Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Tetra-O-methyl nordihydroguaiaretic acid (Terameprocol) inhibits the NF- B-dependent transcription of TNF-a and MCP-1/CCL2 genes by preventing RelA from binding its cognate sites on DNA. | Oyegunwa et al. Journal of Inflammation 2010 7 59 http content 7 1 59 JOURNAL OF INFLAMMATION RESEARCH Open Access Tetra-O-methyl nordihydroguaiaretic acid Terameprocol inhibits the NF- B-dependent transcription of TNF-a and MCP-1 CCL2 genes by preventing RelA from binding its cognate sites on DNA Akinbolade O Oyegunwa Michael L Sikes Jason R Wilson Frank Scholle Scott M Laster Abstract Background Tetra-O-methyl nordihydroguaiaretic acid also known as terameprocol TMP is a naturally occurring phenolic compound found in the resin of the creosote bush. We have shown previously that TMP will suppress production of certain inflammatory cytokines chemokines and lipids from macrophages following stimulation with LPS or infection with H1N1 influenza virus. In this study our goal was to elucidate the mechanism underlying TMP-mediated suppression of cytokine and chemokine production. We focused our investigations on the response to LPS and the NF-kB protein RelA a transcription factor whose activity is critical to LPS-responsiveness. Methods Reporter assays were performed with HEK293 cells overexpressing either TLR-3 -4 or -8 and a plasmid containing the luciferase gene under control of an NF-kB response element. Cells were then treated with LPS poly I C or resiquimod and or TMP and lysates measured for luciferase activity. RAW cells treated with LPS and or TMP were used in ChIP and EMSA assays. For ChIP assays chromatin was prepared and complexes precipitated with anti-NF-KB RelA Ab. Cross-links were reversed DNA purified and sequence abundance determined by Q-PCR. For EMSA assays nuclear extracts were incubated with radiolabeled probes analyzed by non-denaturing PAGE and visualized by autoradiography. RAW cells treated with LPS and or TMP were also used in fluorescence microscopy and western blot experiments. Translocation experiments were performed using a primary Ab to NF-kB RelA and a fluorescein-conjugated secondary Ab. Western
