Suppressive effect of novel aromatic diamine compound on nuclear factor-kappaB-dependent expression of inducible nitric oxide synthase in macrophages.
N1-benzyl-4-methylbenzene-1,2-diamine (BMD) is a novel synthetic compound. In the present study, BMD compound was discovered to inhibit nitric oxide (NO) production in macrophages RAW 264.7. BMD compound attenuated lipopolysaccharide (LPS)-induced synthesis of both mRNA and protein of inducible nitric oxide synthase (iNOS), and inhibited LPS-induced iNOS promoter activity, indicating that the aromatic diamine compound could down-regulate iNOS expression at the transcription level. As a mechanism of the anti-inflammatory action, suppression of BMD compound on nuclear factor (NF)-kappaB activation has been documented. BMD compound exhibited dose-dependent inhibitory effect on LPS-mediated NF-kappaB transcriptional activity in the macrophages. Further, the compound inhibited LPS-mediated nuclear translocation of NF-kappaB p65 and DNA binding activity of NF-kappaB complex, in parallel, but did not affect LPS-mediated degradation of inhibitory kappaBalpha protein (IkappaBalpha). These results indicate that BMD compound could inhibit nuclear localization step of NF-kappaB p65 without affecting IkappaBalpha degradation. Finally, BMD compound could provide an invaluable tool to investigate NF-kappaB-dependent iNOS expression, in addition to its therapeutic potential in NO-associated inflammatory diseases.
Shin
Hyun-Mo
HM
College of Pharmacy and Research Center for Bioresource and Health, Chungbuk National University, Cheongju 361-763, Korea.
Byung Hak Kim
Eun Yong Chung
Jung
Sang-Hun
SH
Yeong Shik Kim
Kyung Rak Min
Kim
Youngsoo
Y
D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000588:Amines / Q000494:pharmacology*; D000818:Animals; D001593:Benzyl Compounds / Q000494:pharmacology*; D015153:Blotting, Western; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D004789:Enzyme Activation / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D020672:I-kappa B Proteins / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics* / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051996:Transcription Factor RelA / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D000588:Amines; D001593:Benzyl Compounds; D020672:I-kappa B Proteins; D008070:Lipopolysaccharides; C497734:N1-benzyl-4-methylbenzene-1,2-diamine; D016328:NF-kappa B; D009573:Nitrites; D009841:Oligonucleotides; D051996:Transcription Factor RelA; D009569:Nitric Oxide; D007371:Interferon-gamma; D052247:Nitric Oxide Synthase Type II; D020928:Mitogen-Activated Protein Kinases