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Int. J. Mol. Sci. 2014, 15(4), 6592-6608; doi:10.3390/ijms15046592

A20 Overexpression Inhibits Lipopolysaccharide-Induced NF-κB Activation, TRAF6 and CD40 Expression in Rat Peritoneal Mesothelial Cells

1,†,* , 2,†
,
1
,
1
and
1
1
Department of Nephrology, the Affiliated Hospital, Hangzhou Normal University, Hangzhou 310015, Zhejiang, China
2
Division of Cardiology, Hangzhou Red Cross Hospital, Hangzhou 310003, Zhejiang, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 11 February 2014 / Revised: 4 March 2014 / Accepted: 24 March 2014 / Published: 17 April 2014
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

Zinc finger protein A20 is a key negative regulator of inflammation. However, whether A20 may affect inflammation during peritoneal dialysis (PD)-associated peritonitis is still unclear. This study was aimed to investigate the effect of A20 overexpression on lipopolysaccharide (LPS)-induced inflammatory response in rat peritoneal mesothelial cells (RPMCs). Isolated and cultured RPMCs in vitro. Plasmid pGEM-T easy-A20 was transfected into RPMCs by Lipofectamine™2000. The protein expression of A20, phospho-IκBα, IκBα, TNF receptor-associated factor (TRAF) 6 and CD40 were analyzed by Western blot. The mRNA expression of TRAF6, CD40, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were determined by real time-PCR. NF-κB p65 DNA binding activity, IL-6 and TNF-α levels in cells culture supernatant were determined by ELISA. Our results revealed that RPMCs overexpression of A20 lead to significant decrease of LPS-induced IκBα phosphorylation and NF-κB DNA binding activity (all p < 0.01). In addition, A20 also attenuated the expression of TRAF6, CD40, IL-6 and TNF-α as well as levels of IL-6 and TNF-α in cells culture supernatant (all p < 0.05). However, A20 only partly inhibited CD40 expression. Our study indicated that A20 overexpression may depress the inflammatory response induced by LPS in cultured RPMCs through negatively regulated the relevant function of adaptors in LPS signaling pathway. View Full-Text
Keywords: rat peritoneal mesothelial cells; Zinc finger protein A20; NF-κB; TRAF6; CD40 rat peritoneal mesothelial cells; Zinc finger protein A20; NF-κB; TRAF6; CD40
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This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Zou, X.-L.; Pei, D.-A.; Yan, J.-Z.; Xu, G.; Wu, P. A20 Overexpression Inhibits Lipopolysaccharide-Induced NF-κB Activation, TRAF6 and CD40 Expression in Rat Peritoneal Mesothelial Cells. Int. J. Mol. Sci. 2014, 15, 6592-6608.

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