Abstract
Background: Since the anti-inflammatory activity of arachidonic acid derivatives was previously reported, we synthesized three new amide derivatives of arachidonic acid (AA-Ds) and tested their anti-inflammatory effects on an in vitro skin inflammation model. Aim of our study was to find derivatives of natural compounds able to down regulate inflammatory signal transduction pathway.
Methods: Human keratinocytes cell line (HaCaT) was cultured and induced by cytokines in the presence of AA-Ds. Cytokines administration elicited an inflammatory response mediated by NF-κB and STAT-1 activation that induced proinflammatory genes expression.
Results: By real time PCR we found that 24 hours after induction all AA-Ds significantly inhibit inducible Nitric Oxide Synthase (iNOS), TNFα, Inhibitor α of NF-κB, chemokine (C-X-C motif) ligand 9 and 10 genes expression. We analyzed their molecular effects in particular on the iNOS gene expression. Since iNOS transcript half-life did not change with AA-Ds treatment, we excluded a prominent role of post-transcriptional regulation for this gene and focused our attention on its transcriptional regulation. Starting three-five hours after cytokines induction, HaCaT cells, pretreated with each compound, showed inhibition of both NF-κB DNA-binding and NF-κB p65-Ser536 phosphorylation. STAT1 activation was inhibited only by AA-D4 derivative. To explain why the inhibition of iNOS expression began late after induction we analyzed activities of others key transcription factors. AA-Ds treatment elicited early increases of AP1 DNA binding as well as c-Jun, c-Fos and Fra-1 mRNA levels. Our data agree with the repressing effects of AP1 on human iNOS promoter previously described in others cell systems (Kleinert et al.).
Conclusion: AA-Ds shown to be good candidates as inhibitors of several pro-inflammatory genes induction and our study provides indications for their possible use as new antiinflammatory drugs.
Keywords: Arachidonoylamide derivatives, NF-B, AP-1, STAT1, iNOS, inflammatory cytokines, Inflammation.
Graphical Abstract
Medicinal Chemistry
Title:Three Arachidonoylamide Derivatives Inhibit Pro-Inflammatory Genes Expression by Modulating NF-κB and AP1 Activities
Volume: 12 Issue: 7
Author(s): Alex Gregorelli, Anna Sgarbossa, Shahbaz Khan, Annunziata Soriente, Margherita De Rosa, Carmela Saturnino and Marta Menegazzi
Affiliation:
Keywords: Arachidonoylamide derivatives, NF-B, AP-1, STAT1, iNOS, inflammatory cytokines, Inflammation.
Abstract: Background: Since the anti-inflammatory activity of arachidonic acid derivatives was previously reported, we synthesized three new amide derivatives of arachidonic acid (AA-Ds) and tested their anti-inflammatory effects on an in vitro skin inflammation model. Aim of our study was to find derivatives of natural compounds able to down regulate inflammatory signal transduction pathway.
Methods: Human keratinocytes cell line (HaCaT) was cultured and induced by cytokines in the presence of AA-Ds. Cytokines administration elicited an inflammatory response mediated by NF-κB and STAT-1 activation that induced proinflammatory genes expression.
Results: By real time PCR we found that 24 hours after induction all AA-Ds significantly inhibit inducible Nitric Oxide Synthase (iNOS), TNFα, Inhibitor α of NF-κB, chemokine (C-X-C motif) ligand 9 and 10 genes expression. We analyzed their molecular effects in particular on the iNOS gene expression. Since iNOS transcript half-life did not change with AA-Ds treatment, we excluded a prominent role of post-transcriptional regulation for this gene and focused our attention on its transcriptional regulation. Starting three-five hours after cytokines induction, HaCaT cells, pretreated with each compound, showed inhibition of both NF-κB DNA-binding and NF-κB p65-Ser536 phosphorylation. STAT1 activation was inhibited only by AA-D4 derivative. To explain why the inhibition of iNOS expression began late after induction we analyzed activities of others key transcription factors. AA-Ds treatment elicited early increases of AP1 DNA binding as well as c-Jun, c-Fos and Fra-1 mRNA levels. Our data agree with the repressing effects of AP1 on human iNOS promoter previously described in others cell systems (Kleinert et al.).
Conclusion: AA-Ds shown to be good candidates as inhibitors of several pro-inflammatory genes induction and our study provides indications for their possible use as new antiinflammatory drugs.
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Cite this article as:
Gregorelli Alex, Sgarbossa Anna, Khan Shahbaz, Soriente Annunziata, Rosa De Margherita, Saturnino Carmela and Menegazzi Marta, Three Arachidonoylamide Derivatives Inhibit Pro-Inflammatory Genes Expression by Modulating NF-κB and AP1 Activities, Medicinal Chemistry 2016; 12 (7) . https://dx.doi.org/10.2174/1573406412666160502154936
DOI https://dx.doi.org/10.2174/1573406412666160502154936 |
Print ISSN 1573-4064 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-6638 |
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