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

Abstract

BackgroundThe 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.MethodsThe 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).ResultsThe 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.ConclusionsTaken 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12950-015-0087-y) contains supplementary material, which is available to authorized users.

ECN from <em>Tussilago Farfara</em> Attenuates Neuropathic Pain by Suppressing Oxidative Stress, Inflammatory and Pro-Apoptotic Proteins Expression in Surgical Model

“…The quantitative RT-PCR was used to determine the effect of ECN (10 mg/kg) on the mRNA expression levels of inflammatory mediators (COX-2, iNOS, TNFα, IL-1β and IL-6) and anti-oxidant proteins (Nrf2, HO-1 and NQO1) following 7 and 14 days post PSNL surgery. Trizol Reagent was used to isolate the total RNA from the sciatic nerve and lumbar dorsal spinal cord (L4-L6) tissue of mice as described previously [46,59,60]. Briefly, GenDEPOT 0.2mL 8-strip tubes were used for quantitative PCR.…”

Natural and Synthetic Coumarins with Effects on Inflammation

“…The first compound of this class extracted was seselin 27 from Sigmathantus trifoliatus able to inhibit inflammatory hyperalgesia and having also antinociceptive effect [39]. Obtained from Seseli resinosum , calipteryxin 28 and a dihydroseselin derivative were associated in the inhibition of inflammatory enzymes (iNOS and COX-2) and cytokines (TNF-α and IL-6) via NF-κB, MAPK and Akt pathways [40]. (+)-Praeruptorin 29 from the roots of Peucedanum praerupotorum reduced NO production and release of IL-1β, IL-6 and TNF-α induced by LPS-stimulated macrophage [41,42].…”

Transmission pathways and mediators as the basis for clinical pharmacology of pain

“…After establishing increases in NO in response to incision, there was decreased post-incision nociceptive behavior in rats that were treated with nitric oxide synthase (NOS) inhibitors. In a study with a similar emphasis on the importance of inflammation in the pain response, Khan et al [148] studied the effect of two distinct coumarin drugs on mediators of inflammation. Significantly, these drugs decreased the expression of NOS mRNA as well as other mediators of inflammation, including COX enzymes [148].…”

“…In a study with a similar emphasis on the importance of inflammation in the pain response, Khan et al [148] studied the effect of two distinct coumarin drugs on mediators of inflammation. Significantly, these drugs decreased the expression of NOS mRNA as well as other mediators of inflammation, including COX enzymes [148]. The findings from these studies strongly support the role of NOS as a player in the process of inflammatory pain.…”