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Epoetin delta reduces oxidative stress in primary human renal tubular cells.

De Beuf A, Hou XH, D'Haese PC, Verhulst A - J. Biomed. Biotechnol. (2010)

Bottom Line: Erythropoietin (EPO) exerts (renal) tissue protective effects.Oxidative stress reduction went along with the upregulation of renoprotective genes.Whilst three of these, heme oxygenase-1 (HO-1), aquaporin-1 (AQP-1), and B-cell CLL/lymphoma 2 (Bcl-2) have already been associated with EPO-induced renoprotection, this study for the first time suggests carboxypeptidase M (CPM), dipeptidyl peptidase IV (DPPIV), and cytoglobin (Cygb) to play a role in this process.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Pathophysiology, Faculties of Medicine and Biomedical, Pharmaceutical and Veterinary Sciences, University of Antwerp, Universiteitsplein 1, 2610 Antwerp, Belgium.

ABSTRACT
Erythropoietin (EPO) exerts (renal) tissue protective effects. Since it is unclear whether this is a direct effect of EPO on the kidney or not, we investigated whether EPO is able to protect human renal tubular epithelial cells (hTECs) from oxidative stress and if so which pathways are involved. EPO (epoetin delta) could protect hTECs against oxidative stress by a dose-dependent inhibition of reactive oxygen species formation. This protective effect is possibly related to the membranous expression of the EPO receptor (EPOR) since our data point to the membranous EPOR expression as a prerequisite for this protective effect. Oxidative stress reduction went along with the upregulation of renoprotective genes. Whilst three of these, heme oxygenase-1 (HO-1), aquaporin-1 (AQP-1), and B-cell CLL/lymphoma 2 (Bcl-2) have already been associated with EPO-induced renoprotection, this study for the first time suggests carboxypeptidase M (CPM), dipeptidyl peptidase IV (DPPIV), and cytoglobin (Cygb) to play a role in this process.

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Quantitative real-time RT-PCR analysis of HO-1 (a), AQP-1 (b), and Bcl-2 (c) expressions in mixed hTECs under basal conditions and after GO-induced oxidative stress (1 IU/mL) either or not in the presence of epoetin delta (100 IU/mL). GAPDH was used as endogenous control housekeeping gene. Preconditioning the cells with EPO resulted in a significant upregulation of HO-1 and AQP-1 mRNA. GO-induced oxidative stress further increased HO-1, AQP-1, and Bcl-2 mRNA expressions with maximum levels 60 minutes after induction of oxidative stress. Data are presented as the mean ± SD of triplicate determinations of 2 runs (i.e., 6 values each). *P < .05 versus no GO, °P < .05 versus epoetin delta 0 IU/mL, #P < .05 versus 30 minutes GO incubation.
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fig5: Quantitative real-time RT-PCR analysis of HO-1 (a), AQP-1 (b), and Bcl-2 (c) expressions in mixed hTECs under basal conditions and after GO-induced oxidative stress (1 IU/mL) either or not in the presence of epoetin delta (100 IU/mL). GAPDH was used as endogenous control housekeeping gene. Preconditioning the cells with EPO resulted in a significant upregulation of HO-1 and AQP-1 mRNA. GO-induced oxidative stress further increased HO-1, AQP-1, and Bcl-2 mRNA expressions with maximum levels 60 minutes after induction of oxidative stress. Data are presented as the mean ± SD of triplicate determinations of 2 runs (i.e., 6 values each). *P < .05 versus no GO, °P < .05 versus epoetin delta 0 IU/mL, #P < .05 versus 30 minutes GO incubation.

Mentions: In the present study, the mRNA expression of HO-1, AQP-1, and Bcl-2 was assessed in cultures that were either preincubated with epoetin delta before the induction of oxidative stress or not. Under basal conditions (no GO incubation), preconditioning of hTECs with epoetin delta (100 IU/mL) resulted in a significant upregulation of HO-1 (1.00 ± 0.31 versus 1.56 ± 0.37, P < .05) and AQP-1 (1.00 ± 0.03 versus 2.26 ± 0.12, P < .05) mRNA expression. During GO-induced oxidative stress (1 IU/mL), cultures which were preincubated with epoetin delta (100 IU/mL) showed a further increase in mRNA expression of all three genes investigated as compared to cultures which were not preincubated with epoetin delta, reaching maximum values 60 minutes after the induction of oxidative stress (HO-1: 1.07 ± 0.20 versus 1.48 ± 0.19; AQP-1: 1.61 ± 0.11 versus 3.18 ± 0.22; Bcl-2: 0.97 ± 0.18 versus 1.58 ± 0.40; P < .05) (Figure 5). Interestingly, in cultures in which no epoetin delta-protective effect against oxidative stress was seen also no epoetin delta-induced upregulation of the genes under study could be observed (data not shown).


Epoetin delta reduces oxidative stress in primary human renal tubular cells.

De Beuf A, Hou XH, D'Haese PC, Verhulst A - J. Biomed. Biotechnol. (2010)

Quantitative real-time RT-PCR analysis of HO-1 (a), AQP-1 (b), and Bcl-2 (c) expressions in mixed hTECs under basal conditions and after GO-induced oxidative stress (1 IU/mL) either or not in the presence of epoetin delta (100 IU/mL). GAPDH was used as endogenous control housekeeping gene. Preconditioning the cells with EPO resulted in a significant upregulation of HO-1 and AQP-1 mRNA. GO-induced oxidative stress further increased HO-1, AQP-1, and Bcl-2 mRNA expressions with maximum levels 60 minutes after induction of oxidative stress. Data are presented as the mean ± SD of triplicate determinations of 2 runs (i.e., 6 values each). *P < .05 versus no GO, °P < .05 versus epoetin delta 0 IU/mL, #P < .05 versus 30 minutes GO incubation.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2864893&req=5

fig5: Quantitative real-time RT-PCR analysis of HO-1 (a), AQP-1 (b), and Bcl-2 (c) expressions in mixed hTECs under basal conditions and after GO-induced oxidative stress (1 IU/mL) either or not in the presence of epoetin delta (100 IU/mL). GAPDH was used as endogenous control housekeeping gene. Preconditioning the cells with EPO resulted in a significant upregulation of HO-1 and AQP-1 mRNA. GO-induced oxidative stress further increased HO-1, AQP-1, and Bcl-2 mRNA expressions with maximum levels 60 minutes after induction of oxidative stress. Data are presented as the mean ± SD of triplicate determinations of 2 runs (i.e., 6 values each). *P < .05 versus no GO, °P < .05 versus epoetin delta 0 IU/mL, #P < .05 versus 30 minutes GO incubation.
Mentions: In the present study, the mRNA expression of HO-1, AQP-1, and Bcl-2 was assessed in cultures that were either preincubated with epoetin delta before the induction of oxidative stress or not. Under basal conditions (no GO incubation), preconditioning of hTECs with epoetin delta (100 IU/mL) resulted in a significant upregulation of HO-1 (1.00 ± 0.31 versus 1.56 ± 0.37, P < .05) and AQP-1 (1.00 ± 0.03 versus 2.26 ± 0.12, P < .05) mRNA expression. During GO-induced oxidative stress (1 IU/mL), cultures which were preincubated with epoetin delta (100 IU/mL) showed a further increase in mRNA expression of all three genes investigated as compared to cultures which were not preincubated with epoetin delta, reaching maximum values 60 minutes after the induction of oxidative stress (HO-1: 1.07 ± 0.20 versus 1.48 ± 0.19; AQP-1: 1.61 ± 0.11 versus 3.18 ± 0.22; Bcl-2: 0.97 ± 0.18 versus 1.58 ± 0.40; P < .05) (Figure 5). Interestingly, in cultures in which no epoetin delta-protective effect against oxidative stress was seen also no epoetin delta-induced upregulation of the genes under study could be observed (data not shown).

Bottom Line: Erythropoietin (EPO) exerts (renal) tissue protective effects.Oxidative stress reduction went along with the upregulation of renoprotective genes.Whilst three of these, heme oxygenase-1 (HO-1), aquaporin-1 (AQP-1), and B-cell CLL/lymphoma 2 (Bcl-2) have already been associated with EPO-induced renoprotection, this study for the first time suggests carboxypeptidase M (CPM), dipeptidyl peptidase IV (DPPIV), and cytoglobin (Cygb) to play a role in this process.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Pathophysiology, Faculties of Medicine and Biomedical, Pharmaceutical and Veterinary Sciences, University of Antwerp, Universiteitsplein 1, 2610 Antwerp, Belgium.

ABSTRACT
Erythropoietin (EPO) exerts (renal) tissue protective effects. Since it is unclear whether this is a direct effect of EPO on the kidney or not, we investigated whether EPO is able to protect human renal tubular epithelial cells (hTECs) from oxidative stress and if so which pathways are involved. EPO (epoetin delta) could protect hTECs against oxidative stress by a dose-dependent inhibition of reactive oxygen species formation. This protective effect is possibly related to the membranous expression of the EPO receptor (EPOR) since our data point to the membranous EPOR expression as a prerequisite for this protective effect. Oxidative stress reduction went along with the upregulation of renoprotective genes. Whilst three of these, heme oxygenase-1 (HO-1), aquaporin-1 (AQP-1), and B-cell CLL/lymphoma 2 (Bcl-2) have already been associated with EPO-induced renoprotection, this study for the first time suggests carboxypeptidase M (CPM), dipeptidyl peptidase IV (DPPIV), and cytoglobin (Cygb) to play a role in this process.

Show MeSH
Related in: MedlinePlus