Journal List > J Bacteriol Virol > v.46(3) > 1034218

Ali, Manzoor, and Koh: 3-Hydroxy-4,7-megastigmadien-9-one, Isolated from Ulva pertusa Kjellman, Inhibits LPS-Induced Inflammatory Response by Down-Regulating Mitogen-Activated Protein Kinase and NF-κB Pathways


In the present study we evaluated the anti-inflammatory potential of 3-hydroxy-4,7-megastigmadien-9-one (Comp) isolated from Ulva pertusa Kjellman, in LPS-stimulated bone marrow-derived dendritic cells (BMDCs). Comp treatment exhibited strong dose dependent inhibition of IL-12 p40 and IL-6 cytokine production with IC50 values of 7.85 ± 0.32 and 7.86 ± 0.18, respectively in LPS-stimulated BMDCs. Treatment of Comp inhibited MAPKs and NF-κB pathways in LPS-stimulated BMDCs by inhibiting the phosphorylation of ERK1/2, JNK1/2, p38 and IκB. Thus, these results suggest that Comp have a significant anti-inflammatory property and affirm further studies concerning the potentials of Comp for medicinal use.


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Figure 1.
Effects of Comp on cell viability of BMDCs. BMDCs were treated with Comp (1~50 μM) for 18 h and viability was measured using MTT assay. Data are representative of three independent experiments.
Figure 2.
Inhibitory effects of Comp on pro-inflammatory cytokine production in LPS-stimulated BMDCs. BMDCs were treated with Comp at the indicated doses for 1 h before stimulation with LPS (10 ng/ml). Enzyme-linked immunosorbent assay (ELISA) was used to measure the concentrations of murine IL-12 p40 (A), IL-6 (B), and TNF-α (C) in the culture supernatants. Data are representative of three independent experiments. Comp, 3-hydroxy-4,7-megastigmadien-9-one. ∗ p < 0.05, ∗∗ p < 0.01 vs. Comp-untreated cells in the presence of LPS.
Figure 3.
Effects of Comp on the phosphorylation of MAPK and IκBα in LPS-stimulated BMDCs. (A) Cells were pre-treated with or without Comp (50 μM) for 1 h before stimulation with LPS (10 ng/ml). Total cell lysate was obtained at the indicated time intervals. Western blot analysis was performed on the cell lysate to assess phosphorylation of ERK, JNK, p38 and IκBα. β-actin was taken as the loading control. Data are representative of three independent experiments. (B) Phosphorylation of ERK, JNK, p38 and IκBα was quantified using scanning densitometry, and the band intensities were normalized by that of β-actin. Comp, 3-hydroxy-4,7-megastigmadien-9-one. ∗ p < 0.05 vs. Comp-untreated cells in the presence of LPS.
Table 1.
Anti-inflammatory effects of Comp on LPS-stimulated BMDCs
Compound IC50 values (μM)a
IL-12 p40 IL-6 TNF-α
Compa 7.85 ± 0.32 7.86 ± 0.18 > 100
SB203580b 5.00 ± 0.21 3.50 ± 0.13 7.20 ± 0.26

a IC50 values for Comp are given in column IL-6, IL-12 p40, and TNF-α. Values >100 μM are considered to be inactive.

b Positive control.

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