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Activation of peroxisome proliferator-activated receptor-gamma by glitazones reduces the expression and release of monocyte chemoattractant protein-1 in human mesothelial cells.

Sauter M, Kastenmüller K, Belling F, Wörnle M, Ladurner R, Mussack T, Sitter T - Mediators Inflamm. (2012)

Bottom Line: Activation of this receptor via rosiglitazone (0,1-10 μmol/L) resulted in significantly reduced amounts of mesothelial MCP-1 release as well as MCP-1 mRNA.Rosiglitazone was also effective in reducing TNFα-induced enhanced secretion of MCP-1.Our findings indicate that glitazones are effective in reducing constitutive and TNFα-stimulated mesothelial MCP-1 mRNA expression and release.

View Article: PubMed Central - PubMed

Affiliation: Medizinische Poliklinik-Innenstadt, Klinikum der Universitaet Muenchen, Muenchen, Germany. matthias.sauter@med.uni-muenchen.de

ABSTRACT
Human peritoneal mesothelial cells (MC) play an important role in inflammatory processes of the peritoneal cavity by producing various cytokines and chemokines, such as monocyte chemoattractant protein-1 (MCP-1). The present study was designed to assess the effect of the peroxisome proliferator-activated receptor-gamma- (PPARγ-) activator rosiglitazone on the mesothelial MCP-1 expression and release. Primary cultures of MC were obtained from omental tissue. MCP-1 antigen concentrations were measured in the cell supernatant by ELISA and MCP-1 mRNA levels by real-time polymerase chain reaction. The presence of PPARγ on MC was assayed in a Western Blot analysis. MC constitutively express PPARγ. Activation of this receptor via rosiglitazone (0,1-10 μmol/L) resulted in significantly reduced amounts of mesothelial MCP-1 release as well as MCP-1 mRNA. The use of the PPARγ inhibitor GW-9662 could completely prevent the rosiglitazone effects. Rosiglitazone was also effective in reducing TNFα-induced enhanced secretion of MCP-1. Our findings indicate that glitazones are effective in reducing constitutive and TNFα-stimulated mesothelial MCP-1 mRNA expression and release.

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Western Blot analysis of PPARγ-1 in two different isolates o unstimulated mesothelial cells. Total protein was extracted from unstimulated peritoneal mesothelial cells or human breast carcinoma cells (positive control) and were analysed using the Western Blot technique. The amount of total protein loaded is indicated. Pos. ctrl: positive control; neg. ctrl: negative control.
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fig1: Western Blot analysis of PPARγ-1 in two different isolates o unstimulated mesothelial cells. Total protein was extracted from unstimulated peritoneal mesothelial cells or human breast carcinoma cells (positive control) and were analysed using the Western Blot technique. The amount of total protein loaded is indicated. Pos. ctrl: positive control; neg. ctrl: negative control.

Mentions: We evaluated the presence of PPAR-γ1 on human MC via Western Blot. Analysis of the extracted total protein of unstimulated MC showed a single band at 67 kDa as has been described for PPAR-γ1 and was comparable to the recommended positive control (protein from human breast carcinoma cells) (Figure 1).


Activation of peroxisome proliferator-activated receptor-gamma by glitazones reduces the expression and release of monocyte chemoattractant protein-1 in human mesothelial cells.

Sauter M, Kastenmüller K, Belling F, Wörnle M, Ladurner R, Mussack T, Sitter T - Mediators Inflamm. (2012)

Western Blot analysis of PPARγ-1 in two different isolates o unstimulated mesothelial cells. Total protein was extracted from unstimulated peritoneal mesothelial cells or human breast carcinoma cells (positive control) and were analysed using the Western Blot technique. The amount of total protein loaded is indicated. Pos. ctrl: positive control; neg. ctrl: negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3306974&req=5

fig1: Western Blot analysis of PPARγ-1 in two different isolates o unstimulated mesothelial cells. Total protein was extracted from unstimulated peritoneal mesothelial cells or human breast carcinoma cells (positive control) and were analysed using the Western Blot technique. The amount of total protein loaded is indicated. Pos. ctrl: positive control; neg. ctrl: negative control.
Mentions: We evaluated the presence of PPAR-γ1 on human MC via Western Blot. Analysis of the extracted total protein of unstimulated MC showed a single band at 67 kDa as has been described for PPAR-γ1 and was comparable to the recommended positive control (protein from human breast carcinoma cells) (Figure 1).

Bottom Line: Activation of this receptor via rosiglitazone (0,1-10 μmol/L) resulted in significantly reduced amounts of mesothelial MCP-1 release as well as MCP-1 mRNA.Rosiglitazone was also effective in reducing TNFα-induced enhanced secretion of MCP-1.Our findings indicate that glitazones are effective in reducing constitutive and TNFα-stimulated mesothelial MCP-1 mRNA expression and release.

View Article: PubMed Central - PubMed

Affiliation: Medizinische Poliklinik-Innenstadt, Klinikum der Universitaet Muenchen, Muenchen, Germany. matthias.sauter@med.uni-muenchen.de

ABSTRACT
Human peritoneal mesothelial cells (MC) play an important role in inflammatory processes of the peritoneal cavity by producing various cytokines and chemokines, such as monocyte chemoattractant protein-1 (MCP-1). The present study was designed to assess the effect of the peroxisome proliferator-activated receptor-gamma- (PPARγ-) activator rosiglitazone on the mesothelial MCP-1 expression and release. Primary cultures of MC were obtained from omental tissue. MCP-1 antigen concentrations were measured in the cell supernatant by ELISA and MCP-1 mRNA levels by real-time polymerase chain reaction. The presence of PPARγ on MC was assayed in a Western Blot analysis. MC constitutively express PPARγ. Activation of this receptor via rosiglitazone (0,1-10 μmol/L) resulted in significantly reduced amounts of mesothelial MCP-1 release as well as MCP-1 mRNA. The use of the PPARγ inhibitor GW-9662 could completely prevent the rosiglitazone effects. Rosiglitazone was also effective in reducing TNFα-induced enhanced secretion of MCP-1. Our findings indicate that glitazones are effective in reducing constitutive and TNFα-stimulated mesothelial MCP-1 mRNA expression and release.

Show MeSH
Related in: MedlinePlus