Influence of silencing TRAF6 with shRNA on LPS/TLR4 signaling in vitro
作者： Feng Chen , Shengsong He , Rongyuan Qiu , Ran Pang , Juanjuan Xu
DOI: 10.1007/S11596-010-0343-6
关键词:
摘要: This study investigated the influence of silencing TRAF6 with shRNA on lipopolysaccharide (LPS)/toll-like receptor (TLR)-4 signaling pathway in vitro. Four plasmids (pGCsi-TRAF6-shRNA1, 2, 3, 4) containing different sequences were designed and synthesized. The proliferation RAW264.7 cells after transfected these was measured by MTT assay. Inflammatory cellular models established LPS stimulation. Levels TNF-α, IL-1β TGF-β1 supernatants, mRNA expressions TRAF6, IL-6 COX-2, protein expression translocation NF-κB assayed ELISA, real-time quantitative PCR Western blotting, respectively. results showed that gene knockdown RNAi hardly inhibited within 72 h. lower TRAF6-shRNA1, 2 groups than TRAF6-shRNA3, 4 groups. Therefore, pGCsi-TRAF6-shRNA1, selected for subsequent experiments. Our still could significantly reduce production pro-inflammatory cytokines mediators including IL-1β, inhibit nuclear translocation. Moreover, suppress release at level. It concluded recombinant plasmid pTRAF6-shRNA can, to some extent, inflammatory response stimulated initial phase. may become potential therapeutic target many inflammation-related diseases.
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