3β-Angeloyloxy-8β,10β-dihydroxyeremophila-7(11)-en-12,8α-lactone Inhibits Lipopolysaccharide-Induced Nitric Oxide Production in RAW264.7 Cells

Abstract

Farfugium japonicum (L.) KITAM, named "Lian-Peng-Cao" in China, has been traditionally used in Chinese folk medicine to treat sore throat, cold and cough due to its anti-inflammatory properties. In this study, the anti-inflammatory action of 3β-angeloyloxy-8β,10β-dihydroxyeremophila-7(11)-en-12,8α-lactone (FJ1) isolated from Farfugium japonicum and its molecular mechanism in RAW264.7 cells were investigated. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that FJ1 with or without 3 µg/mL lipopolysaccharide (LPS) had no significant cytotoxicity in RAW264.7 cells. The production of nitric oxide (NO) was identified with a Griess reagent kit. The mRNA expression of inducible nitric oxide synthase (iNOS) and cytokines, including tumor necrosis factor α (TNF-α) and cyclooxygenase-2 (COX-2), was measured by real-time polymerase chain reaction (PCR). Reactive oxygen species (ROS) production was detected by flow cytometry analysis. Western blot was used to examine the protein expression of nuclear factor-kappa B (NF-κB)/p65, inhibitor of kappa B (IκB)-α, phosphorylated IκB-α (p-IκB-α), mitogen-activated protein kinase (MAPK) molecules, iNOS, and TNF-α. We discovered that FJ1 possesses anti-inflammatory effects that inhibit the release of LPS-stimulated pro-inflammatory cytokines, including NO and ROS. The molecular mechanism of FJ1-mediated anti-inflammation is associated with decreasing phosphorylation of MAPK molecules, including extracellular signal-related kinase 1/2 (ERK1/2), p38 MAPK, and c-Jun NH2-terminal kinase (JNK), FJ1 also reverses IκB degradation and attenuates the mRNA and protein expression of NF-κB-related downstream inducible enzymes and cytokines, such as iNOS, TNF-α in RAW264.7 cells. The results suggest that FJ1 has anti-inflammatory properties, which indicates that F. japonicum can be utilized to treat inflammatory diseases. The potential mechanism is associated with the NF-κB and MAPK activation pathways in LPS-stimulated macrophages.

<i>O</i>-Methylbulbocapnine and Dicentrine Suppress LPS-Induced Inflammatory Response by Blocking NF-κB and AP-1 Activation through Inhibiting MAPKs and Akt Signaling in RAW264.7 Macrophages

“…After 4 h of treatment, the cells were incubated with or without LPS (1 µg/mL) for 24 h. The culture mediums were collected and analyzed for IL-6, TNF-α, and PGE 2 production using ELISA kits according to the manufacturer's protocol. 21,22) To determine the total concentration of NO in the culture media, 150 µL of Griess reagent (1% sulfanilamide, 0.1% N-1-naphthylenediamine dihydrochloride and 2.5% phosphoric acid) was mixed with 150 µL of the supernatant and the absorbance at 540 nm was evaluated using a microplate reader. 23) Western Blot Analysis Preparations of whole cell lysates, cytoplasmic and nuclear fractions were carried out according to the previously published procedures.…”

Development and application of an efficient recombineering system for Burkholderia glumae and Burkholderia plantarii

“…Compounds 1 – 7 were evaluated for their ability to inhibit NO production in LPS‐stimulated RAW264.7 cells using the Griess assay (Sun et al ., 2015). Notably, none of the compounds displayed cytotoxicity with murine macrophage RAW264.7 cells using the MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide, Sigma] assay (IC 50 > 48 μM; Table S14; Alley et al ., 1988), which suggested that any inhibition of NO production would not be due to cytotoxicity.…”

Obtención y actividad antiinflamatoria de flavonoides y terpenoides aislados de Cayaponia tayuya e Isodon xerophilus

“…En la búsqueda de nuevos antiinflamatorios, muchos compuestos derivados de plantas han sido investigados en su actividad antiinflamatoria in vivo y en los mecanismos de acción involucrados, centrando el foco en la inhibición de la producción de mediadores proinflamatorios y en la inhibición de la expresión de enzimas y de la activación de factores de trascripción implicados en las respuestas inflamatorias (Risitano et al, 2014;Chen et al, 2015;Choi et al, 2015;Fang et al, 2015;Hong et al, 2015;Limtrakul et al, 2015;Sun et al, 2015). (Ríos et al, 2009).…”