title: Nitric oxide mediates prolyl hydroxylase 3 expression in mesangial cells and in glomerulonephritis.
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.
  
  
  
  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.
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.|Department of Biochemistry, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Aglan
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Longen
authors.affiliation: Institut für Biochemie, Pathobiochemie, Goethe Universität, Universitätsklinikum Frankfurt, Frankfurt am Main, Germany.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Dehne
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.
authors.forename: Yvette
authors.initials: Y
authors.lastname: Köhler
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.|Department of Pharmacology and Toxicology, Faculty of Pharmacy, Al-Azhar University, Assiut, Egypt.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Hassan
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Beck
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Tredup
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.
authors.forename: Meike
authors.initials: M
authors.lastname: Boosen
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.
authors.forename: Tzung-Harn Louise
authors.initials: TL
authors.lastname: Hsieh
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.
authors.forename: Liliana
authors.initials: L
authors.lastname: Schaefer
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany. k.f.beck@em.uni-frankfurt.de.
authors.forename: Karl-Friedrich
authors.initials: KF
authors.lastname: Beck
authors.affiliation: Pharmazentrum Frankfurt/ZAFES, Goethe Universität, Universitätsklinikum Frankfurt, Theodor-Stern-Kai 7, D 60590, Frankfurt am Main, Germany.
authors.forename: Josef
authors.initials: J
authors.lastname: Pfeilschifter
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005921:Glomerulonephritis / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D006801:Humans; D053583:Interleukin-1beta / Q000276:immunology; D050527:Mesangial Cells / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000276:immunology; D009569:Nitric Oxide / Q000276:immunology*; D011393:Procollagen-Proline Dioxygenase / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology; D015854:Up-Regulation
chemical_list: D053583:Interleukin-1beta; D016328:NF-kappa B; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D011393:Procollagen-Proline Dioxygenase; C030606:proline, 2-oxoglutarate 3-dioxygenase
keywords: Mesangial cells; Nitric oxide; Prolyl hydroxylase domain containing protein 3