Inhibition of lipopolysaccharide-induced inducible nitric oxide synthase and cyclooxygenase-2 gene expression by 5-aminoimidazole-4-carboxamide riboside is independent of AMP-activated protein kinase

• Published in: Journal of cellular biochemistry 2008-02, Vol.103 (3), p.931-940
• Main Authors: Kuo, Chih-Lin, Ho, Feng-Ming, Chang, Mei Ying, Prakash, Ekambaranellore, Lin, Wan-Wan
• Format: Article
• Language: English
• Subjects: Adenosine Kinase - metabolism; AICAR; Aminoimidazole Carboxamide - analogs & derivatives; Aminoimidazole Carboxamide - metabolism; AMP-Activated Protein Kinases; AMPK; Animals; Anti-Inflammatory Agents - pharmacology; COX-2; Cyclooxygenase 2 - metabolism; Electrophoretic Mobility Shift Assay; Enzyme Activation - physiology; Gene Expression; Genes, Reporter; Inflammation - chemically induced; Inflammation - drug therapy; Inflammation - physiopathology; iNOS; Lipopolysaccharides - metabolism; macrophages; Macrophages - enzymology; Mice; Microglia - enzymology; Multienzyme Complexes - metabolism; NF-κB; Nitric Oxide Synthase Type II - antagonists & inhibitors; Nitric Oxide Synthase Type II - metabolism; Protein Binding; Protein-Serine-Threonine Kinases - metabolism; Ribonucleosides - metabolism; RNA, Messenger - metabolism; Signal Transduction; Transcription Factors - metabolism; Transcription, Genetic
• ISSN: 0730-2312; 1097-4644
• DOI: 10.1002/jcb.21466

Recent studies suggest AMP‐activated protein kinase (AMPK), an enzyme involved in energy homeostasis, might be a novel signaling pathway in regulating inflammatory response, but the precise intracellular mechanisms are not fully understood. In this study, we have demonstrated that 5‐aminoimidazole‐4‐carboxamide riboside (AICAR), an activator of AMPK, inhibited lipopolysaccharide (LPS)‐induced protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2) in macrophages and microglial cells at the gene transcription level. Data obtained from electrophoretic mobility shift assay (EMSA) and promoter activity assay have further confirmed the ability of AICAR to block LPS‐mediated NF‐κB, AP‐1, CREB, and C/EBPβ activation. However, AICAR did not affect LPS‐mediated IKK, ERK, and p38 activation. Regardless of the ability of AICAR to activate AMPK, the inhibitory effects of AICAR on iNOS and COX‐2 expression were not associated with AMPK. An adenosine kinase inhibitor 5′‐iodotubercidin, which effectively abolished AMPK activation caused by AICAR, did not reverse the anti‐inflammatory effect of AICAR. Moreover, another AMPK activator metformin was not able to mimic the effects of AICAR. Direct addition of AICAR in EMSA assay interrupted binding of NF‐κB, CREB, and C/EBPβ to specific DNA elements. Taken together, this study demonstrates that the anti‐inflammatory effects of AICAR against LPS‐induced iNOS and COX‐2 gene transcription are not associated with AMPK activation, but might be resulting from the direct interference with DNA binding to transcription factors. J. Cell. Biochem. 103: 931–940, 2008. © 2007 Wiley‐Liss, Inc.