Naturally occurring flavonoids are known to modulate various inflammatory and immune processes. Based on structural property, in this study, molecular mechanism of flavonoids in modulating nitric oxide (NO) production and its signaling pathway were investigated using lipopolysaccharide (LPS)-activat...
Naturally occurring flavonoids are known to modulate various inflammatory and immune processes. Based on structural property, in this study, molecular mechanism of flavonoids in modulating nitric oxide (NO) production and its signaling pathway were investigated using lipopolysaccharide (LPS)-activated RAW264.7 cells. Although flavonol-typed flavonoids (kaempferol and quercetin) more potently scavenged reactivity of nitric oxide ($\cdot$NO) as well as peroxynitrite (ONOO$\kappa$) than isoflavones (genistein and genistin), kaempferol, quercetin and genistein showed a little difference in inhibition of both inducible NO synthase expression and NO production, with IC$_{50}$ values of 13.9, 20.1 and 26.8 $\mu$M. However, there was a striking pattern related to structural feature in modulation of LPS-mediated signaling pathways. Thus, flavonols only inhibited transcription factor AP-1 activation, whereas isoflavones suppressed the DNA binding activation of NF-$\kappa$B and C/EBP$\beta$. Therefore, these data suggest that structural feature may be linked to decide drugs target molecule in LPS-mediated signaling pathways, rather than its potency.
Naturally occurring flavonoids are known to modulate various inflammatory and immune processes. Based on structural property, in this study, molecular mechanism of flavonoids in modulating nitric oxide (NO) production and its signaling pathway were investigated using lipopolysaccharide (LPS)-activated RAW264.7 cells. Although flavonol-typed flavonoids (kaempferol and quercetin) more potently scavenged reactivity of nitric oxide ($\cdot$NO) as well as peroxynitrite (ONOO$\kappa$) than isoflavones (genistein and genistin), kaempferol, quercetin and genistein showed a little difference in inhibition of both inducible NO synthase expression and NO production, with IC$_{50}$ values of 13.9, 20.1 and 26.8 $\mu$M. However, there was a striking pattern related to structural feature in modulation of LPS-mediated signaling pathways. Thus, flavonols only inhibited transcription factor AP-1 activation, whereas isoflavones suppressed the DNA binding activation of NF-$\kappa$B and C/EBP$\beta$. Therefore, these data suggest that structural feature may be linked to decide drugs target molecule in LPS-mediated signaling pathways, rather than its potency.
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제안 방법
NOx concentration was determined by analyzing nitrite (N02) and nitrate (N03) levels using Griess assay as described in M간erial and Methods. Each experiment was performed by triplicate observations. Data are mean ± S.
데이터처리
The Student's t-test and oneway ANOVA were used to determine the statistical significance of differences between values for vari이js experimental and control groups. Data are expressed as means ± standard errors (SEM) of at least 3 independent experiments performed with triplicates.
이론/모형
Enzymatic activity of GSH red니ctase was determined by the NADPH reduction method (Mavis and St이Iwagen, 1968). The samples with 0.
0N00' scavenging activity was measured by monitoring the oxidation of DHR 123. NOx concentration was determined by analyzing nitrite (N02) and nitrate (N03) levels using Griess assay as described in M간erial and Methods. Each experiment was performed by triplicate observations.
0), with 50 mM KCI, 100 mM NaCl, 1 mM DTT, 5 mg/mL each of pepstatin and aprotinin] and incubated on ice for 30 min to obtain nuclear extracts. Protein concentrations were determined by bicinchoninic acid method (Smith et al., 1985).
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