Paricalcitol protects against TGF-β1-induced fibrotic responses in hypoxia and stabilises HIF-α in renal epithelia

Exp Cell Res. 2015 Jan 15;330(2):371-381. doi: 10.1016/j.yexcr.2014.07.034. Epub 2014 Aug 5.

Abstract

Epithelial injury and tubulointerstitial fibrosis (TIF) within a hypoxic microenvironment are associated with progressive loss of renal function in chronic kidney disease [CKD]. Transforming growth factor beta-1 (TGF-β1) is an important mediator of renal fibrosis. Growing evidence suggests that Vitamin D [1,25-(OH)2D] and its analogues may have a renoprotective effect in CKD. Here we examined the protective effect of the vitamin D analogue paricalcitol [PC; 19-nor-1α,3β,25-trihydroxy-9,10-secoergosta-5(Z),7(E) 22(E)-triene] on the responses of human renal epithelial cells to TGF-β1. PC attenuated TGF-β1-induced Smad 2 phosphorylation and upregulation of the Notch ligand Jagged-1, α-smooth muscle actin and thrombospondin-1 and prevented the TGF-β1-mediated loss of E-Cadherin. To mimic the hypoxic milieu of CKD we cultured renal epithelial cells in hypoxia [1% O2] and observed similar attenuation by PC of TGF-β1-induced fibrotic responses. Furthermore, in cells cultured in normoxia [21% O2], PC induced an accumulation of hypoxia-inducible transcription factors (HIF) 1α and HIF-2α in a time and concentration [1 µM-2 µM] dependent manner. Here, PC-induced HIF stabilisation was dependent on activation of the PI-3Kinase pathway. This is the first study to demonstrate regulation of the HIF pathway by PC which may have importance in the mechanism underlying renoprotection by PC.

Keywords: Fibrosis; HIF; Hypoxia; Kidney; Paricalcitol; TGF-β1; Vitamin D.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / biosynthesis
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cadherins / metabolism
  • Calcium-Binding Proteins / biosynthesis
  • Cell Hypoxia
  • Cell Line, Transformed
  • Epithelial Cells / drug effects*
  • Epithelial Cells / pathology
  • Ergocalciferols / pharmacology*
  • Fibrosis
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
  • Intercellular Signaling Peptides and Proteins / biosynthesis
  • Jagged-1 Protein
  • Kidney / drug effects*
  • Kidney / pathology*
  • Membrane Proteins / biosynthesis
  • Nephritis, Interstitial / pathology
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphorylation
  • Protein Stability
  • RNA Interference
  • Serrate-Jagged Proteins
  • Smad2 Protein / metabolism
  • Thrombospondin 1 / biosynthesis
  • Transforming Growth Factor beta1 / antagonists & inhibitors*
  • Transforming Growth Factor beta1 / metabolism

Substances

  • ACTA2 protein, human
  • Actins
  • Basic Helix-Loop-Helix Transcription Factors
  • Cadherins
  • Calcium-Binding Proteins
  • Ergocalciferols
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Intercellular Signaling Peptides and Proteins
  • JAG1 protein, human
  • Jagged-1 Protein
  • Membrane Proteins
  • Serrate-Jagged Proteins
  • Smad2 Protein
  • Thrombospondin 1
  • Transforming Growth Factor beta1
  • endothelial PAS domain-containing protein 1
  • 1,25-dihydroxyergocalciferol
  • paricalcitol
  • Phosphatidylinositol 3-Kinases