Yonsei Med J.  2015 Mar;56(2):426-432. 10.3349/ymj.2015.56.2.426.

Effects of Interleukin-13 and Montelukast on the Expression of Zonula Occludens-1 in Human Podocytes

• Park SJ
• Saleem MA
• Nam JA
• Ha TS
• Shin JI

• KMID: 2070020
• DOI: http://doi.org/10.3349/ymj.2015.56.2.426

Abstract

PURPOSE
The aim of this study was to investigate whether pathologic changes in zonula occludens-1 (ZO-1) are induced by interleukin-13 (IL-13) in the experimental minimal-change nephrotic syndrome (MCNS) model and to determine whether montelukast, a leukotriene receptor antagonist, has an effect on ZO-1 restoration in cultured human podocytes.
MATERIALS AND METHODS
Human podocytes cultured on bovine serum albumin-coated plates were treated with different doses of IL-13 and montelukast and then examined for distribution using confocal microscopy and for ZO-1 protein levels using Western blotting.
RESULTS
ZO-1 was internalized and shown to accumulate in the cytoplasm of human podocytes in an IL-13 dose-dependent manner. High doses (50 and 100 ng/mL) of IL-13 decreased the levels of ZO-1 protein at 12 and 24 h (both p<0.01; n=3), which were significantly reversed by a high dose (0.5 microM) montelukast treatment (p<0.01; n=3).
CONCLUSION
Our results suggest that IL-13 alters the expression of ZO-1, and such alterations in the content and distribution of ZO-1 may be relevant in the pathogenesis of proteinuria in the MCNS model.

Keyword

Interleukin-13; zonula occludens-1; podocytes; leukotriene receptor antagonists

MeSH Terms

Acetates/*pharmacology
Blotting, Western
Dose-Response Relationship, Drug
Humans
Interleukin-13/*pharmacology
Leukotriene Antagonists/*pharmacology
Microscopy, Confocal
Podocytes/*drug effects/metabolism
Proteinuria/pathology
Quinolines/*pharmacology
Tight Junctions
Zonula Occludens-1 Protein/*metabolism
Acetates
Interleukin-13
Leukotriene Antagonists
Quinolines
Zonula Occludens-1 Protein

Figure

• 

  Fig. 1 Distributional changes in ZO-1 by IL-13 in human podocytes. ZO-1 was distributed at the peripheral cell membrane and colocalized with β-catenin and actin filament at cell-to-cell contact junctions. High concentrations of IL-13 suppressed and disrupted the immunostaining and linearity of ZO-1 proteins, and accumulated ZO-1 proteins into the cytoplasm around nucleus (A), which improved by treatment with 0.5 µM montelukast (B). Magnification: 1000×; Scale bar=20 µm. ZO-1, zonula occludens-1; IL-13, interleukin-13.

• 

  Fig. 2 Effects of IL-13 on ZO-1 protein levels in cultured human podocytes as assayed by Western blotting. ZO-1 levels significantly decreased at IL-13 concentrations of more than 50 ng/mL at 12 and 24 h incubations, compared with the control. Data on the densitometric analysis of the ZO-1/β-tubulin ratio are expressed as mean±SD. Control (100%): the value of (-). *p<0.05. †p<0.01. ZO-1, zonula occludens-1; IL-13, interleukin-13.

• 

  Fig. 3 Effects of montelukast on ZO-1 protein levels in cultured human podocytes assayed by Western blotting. Montelukast (0.5 µM) significantly increased ZO-1 levels in human podocytes treated with IL-13. *p<0.05. †p<0.01. ZO-1, zonula occludens-1; IL-13, interleukin-13.

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