(E)-3-(3-methoxyphenyl)-1-(2-pyrrolyl)-2-propenone displays suppression of inflammatory responses via inhibition of Src, Syk, and NF-κB

Abstract

(E)-3-(3-methoxyphenyl)-1-(2-pyrrolyl)-2-propenone (MPP) is an aldol condensation product resulting from pyrrole-2-carbaldehyde and m- and p- substituted acetophenones. However, its biological activity has not yet been evaluated. Since it has been reported that some propenone-type compounds display anti-inflammatory activity, we investigated whether MPP could negatively modulate inflammatory responses. To do this, we employed lipopolysaccharide (LPS)-stimulated macrophage-like RAW264.7 cells and examined the inhibitory levels of nitric oxide (NO) production and transcriptional activation, as well as the target proteins involved in the inflammatory signaling cascade. Interestingly, MPP was found to reduce the production of NO in LPS-treated RAW264.7 cells, without causing cytotoxicity. Moreover, this compound suppressed the mRNA levels of inflammatory genes, such as inducible NO synthase (iNOS) and tumor necrosis factor (TNF)-α. Using luciferase reporter gene assays performed in HEK293 cells and immunoblotting analysis with nuclear protein fractions, we determined that MPP reduced the transcriptional activation of nuclear factor (NF)-κB. Furthermore, the activation of a series of upstream signals for NF-κB activation, composed of Src, Syk, Akt, and IκBα, were also blocked by this compound. It was confirmed that MPP was able to suppress autophosphorylation of overexpressed Src and Syk in HEK293 cells. Therefore, these results suggest that MPP can function as an anti-inflammatory drug with NF-κB inhibitory properties via the suppression of Src and Syk.

Src Is a Prime Target Inhibited by Celtis choseniana Methanol Extract in Its Anti‐Inflammatory Action

“…To examine the effects of Cc-ME on the activity of upstream molecules for the NF- κ B signaling pathway, we determined the phosphorylation of those upstream molecules in LPS-treated RAW264.7 cells after Cc-ME treatment using Western blot analysis and immunoprecipitation. Since I κ B α , IKK α / β , AKT, p85, and Src are known as upstream regulators of NF- κ B translocation into the nucleus, the activation of these molecules was assessed by determining their phosphorylation state using their phospho-specific antibodies as reported previously [ 40 , 41 ]. Phosphorylation of I κ B α and IKK α / β was inhibited by Cc-ME from 30 to 60 min, and phosphorylation of AKT and p85 was suppressed by Cc-ME from 5 to 60 min in the LPS-treated RAW264.7 cells (Figures 3(a) and 3(b) ).…”

Pyrrole-Derivative of Chalcone, (E)-3-Phenyl-1-(2-Pyrrolyl)-2-Propenone, Inhibits Inflammatory Responses via Inhibition of Src, Syk, and TAK1 Kinase Activities

“…Semiquantitative RT reactions were conducted as previously reported (Lee et al ., 2014b). The primers (Bioneer, Seoul, Republic of Korea) used in these experiments are the same as previously published paper (Kim et al ., 2016). …”

JS-III-49, a hydroquinone derivative, exerts anti-inflammatory activity by targeting Akt and p38

“…Therefore, intracellular signaling molecules produced during inflammatory responses are potential pharmaceutical targets to prevent and treat inflammatory diseases. Researchers have therefore actively focused on identifying and validating novel anti-inflammatory natural products such as saponins, flavonoids, and anthraquinones as well as chemical agents such as (E)-3-(3-methoxyphenyl)-1-(2-pyrrolyl)-2-propenone that specifically target intracellular signaling molecules in macrophages [ 10 11 12 13 14 15 ].…”