title: G2013 modulates TLR4 signaling pathway in IRAK-1 and TARF-6 dependent and miR-146a independent manner.
abstract: Inflammation is inseparable part of different diseases especially cancer and autoimmunity. During inflammation process toll like receptor 4(TLR4) responds to lipopolysaccharide (LPS), one of the bacterial components, and TLR4 signaling leads to interleukine-1 receptor associated kinase-1 (IRAK1) and tumor necrosis factor (TNF) receptor associated factor6 (TRAF6) activation which ultimately results in nuclear factor- ĸB (NF-ĸB) activation as the main transcription factor of inflammatory cytokines. Conversely, NF-ĸB over activation induces miR-146a in innate immune cells which can consequently reduce TRAF6, IRAK1, and NF-ĸB activation in a negative feedback. G2013 is a novel designed non-steroidal anti-inflammatory drug (NSAID) which was recently shown to be effective in experimental autoimmune encephalomyelitis (EAE) mouse model. The aim of this study was to evaluate G2013 effects on inflammatory (IRAK1 and TRAF6) and anti-inflammatory (miR-146a) factors of TLR4 signaling pathway. For this purpose, cytotoxicity of G2013 has been evaluated by MTT assay. Expression level of miR-146a in PBMCs and IRAK1 along with TRAF6 in HEK-293 TLR4 cells have been determined using real time PCR. Our results showed that IC50 of G2013 was 25μg/ml, thus 5 and 25 μg/ml concentrations used for further treatments as low dose and high dose concentrations. Our results showed that IRAK1 expression reduced between 5 to 8 fold after treatment by G2013 in a dose dependent manner (p<0.001). In parallel TRAF6 expression declined between 3 to 10 fold dose dependently (p<0.05). However, miR-146a expression was not affected after treatment with low dose and high dose of G2013. In conclusion our data showed that G2013 can regulate TLR4 signaling pathway during inflammation by reducing downstream signaling molecules, IRAK1 and TRAF6 without altering miR-146a expression.
authors.affiliation: Tabriz University of Medical Sciences Hematology and Oncology Research Center Tabriz Iran.
authors.forename: M
authors.initials: M
authors.lastname: Hajivalili
authors.affiliation: Tabriz University of Medical Sciences Hematology and Oncology Research Center Tabriz Iran.
authors.forename: F
authors.initials: F
authors.lastname: Pourgholi
authors.affiliation: Tabriz University of Medical Sciences Immunology Research Center Tabriz Iran.
authors.forename: J
authors.initials: J
authors.lastname: Majidi
authors.affiliation: Tabriz University of Medical Sciences Immunology Research Center Tabriz Iran.
authors.forename: L
authors.initials: L
authors.lastname: Aghebati-Maleki
authors.affiliation: Tabriz University of Medical Sciences Hematology and Oncology Research Center Tabriz Iran.
authors.forename: A A
authors.initials: AA
authors.lastname: Movassaghpour
authors.affiliation: Tabriz University of Medical Sciences Department of Immunology, Faculty of Medicine Tabriz Iran.
authors.forename: H
authors.initials: H
authors.lastname: Samadi Kafil
authors.affiliation: Tehran University of Medical Sciences Department of Immunology, School of Public Health Tehran Iran.
authors.forename: A
authors.initials: A
authors.lastname: Mirshafiey
authors.affiliation: Tabriz University of Medical Sciences Immunology Research Center Tabriz Iran Yousefime@tbzmed.ac.ir.
authors.forename: M
authors.initials: M
authors.lastname: Yousefi
mesh_terms: D049109:Cell Proliferation / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D057809:HEK293 Cells; D006603:Hexuronic Acids / Q000494:pharmacology*; D006801:Humans; D053592:Interleukin-1 Receptor-Associated Kinases / Q000235:genetics / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D048029:TNF Receptor-Associated Factor 6 / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism*; D014574:Uronic Acids / Q000494:pharmacology*
chemical_list: D006603:Hexuronic Acids; C542542:MIRN146 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; C495345:TLR4 protein, human; D048029:TNF Receptor-Associated Factor 6; D051197:Toll-Like Receptor 4; D014574:Uronic Acids; C007896:guluronic acid; C008324:mannuronic acid; C507768:IRAK1 protein, human; D053592:Interleukin-1 Receptor-Associated Kinases