Nat Prod Sci.  2018 Mar;24(1):13-20. 10.20307/nps.2018.24.1.13.

Estragole Exhibits Anti-inflammatory Activity with the Regulation of NF-κB and Nrf-2 Signaling Pathways in LPS-induced RAW 264.7 cells

• Roy A
• Park HJ
• Jung HA
• Choi JS

• KMID: 2409604
• DOI: http://doi.org/10.20307/nps.2018.24.1.13

Abstract

Estragole is a naturally occurring phenylpropanoid obtained from essential oils found in a broad diversity of plants. Although the phenylpropanoids show many biological activities, clear regulation of the inflammatory signaling pathways has not yet been determined. Here, we scrutinized the anti-inflammatory effect of estragole. The anti-inflammatory effect of estragole was determined through the inhibitory mechanisms of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), nuclear factor kappa B (NF-κB), and mitogen-activated protein kinases (MAPK) pathways and the activation of nuclear factor erythroid 2-related factor 2 (Nrf-2)/heme oxygenase (HO)-1 pathways in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Estragole significantly inhibited NO production, iNOS and COX-2 expression as well as LPS-induced NF-κB and MAPK activation. Furthermore, estragole suppressed LPS-induced intracellular ROS production but up-regulated the stress response gene HO-1 via the activation of transcription factor Nrf-2. These findings demonstrate that estragole inhibits the LPS-induced expression of inflammatory mediators via the down-regulation of iNOS, COX-2, NF-κB, and MAPK pathways, as well as the up-regulation of the Nrf-2/HO-1 pathway, indicating that this phenylpropanoid has potential therapeutic and preventive applications in various inflammatory diseases.

Keyword

Essential oils; Estragole; Anti-inflammation; Nrf-2; HO-1; RAW 264.7 cells

MeSH Terms

Down-Regulation
Mitogen-Activated Protein Kinases
NF-kappa B
Nitric Oxide Synthase Type II
Oils, Volatile
Prostaglandin-Endoperoxide Synthases
RAW 264.7 Cells*
Transcription Factors
Up-Regulation
Mitogen-Activated Protein Kinases
NF-kappa B
Nitric Oxide Synthase Type II
Oils, Volatile
Prostaglandin-Endoperoxide Synthases
Transcription Factors

Figure

• 

  Fig. 1. Inhibitory effect of estragole on the production of nitric oxide (NO) in LPS-stimulated RAW264.7 cells. Cells were pretreated with different concentrations of estragole for 2 h and stimulated with LPS (1 μg/mL) for 24 h. NO production was measured by Griess reaction. Data are presented as the mean ± standard deviation of three independent experiments.###p < 0.001 indicates significant differences from the control group. ∗p <0.05, ∗∗p <0.01, and ∗∗∗p <0.001 indicate significant differences from the LPS-treated group.

• 

  Fig. 2. Inhibitory effect of estragole on the expression of iNOS and COX-2 in LPS-stimulated RAW264.7 cells. Cells were pretreated with the indicated concentration of estragole for 2 h and stimulated with LPS (1 μg/mL) for 18 h. Data was detected by Western blot analysis with the designated antibodies. β-Actin was used as an internal control. The results presented are representative of three independent experiments. #p < 0.05 indicates significant differences from the control group. ∗p < 0.05 indicates significant differences from the LPS-treated group.

• 

  Fig. 3. Inhibitory effect of estragole on the expression of NF-κB (total protein) in LPS-stimulated RAW264.7 cells. Cells were pretreated with the indicated concentrations of estragole for 2 h and stimulated with LPS (1 μg/mL) for 18 h. Data was detected by Western blot analysis with the designated antibodies. β-Actin was used as an internal control. The results presented are representative of three independent experiments. #p <0.05 indicates significant differences from the control group. ∗p < 0.05 indicates significant differences from the LPS-treated group.

• 

  Fig. 4. Inhibitory effect of estragole on the expression of MAPKs in LPS-stimulated RAW264.7 cells. Cells were pretreated with the indicated concentrations of estragole for 2 h and stimulated with LPS (1 μg/mL) for 18 h. Data was detected by Western blot analysis with the designated antibodies. β-Actin was used as an internal control. The results presented are representative of three independent experiments. #p <0.05 indicates significant differences from the control group. ∗p < 0.05 indicates significant differences from the LPS-treated group.

• 

  Fig. 5. Inhibitory effect of estragole on the production of ROS in LPS-stimulated RAW264.7 cells. Cells pretreated with different concentrations of estragole for 2 h were stimulated with LPS (1 μg/mL) for 24 h. ROS levels were measured by DCFH-DA with fluorescent analysis. Data are presented as the mean ± standard deviation of three independent experiments. ###p < 0.01 indicates significant differences from the control group. ∗p < 0.05, ∗∗p <0.01, and ∗∗∗p <0.001 indicate significant differences from the LPS-treated group.

• 

  Fig. 6. Inhibitory effect of estragole on the expression of HO-1 and Nrf-2 in LPS-stimulated RAW264.7 cells. Cells were pretreated with the indicated concentration of estragole for 2 h and stimulated with LPS (1 μg/mL) for 18 h. Data was detected by Western blot analysis with the designated antibodies. β-Actin was used as an internal control. The results presented were representative of three independent experiments. #p < 0.05 indicates significant differences from the control group. ∗p < 0.05 indicates significant differences from the LPS-treated group.

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