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Glucocorticoids Induce Cardiac Fibrosis via Mineralocorticoid Receptor in Oxidative Stress: Contribution of Elongation Factor Eleven-Nineteen Lysine-Rich Leukemia (ELL).

Omori Y, Mano T, Ohtani T, Sakata Y, Takeda Y, Tamaki S, Tsukamoto Y, Miwa T, Yamamoto K, Komuro I - Yonago Acta Med (2014)

Bottom Line: The MR antagonist eplerenone attenuated corticosterone-induced collagen synthesis assessed by [(3)H]proline incorporation in rat neonatal cultured cardiac fibroblasts in the presence of H2O2, as an oxidative stress but not in the absence of H2O2.H2O2 increased the ELL expression levels and MR-bound ELL.Eplerenone did not attenuate corticosterone-induced increase of [(3)H]proline incorporation in the presence of H2O2 after knockdown of ELL expression using small interfering RNA in cardiac fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine, Suita 565-0871, Japan.

ABSTRACT

Background: Cardiac fibrosis is considered to be a crucial factor in the development of heart failure. Blockade of the mineralocorticoid receptor (MR) attenuated cardiac fibrosis and improved the prognosis of patients with chronic heart failure but the ligand for MR and the regulatory mechanism of MR pathway in the diseased heart are unclear. Here, we investigated whether glucocorticoids can promote cardiac fibrosis through MR in oxidative stress and the involvement of elongation factor eleven-nineteen lysine-rich leukemia (ELL), a co-activator of MR, in this pathway.

Methods and results: The MR antagonist eplerenone attenuated corticosterone-induced collagen synthesis assessed by [(3)H]proline incorporation in rat neonatal cultured cardiac fibroblasts in the presence of H2O2, as an oxidative stress but not in the absence of H2O2. H2O2 increased the ELL expression levels and MR-bound ELL. ELL expression levels and MR-bound ELL were also increased in the left ventricle of heart failure model rats with significant fibrosis and enhanced oxidative stress. Eplerenone did not attenuate corticosterone-induced increase of [(3)H]proline incorporation in the presence of H2O2 after knockdown of ELL expression using small interfering RNA in cardiac fibroblasts.

Conclusion: Glucocorticoids can promote cardiac fibrosis via MR in oxidative stress, and oxidative stress modulates MR response to glucocorticoids through the interaction with ELL. Preventing cardiac fibrosis by modulating glucocorticoid-MR-ELL pathway may become a new therapeutic strategy for heart failure.

No MeSH data available.


Related in: MedlinePlus

The effect of H2O2 on the expression of ELL, MR and 11betaHSD2, and ELL-MRinteraction in cultured cardiac fibroblasts. Cardiac fibroblasts were treated withH2O2 (1 μmol/L) for 48 h. Cardiac fibroblasts without H2O2were served as Ctl.A: RT-qPCR was performed to evaluate the mRNA levels of ELL, MR and 11betaHSD2in the cardiac fibroblasts (n = 5 for each group).B: WB analysis was performed to evaluate the protein levels of ELL and MR in thecardiac fibroblasts. The images (upper) and the results of densitometry (lower) of WB analysis(n = 4 for each group).Ctl was arbitrarily set to 1. Data are expressed as mean values ± SD.*P < 0.05 versus Ctl.C: Cell lysate of cardiac fibroblasts was subjected to IP with anti-MR antibody.Immunoprecipitates were subsequently analyzed by Western blotting with anti-ELL antibody.Ctl, control; ELL, elongation factor eleven-nineteen lysine-rich leukemia; IP, immunoprecipitation;RT-qPCR, real-time quantitative reverse transcriptase-PCR; MR, mineralocorticoid receptor;WB, Western blot.
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fig_002: The effect of H2O2 on the expression of ELL, MR and 11betaHSD2, and ELL-MRinteraction in cultured cardiac fibroblasts. Cardiac fibroblasts were treated withH2O2 (1 μmol/L) for 48 h. Cardiac fibroblasts without H2O2were served as Ctl.A: RT-qPCR was performed to evaluate the mRNA levels of ELL, MR and 11betaHSD2in the cardiac fibroblasts (n = 5 for each group).B: WB analysis was performed to evaluate the protein levels of ELL and MR in thecardiac fibroblasts. The images (upper) and the results of densitometry (lower) of WB analysis(n = 4 for each group).Ctl was arbitrarily set to 1. Data are expressed as mean values ± SD.*P < 0.05 versus Ctl.C: Cell lysate of cardiac fibroblasts was subjected to IP with anti-MR antibody.Immunoprecipitates were subsequently analyzed by Western blotting with anti-ELL antibody.Ctl, control; ELL, elongation factor eleven-nineteen lysine-rich leukemia; IP, immunoprecipitation;RT-qPCR, real-time quantitative reverse transcriptase-PCR; MR, mineralocorticoid receptor;WB, Western blot.

Mentions: The mRNA and the protein levels of ELL were increased in the cultured cardiac fibroblasts underoxidative stress with H2O2 (Figs. 2A andB). IP assay revealed the presence of ELL-MR binding and that the amount of ELL bound to MR wasincreased in the cardiac fibroblasts incubated in the presence of H2O2(Fig. 2C).


Glucocorticoids Induce Cardiac Fibrosis via Mineralocorticoid Receptor in Oxidative Stress: Contribution of Elongation Factor Eleven-Nineteen Lysine-Rich Leukemia (ELL).

Omori Y, Mano T, Ohtani T, Sakata Y, Takeda Y, Tamaki S, Tsukamoto Y, Miwa T, Yamamoto K, Komuro I - Yonago Acta Med (2014)

The effect of H2O2 on the expression of ELL, MR and 11betaHSD2, and ELL-MRinteraction in cultured cardiac fibroblasts. Cardiac fibroblasts were treated withH2O2 (1 μmol/L) for 48 h. Cardiac fibroblasts without H2O2were served as Ctl.A: RT-qPCR was performed to evaluate the mRNA levels of ELL, MR and 11betaHSD2in the cardiac fibroblasts (n = 5 for each group).B: WB analysis was performed to evaluate the protein levels of ELL and MR in thecardiac fibroblasts. The images (upper) and the results of densitometry (lower) of WB analysis(n = 4 for each group).Ctl was arbitrarily set to 1. Data are expressed as mean values ± SD.*P < 0.05 versus Ctl.C: Cell lysate of cardiac fibroblasts was subjected to IP with anti-MR antibody.Immunoprecipitates were subsequently analyzed by Western blotting with anti-ELL antibody.Ctl, control; ELL, elongation factor eleven-nineteen lysine-rich leukemia; IP, immunoprecipitation;RT-qPCR, real-time quantitative reverse transcriptase-PCR; MR, mineralocorticoid receptor;WB, Western blot.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4209341&req=5

fig_002: The effect of H2O2 on the expression of ELL, MR and 11betaHSD2, and ELL-MRinteraction in cultured cardiac fibroblasts. Cardiac fibroblasts were treated withH2O2 (1 μmol/L) for 48 h. Cardiac fibroblasts without H2O2were served as Ctl.A: RT-qPCR was performed to evaluate the mRNA levels of ELL, MR and 11betaHSD2in the cardiac fibroblasts (n = 5 for each group).B: WB analysis was performed to evaluate the protein levels of ELL and MR in thecardiac fibroblasts. The images (upper) and the results of densitometry (lower) of WB analysis(n = 4 for each group).Ctl was arbitrarily set to 1. Data are expressed as mean values ± SD.*P < 0.05 versus Ctl.C: Cell lysate of cardiac fibroblasts was subjected to IP with anti-MR antibody.Immunoprecipitates were subsequently analyzed by Western blotting with anti-ELL antibody.Ctl, control; ELL, elongation factor eleven-nineteen lysine-rich leukemia; IP, immunoprecipitation;RT-qPCR, real-time quantitative reverse transcriptase-PCR; MR, mineralocorticoid receptor;WB, Western blot.
Mentions: The mRNA and the protein levels of ELL were increased in the cultured cardiac fibroblasts underoxidative stress with H2O2 (Figs. 2A andB). IP assay revealed the presence of ELL-MR binding and that the amount of ELL bound to MR wasincreased in the cardiac fibroblasts incubated in the presence of H2O2(Fig. 2C).

Bottom Line: The MR antagonist eplerenone attenuated corticosterone-induced collagen synthesis assessed by [(3)H]proline incorporation in rat neonatal cultured cardiac fibroblasts in the presence of H2O2, as an oxidative stress but not in the absence of H2O2.H2O2 increased the ELL expression levels and MR-bound ELL.Eplerenone did not attenuate corticosterone-induced increase of [(3)H]proline incorporation in the presence of H2O2 after knockdown of ELL expression using small interfering RNA in cardiac fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine, Suita 565-0871, Japan.

ABSTRACT

Background: Cardiac fibrosis is considered to be a crucial factor in the development of heart failure. Blockade of the mineralocorticoid receptor (MR) attenuated cardiac fibrosis and improved the prognosis of patients with chronic heart failure but the ligand for MR and the regulatory mechanism of MR pathway in the diseased heart are unclear. Here, we investigated whether glucocorticoids can promote cardiac fibrosis through MR in oxidative stress and the involvement of elongation factor eleven-nineteen lysine-rich leukemia (ELL), a co-activator of MR, in this pathway.

Methods and results: The MR antagonist eplerenone attenuated corticosterone-induced collagen synthesis assessed by [(3)H]proline incorporation in rat neonatal cultured cardiac fibroblasts in the presence of H2O2, as an oxidative stress but not in the absence of H2O2. H2O2 increased the ELL expression levels and MR-bound ELL. ELL expression levels and MR-bound ELL were also increased in the left ventricle of heart failure model rats with significant fibrosis and enhanced oxidative stress. Eplerenone did not attenuate corticosterone-induced increase of [(3)H]proline incorporation in the presence of H2O2 after knockdown of ELL expression using small interfering RNA in cardiac fibroblasts.

Conclusion: Glucocorticoids can promote cardiac fibrosis via MR in oxidative stress, and oxidative stress modulates MR response to glucocorticoids through the interaction with ELL. Preventing cardiac fibrosis by modulating glucocorticoid-MR-ELL pathway may become a new therapeutic strategy for heart failure.

No MeSH data available.


Related in: MedlinePlus