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




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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.



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





Autor: Xun-Liang Zou 1,†,* , De-An Pei 2,†, Ju-Zhen Yan 1, Gang Xu 1 and Ping Wu 1

Fuente: http://mdpi.com/



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