15-Deoxy-Δ(12,14)-prostaglandin J(2) modulates lipopolysaccharide-induced chemokine expression by blocking nuclear factor-κB activation via peroxisome proliferator activated receptor-γ-independent mechanism in renal tubular epithelial cells

Nephron Exp Nephrol. 2013;123(1-2):1-10. doi: 10.1159/000353232. Epub 2013 Jul 24.

Abstract

Background/aims: Inflammation is an unavoidable milieu for renal tubular cells during the development of renal tubulointerstitial fibrosis. It has been demonstrated that chemokines including monocyte chemoattractant protein-1 (MCP-1) and IL-8 are related to tubulointerstitial lesions. 15d-PGJ2 may modulate renal tubulointerstitial fibrosis progression via anti-inflammatory effects. However, no information is known about the effects of 15d-PGJ2 on chemokine expression in human proximal renal tubular cells (HPTECs) under inflammation.

Methods: In the present study, HPTECs (HK-2 cells) were stimulated with lipopolysaccharide (LPS) only, or preincubated with 15d-PGJ2. IL-8 and MCP-1 expressions were determined by real-time PCR and ELISA. Nuclear factor-κB (NF-κB) location was detected by immunofluorescence analysis. The p-IKK, p-IκBα and p65/p50 were analyzed by immunoblotting. To investigate the mechanism of inhibitory effects of 15d-PGJ2, the PPAR-γ gene was effectively silenced in HK-2 cells using specific siRNA.

Results: The results showed that application of LPS significantly increased IL-8 and MCP-1 production. Phosphorylation of IκBα, IKK and nucleus translocation of NF-κB significantly increased in LPS-stimulated HK-2 cells. 15d-PGJ2 downregulated LPS-induced IL-8 and MCP-1 production. Interestingly, in PPAR-γ-deficient HK-2 cells, 15d-PGJ2 was still capable of inhibiting chemokines expression and attenuating phosphorylation of IκBα and nucleus translocation of NF-κB.

Conclusion: Collectively, these results suggest that 15d-PGJ2 exerts anti-inflammatory actions on HK-2 cells by attenuating chemokines expression. 15d-PGJ2 inhibits chemokines expression via a PPAR-γ-independent way, which is related to block NF-κB pathway. Since NF-κB is an important regulator of the response of HPTECs to injury, PPAR-γ agonists may represent a key pharmacological target for ameliorating inflammation-associated tubulointerstitial fibrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus / drug effects
  • Cell Line
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism
  • Chemokine CCL2 / genetics
  • Chemokine CCL2 / metabolism
  • Chemokines / genetics
  • Chemokines / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism
  • Gene Expression / drug effects
  • Humans
  • I-kappa B Proteins / metabolism
  • Immunoblotting
  • Interleukin-8 / genetics
  • Interleukin-8 / metabolism
  • Kidney Tubules, Proximal / cytology
  • Lipopolysaccharides / pharmacology*
  • Microscopy, Fluorescence
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / metabolism*
  • PPAR gamma / genetics
  • PPAR gamma / metabolism
  • Phosphorylation / drug effects
  • Prostaglandin D2 / analogs & derivatives*
  • Prostaglandin D2 / pharmacology
  • RNA Interference
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / drug effects

Substances

  • 15-deoxy-delta(12,14)-prostaglandin J2
  • Chemokine CCL2
  • Chemokines
  • I-kappa B Proteins
  • Interleukin-8
  • Lipopolysaccharides
  • NF-kappa B
  • NFKBIA protein, human
  • PPAR gamma
  • NF-KappaB Inhibitor alpha
  • Prostaglandin D2