{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,17]],"date-time":"2026-04-17T10:50:46Z","timestamp":1776423046785,"version":"3.51.2"},"reference-count":41,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2014,4,17]],"date-time":"2014-04-17T00:00:00Z","timestamp":1397692800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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\u21222000. The protein expression of A20, phospho-I\u03baB\u03b1, I\u03baB\u03b1, 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-\u03b1 (TNF-\u03b1) were determined by real time-PCR. NF-\u03baB p65 DNA binding activity, IL-6 and TNF-\u03b1 levels in cells culture supernatant were determined by ELISA.  Our results revealed that RPMCs overexpression of A20 lead to significant decrease of LPS-induced I\u03baB\u03b1 phosphorylation and NF-\u03baB DNA binding activity (all p &lt; 0.01). In addition, A20 also attenuated the expression of TRAF6, CD40, IL-6 and TNF-\u03b1 as well as levels of IL-6 and TNF-\u03b1 in cells culture supernatant (all p &lt; 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.<\/jats:p>","DOI":"10.3390\/ijms15046592","type":"journal-article","created":{"date-parts":[[2014,4,17]],"date-time":"2014-04-17T12:21:27Z","timestamp":1397737287000},"page":"6592-6608","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["A20 Overexpression Inhibits Lipopolysaccharide-Induced NF-\u03baB Activation, TRAF6 and CD40 Expression in Rat Peritoneal Mesothelial Cells"],"prefix":"10.3390","volume":"15","author":[{"given":"Xun-Liang","family":"Zou","sequence":"first","affiliation":[{"name":"Department of Nephrology, the Affiliated Hospital, Hangzhou Normal University,  Hangzhou 310015, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"De-An","family":"Pei","sequence":"additional","affiliation":[{"name":"Division of Cardiology, Hangzhou Red Cross Hospital, Hangzhou 310003, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ju-Zhen","family":"Yan","sequence":"additional","affiliation":[{"name":"Department of Nephrology, the Affiliated Hospital, Hangzhou Normal University,  Hangzhou 310015, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gang","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Nephrology, the Affiliated Hospital, Hangzhou Normal University,  Hangzhou 310015, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ping","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Nephrology, the Affiliated Hospital, Hangzhou Normal University,  Hangzhou 310015, Zhejiang, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2014,4,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S35","DOI":"10.1177\/089686080702702s06","article-title":"30 years of peritoneal dialysis development: The past and the future","volume":"27","author":"Krediet","year":"2007","journal-title":"Perit. Dial. Int"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"S104","DOI":"10.1681\/ASN.V13suppl_1s104","article-title":"Peritoneal dialysis in the 21st century: An analysis of current problems and future developments","volume":"13","author":"Gokal","year":"2002","journal-title":"J. Am. Soc. Nephrol"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"S267","DOI":"10.1177\/089686080702702s46","article-title":"Gram-negative peritonitis\u2014The Achilles heel of peritoneal dialysis?","volume":"27","author":"Szeto","year":"2007","journal-title":"Perit. Dial. Int"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.1681\/ASN.2009070694","article-title":"The pathophysiology of the peritoneal membrane","volume":"21","author":"Devuyst","year":"2010","journal-title":"J. Am. Soc. Nephrol"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"514","DOI":"10.4049\/jimmunol.179.1.514","article-title":"Nod1\/RICK and TLR signaling regulate chemokine and antimicrobial innate immune responses in mesothelial cells","volume":"179","author":"Park","year":"2007","journal-title":"J. Immunol"},{"key":"ref_6","first-page":"1289","article-title":"Endotoxin-induced chemokine expression in murine peritoneal mesothelial cells: The role of toll-like receptor 4","volume":"15","author":"Kato","year":"2004","journal-title":"J. Am. Soc. Nephrol"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"684","DOI":"10.1038\/ni.1606","article-title":"Sequential control of Toll-like receptor-dependent responses by IRAK1 and IRAK2","volume":"9","author":"Kawagoe","year":"2008","journal-title":"Nat. Immunol"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.immuni.2008.06.004","article-title":"When signaling pathways collide: Positive and negative regulation of Toll-like receptor signal transduction","volume":"29","year":"2008","journal-title":"Immunity"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1002\/eji.200838161","article-title":"IRAK4 in TLR\/IL-1R signaling: Possible clinical applications","volume":"38","author":"Li","year":"2008","journal-title":"Eur. J. Immunol"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"344","DOI":"10.1016\/j.cell.2008.01.020","article-title":"Shared principles in NF-\u03baB signaling","volume":"132","author":"Hayden","year":"2008","journal-title":"Cell"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1101\/gad.13.8.1015","article-title":"TRAF6 deficiency results in osteopetrosis and defective interleukin-1, CD40, and LPS signaling","volume":"13","author":"Lomaga","year":"1999","journal-title":"Genes Dev"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1155\/1998\/69628","article-title":"CD40 in clinical inflammation: From multiple sclerosis to atherosclerosis","volume":"6","author":"Laman","year":"1998","journal-title":"Dev. Immunol"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"602","DOI":"10.1159\/000098149","article-title":"Peroxisome proliferator-activated receptor-gamma is expressed by rat peritoneal mesothelial cells: Its potential role in peritoneal cavity local defense","volume":"26","author":"Zhang","year":"2006","journal-title":"Am. J. Nephrol"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1007\/s00011-009-0012-z","article-title":"Angiotensin II upregulates Toll-like receptor 4 and enhances lipopolysaccharide-induced CD40 expression in rat peritoneal mesothelial cells","volume":"58","author":"Wu","year":"2009","journal-title":"Inflamm. Res"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4645","DOI":"10.4049\/jimmunol.179.7.4645","article-title":"A novel mechanism for TNFR-associated factor 6-dependent CD40 signaling","volume":"179","author":"Rowland","year":"2007","journal-title":"J. Immunol"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1016\/j.it.2009.05.007","article-title":"The ubiquitin-editing enzyme A20 (TNFAIP3) is a central regulator of immunopathology","volume":"30","author":"Vereecke","year":"2009","journal-title":"Trends Immunol"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8217","DOI":"10.1074\/jbc.R800032200","article-title":"A20: Central gatekeeper in inflammation and immunity","volume":"284","author":"Coornaert","year":"2009","journal-title":"J. Biol. Chem"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2350","DOI":"10.1126\/science.289.5488.2350","article-title":"Failure to regulate TNF-induced NF-kappaB and cell death responses in A20-deficient mice","volume":"289","author":"Lee","year":"2000","journal-title":"Science"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1488","DOI":"10.4049\/jimmunol.0900163","article-title":"A20 attenuates allergic airway inflammation in mice","volume":"183","author":"Kang","year":"2009","journal-title":"J. Immunol"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2313","DOI":"10.1002\/art.27545","article-title":"A20 suppresses inflammatory responses and bone destruction in human fibroblast-like synoviocytes and in mice with collagen-induced arthritis","volume":"62","author":"Hah","year":"2010","journal-title":"Arthritis Rheum"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"e46515","DOI":"10.1371\/journal.pone.0046515","article-title":"A20 (TNFAIP3) alleviates CVB3-induced myocarditis via inhibiting NF-\u03baB signaling","volume":"7","author":"Gui","year":"2012","journal-title":"PLoS One"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3715","DOI":"10.1016\/j.molimm.2007.04.002","article-title":"Sensitivity of TLR4- and -7-induced NFkappa B1 p105-TPL2-ERK pathway to TNF-receptor-associated-factor-6 revealed by RNAi in mouse macrophages","volume":"44","author":"Loniewski","year":"2007","journal-title":"Mol. Immunol"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1111\/j.1365-2249.2009.04048.x","article-title":"A20 is an early responding negative regulator of Toll-like receptor 5 signalling in intestinal epithelial cells during inflammation","volume":"159","author":"Oshima","year":"2010","journal-title":"Clin. Exp. Immunol"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1084\/jem.20071108","article-title":"Homeostatic MyD88 dependent signals cause lethal inflammation in the absence of A20","volume":"205","author":"Turer","year":"2008","journal-title":"J. Exp. Med"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2093","DOI":"10.1101\/gad.1680708","article-title":"Encoding NF-kappaB temporal control in response to TNF: Distinct roles for the negative regulators IkappaBalpha and A20","volume":"22","author":"Werner","year":"2008","journal-title":"Genes Dev"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"7905","DOI":"10.1074\/jbc.M110.182873","article-title":"Endotoxin tolerance impairs IL-1 receptor-associated kinase (IRAK) 4 and TGF-beta-activated kinase 1 activation, K63-linked polyubiquitination and assembly of IRAK1, TNF receptor-associated factor 6, and IkappaB kinasegamma and increases A20 expression","volume":"286","author":"Xiong","year":"2011","journal-title":"J. Biol. Chem"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"758","DOI":"10.1038\/ncb0805-758","article-title":"Ubiquitin signalling in the NF-kappaB pathway","volume":"7","author":"Chen","year":"2005","journal-title":"Nat. Cell Biol"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1016\/j.jnutbio.2012.04.017","article-title":"TRAF6 is functional in inhibition of TLR4-mediated NF-\u03baB activation by resveratrol","volume":"24","author":"Jakus","year":"2013","journal-title":"J. Nutr. Biochem"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3242","DOI":"10.1074\/jbc.M110.150300","article-title":"Role of the A20-TRAF6 axis in lipopolysaccharide-mediated osteoclastogenesis","volume":"286","author":"Mabilleau","year":"2011","journal-title":"J. Biol. Chem"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"986","DOI":"10.4161\/auto.6.7.13288","article-title":"Traf6 and A20 differentially regulate TLR4-induced autophagy by affecting the ubiquitination of Beclin 1","volume":"6","author":"Shi","year":"2010","journal-title":"Autophagy"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1042\/BJ20071399","article-title":"Structure of the A20 OTU domain and mechanistic insights into deubiquitination","volume":"409","author":"Komander","year":"2008","journal-title":"Biochem. J"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"526","DOI":"10.1016\/j.jmb.2007.11.092","article-title":"Molecular basis for the unique deubiquitinating activity of the NF-kappaB inhibitor A20","volume":"376","author":"Lin","year":"2008","journal-title":"J. Mol. Biol"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1038\/emboj.2008.5","article-title":"Inflammatory cardiac valvulitis in TAX1BP1-deficient mice through selective NF-kappaB activation","volume":"27","author":"Iha","year":"2008","journal-title":"EMBO J"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"240","DOI":"10.1177\/089686080502500307","article-title":"CD40 ligand expression correlates with resolution of peritonitis and mononuclear cell recruitment","volume":"25","author":"Glik","year":"2005","journal-title":"Perit. Dial. Int"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/j.immuni.2010.07.017","article-title":"The ubiquitin modifying enayme A20 restricts B cell survival and prevents autoimmunity","volume":"33","author":"Tavares","year":"2010","journal-title":"Immunity"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/S0898-6568(01)00160-7","article-title":"The TNF-receptor-associated factor family: Scaffold mole-cules for cytokine receptors, kinases and theirregulators","volume":"13","author":"Wajant","year":"2001","journal-title":"Cell Signal"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1038\/nri1184","article-title":"Signalling pathways of the TNF superfamily: A double-edged sword","volume":"3","author":"Aggarwal","year":"2003","journal-title":"Nat. Rev. Immunol"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1242\/jcs.115.4.679","article-title":"All TRAFs are not created equal: Common and distinct molecular mechanisms of TRAF-mediated signal transduction","volume":"115","author":"Chung","year":"2002","journal-title":"J. Cell Sci"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1189\/jlb.72.1.19","article-title":"Mechanisms of TNF receptor-associated factor (TRAF) regulation in B lymphocytes","volume":"72","author":"Bishop","year":"2002","journal-title":"J. Leukoc. Biol"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"28745","DOI":"10.1074\/jbc.271.46.28745","article-title":"Identification of TRAF6, a novel tumor necrosis factor receptor-associated factor protein that mediates signaling from an amino-terminal domain of the CD40 cytoplasmic region","volume":"271","author":"Ishida","year":"1996","journal-title":"J. Biol. Chem"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"3114","DOI":"10.1182\/blood-2005-02-0759","article-title":"LPS induces CD40 gene expression through the activation of NF-kappaB and STAT-1alpha in macrophages and microglia","volume":"106","author":"Qin","year":"2005","journal-title":"Blood"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/15\/4\/6592\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:10:25Z","timestamp":1760217025000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/15\/4\/6592"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,4,17]]},"references-count":41,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2014,4]]}},"alternative-id":["ijms15046592"],"URL":"https:\/\/doi.org\/10.3390\/ijms15046592","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,4,17]]}}}