Pharmacological mechanism underlying anti-inflammatory properties of two structurally divergent coumarins through the inhibition of pro-inflammatory enzymes and cytokines

Background: the aim of the present study is to investigate the effects of two structurally divergent coumarins,calipteryxin (1) and (3's,4's)-3',4'-disenecioyloxy-3',4'-dihydroseselin (2) from seseli recinosum,in lipopolysaccharide (lps)-stimulated murine macrophages. methods: the nitrite production was evaluated using griess reagent. the protein and mrna expression levels were investigated through western blot and quantitative real time-pcr analyses. the nf-κb and ap-1 dna-binding activities were assessed using an electrophoretic mobility shift assay. the docking studies were performed with glide xp in schrödinger suite (version 2013). results: the results of the present study revealed that calipteryxin (1) and (3's,4's)-3',4'-disenecioyloxy-3',4'-dihydroseselin (2) treatment showed potent inhibitory effects on pro-inflammatory enzymes and cytokines associated with molecular signaling pathways. treatment with calipteryxin and (3's,4's)-3',4'-disenecioyloxy-3',4'-dihydroseselin also decreased the production of nitric oxide (no),tumor necrosis factor alpha (tnf-α) and interleukin-1 beta (il-1β) in a dose-dependent manner. additionally,both coumarins inhibited the lps-induced protein and mrna expression levels of nitric oxide synthase (inos) and cyclooxygenase-2 (cox-2) in raw264.7 cells. to explore the potential mechanisms underlying the inhibitory activity of coumarin derivatives,the protein signaling pathways for nf-κb,mitogen-activated protein kinase (mapk) and akt were examined. calipteryxin and (3's,4's)-3',4'-disenecioyloxy-3',4'-dihydroseselin markedly reduced the lps-stimulated phosphorylation of ikkα/β,p-iκbα and iκbα degradation as well as the nuclear translocation of the p65 subunit of pro-inflammatory transcription factor nf-κb. in addition,calipteryxin and (3's,4's)-3',4'-disenecioyloxy-3',4'-dihydroseselin) considerably inhibited the lps-induced expression of erk,c-jun n-terminal kinase (jnk),p38 and akt proteins. furthermore,both coumarins significantly inhibited c-jun expression in the nucleus. conclusions: taken together,these results support the therapeutic potential and molecular mechanism of calipteryxin and (3's,4's)-3',4'-disenecioyloxy-3',4'-dihydroseselin associated with inflammatory diseases. © 2015 khan et al.