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Nitric oxide mediates prolyl hydroxylase 3 expression in mesangial cells and in glomerulonephritis

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Abstract

Renal mesangial cells are regarded as main players in glomerular inflammatory diseases. To investigate a possible crosstalk between inflammatory and hypoxia-driven signaling processes, we stimulated cultured mouse mesangial cells with different inflammatory agents and analyzed the expression of prolyl hydroxylase domain containing proteins (PHDs), the main regulators of hypoxia-inducible factor (HIF) stability. Administration of IL-1β (1 nM) and TNF-α (1 nM), a combination further referred to as cytokine mix (CM), resulted in a fivefold increase in PHD3 but not PHD1 and PHD2 mRNA expression compared to untreated controls. In contrast, a combination of IL-1β, TNF-α with lipopolysaccharide (10 μg/ml), and interferon-γ (20 ng/ml) designated as CM+ showed a high (60-fold) induction of PHD3 and a moderate (twofold) induction of PHD2 mRNA expression. Interestingly, CM+ but not CM induced the expression of inducible NO synthase and endogenously produced NO was responsible for the immense induction of PHD3 in mesangial cells treated with CM+. We found that CM+ affected PHD3 expression mainly via the NO/HIF axis, whereas PHD3 regulation by CM occurred in a NF-κB-dependent manner. In turn, silencing of PHD3 expression resulted in a decrease in the mRNA expression of ICAM-1, MIP-2, MCP-1, and CXCL-10, which are under control of NF-κB. In a rat model of mesangio-proliferative glomerulonephritis, PHD3 mRNA and protein expression was markedly induced and this effect was nearly abolished when rats were treated with the iNOS-specific inhibitor L-NIL, thus confirming our findings also in vivo.

Key message

  • PHD3 expression induced by cytokines is NF-κB dependent in mesangial cells.

  • Endogenously produced NO further augments PHD3 expression via HIF-1α.

  • PHD3 expression is induced by NO in anti-Thy-1 glomerulonephritis.

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Acknowledgements

The authors thank Ute Schmidt and Riad Haceni for valuable technical support and Dr. Miriam Pleskova for performing experiments with rat mesangial cells. The work was supported by the German Research Foundation (SFB 815, project A7 for JP, project A5 for LS; SFB 1039, project B2; SFB 1177, project C2; SCHA 1082/6-1 to LS, and LOEWE program Ub-Net to LS). AA and MH were supported by grants of the Ministry of Higher Education of the Arab Republic of Egypt.

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Authors and Affiliations

  1. Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany

    Ahmed Aglan, Sebastian Longen, Yvette Köhler, Mohamed Hassan, Martina Beck, Claudia Tredup, Meike Boosen, Tzung-Harn Louise Hsieh, Liliana Schaefer, Karl-Friedrich Beck & Josef Pfeilschifter

  2. Department of Biochemistry, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt

    Ahmed Aglan

  3. Institut für Biochemie, Pathobiochemie, Goethe Universität, Universitätsklinikum Frankfurt, Frankfurt am Main, Germany

    Nathalie Dehne

  4. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Al-Azhar University, Assiut, Egypt

    Mohamed Hassan

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  1. Ahmed Aglan
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Correspondence to Karl-Friedrich Beck.

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Aglan, A., Longen, S., Dehne, N. et al. Nitric oxide mediates prolyl hydroxylase 3 expression in mesangial cells and in glomerulonephritis. J Mol Med 95, 257–271 (2017). https://doi.org/10.1007/s00109-016-1503-3

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  • DOI: https://doi.org/10.1007/s00109-016-1503-3

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