (OGD) or hydrogen peroxide (H2O2). (A) Cell viability was determined by MTS assay. Exposure to OGD or H2O2 decreased cell viability. Following exposure to OGD or H2O2, the knockdown of Srxn1 promoted the reduction in cell viability. Values are the means ± SEM; *P<0.05, **P<0.01 vs. controls (untreated cells); #P<0.05 vs. control + OGD (or H2O2) group. n=6 from 3 independent experiments. (B) Cell damage was assessed by measuring the lactate dehydrogenase (LDH) levels. Exposure to OGD or H2O2 increased cell damage. Following exposure to OGD or H2O2, the knockdown of Srxn1 increased cell damage even more significantly. n=3 from 3 independent experiments. Values are the means ± SEM; **P<0.01 vs. controls (untreated cells); ##P<0.01 vs. control + OGD (or H2O2) group. NC, negative control.
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f2-ijmm-36-01-0043: Knockdown of sulfiredoxin 1 (Srxn1) decreases cell viability and increases cell damage following exposure to oxygen-glucose deprivation (OGD) or hydrogen peroxide (H2O2). (A) Cell viability was determined by MTS assay. Exposure to OGD or H2O2 decreased cell viability. Following exposure to OGD or H2O2, the knockdown of Srxn1 promoted the reduction in cell viability. Values are the means ± SEM; *P<0.05, **P<0.01 vs. controls (untreated cells); #P<0.05 vs. control + OGD (or H2O2) group. n=6 from 3 independent experiments. (B) Cell damage was assessed by measuring the lactate dehydrogenase (LDH) levels. Exposure to OGD or H2O2 increased cell damage. Following exposure to OGD or H2O2, the knockdown of Srxn1 increased cell damage even more significantly. n=3 from 3 independent experiments. Values are the means ± SEM; **P<0.01 vs. controls (untreated cells); ##P<0.01 vs. control + OGD (or H2O2) group. NC, negative control.

Mentions: Following exposure to OGD or H2O2, cell viability was determined by MTS assay (Fig. 2A). Compared with the control group, following exposure to OGD or H2O2, cell viability was decreased by ~26% in the control + OGD group and by 27% in the control + H2O2 group. Following the knockdown of Srxn1, cell viability decreased more significantly (sh-Srxn1 + OGD group and sh-Srxn1 + H2O2 group) compared with the control + OGD (or H2O2) group (P<0.05). We also determined astrocyte damage by LDH assay (Fig. 2B). Exposure to OGD (or H2O2) resulted in an increase in the release of LDH (P<0.05), and following exposure to OGD or H2O2, the knockdown of Srxn1 resulted in the release of LDH increasing more significantly compared with the control + OGD (or H2O2) group (P<0.01; Fig. 2B). These results indicate that Srxn1 may protect astrocytes from OGD- or H2O2-induced damage.


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.

Zhou Y, Zhou Y, Yu S, Wu J, Chen Y, Zhao Y - Int. J. Mol. Med. (2015)

Knockdown of sulfiredoxin 1 (Srxn1) decreases cell viability and increases cell damage following exposure to oxygen-glucose deprivation (OGD) or hydrogen peroxide (H2O2). (A) Cell viability was determined by MTS assay. Exposure to OGD or H2O2 decreased cell viability. Following exposure to OGD or H2O2, the knockdown of Srxn1 promoted the reduction in cell viability. Values are the means ± SEM; *P<0.05, **P<0.01 vs. controls (untreated cells); #P<0.05 vs. control + OGD (or H2O2) group. n=6 from 3 independent experiments. (B) Cell damage was assessed by measuring the lactate dehydrogenase (LDH) levels. Exposure to OGD or H2O2 increased cell damage. Following exposure to OGD or H2O2, the knockdown of Srxn1 increased cell damage even more significantly. n=3 from 3 independent experiments. Values are the means ± SEM; **P<0.01 vs. controls (untreated cells); ##P<0.01 vs. control + OGD (or H2O2) group. NC, negative control.
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Related In: Results  -  Collection

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f2-ijmm-36-01-0043: Knockdown of sulfiredoxin 1 (Srxn1) decreases cell viability and increases cell damage following exposure to oxygen-glucose deprivation (OGD) or hydrogen peroxide (H2O2). (A) Cell viability was determined by MTS assay. Exposure to OGD or H2O2 decreased cell viability. Following exposure to OGD or H2O2, the knockdown of Srxn1 promoted the reduction in cell viability. Values are the means ± SEM; *P<0.05, **P<0.01 vs. controls (untreated cells); #P<0.05 vs. control + OGD (or H2O2) group. n=6 from 3 independent experiments. (B) Cell damage was assessed by measuring the lactate dehydrogenase (LDH) levels. Exposure to OGD or H2O2 increased cell damage. Following exposure to OGD or H2O2, the knockdown of Srxn1 increased cell damage even more significantly. n=3 from 3 independent experiments. Values are the means ± SEM; **P<0.01 vs. controls (untreated cells); ##P<0.01 vs. control + OGD (or H2O2) group. NC, negative control.
Mentions: Following exposure to OGD or H2O2, cell viability was determined by MTS assay (Fig. 2A). Compared with the control group, following exposure to OGD or H2O2, cell viability was decreased by ~26% in the control + OGD group and by 27% in the control + H2O2 group. Following the knockdown of Srxn1, cell viability decreased more significantly (sh-Srxn1 + OGD group and sh-Srxn1 + H2O2 group) compared with the control + OGD (or H2O2) group (P<0.05). We also determined astrocyte damage by LDH assay (Fig. 2B). Exposure to OGD (or H2O2) resulted in an increase in the release of LDH (P<0.05), and following exposure to OGD or H2O2, the knockdown of Srxn1 resulted in the release of LDH increasing more significantly compared with the control + OGD (or H2O2) group (P<0.01; Fig. 2B). These results indicate that Srxn1 may protect astrocytes from OGD- or H2O2-induced damage.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Chongqing Medical University, Chongqing 400016, P.R. China.

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.

No MeSH data available.


Related in: MedlinePlus