Synthetic Fluororutaecarpine Inhibits Inflammatory Stimuli and Activates Endothelial Transient Receptor Potential Vanilloid-Type 1
"> Figure 1
<p>Effects of 10-fluoro-2-methoxyrutaecarpine (F-RUT) on nitric oxide (NO) and tumor necrosis factor (TNF)-α release by lipopolysaccharide (LPS)-treated (40 ng/mL) RAW264.7 macrophages. (<b>a</b>) NO levels were detected in culture medium using the Griess reaction; (<b>b</b>) TNF-α release in cell supernatants was detected using a mouse TNF-α Quantikine kit; (<b>c</b>) Cell viability upon F-RUT and rutaecarpine (RUT) treatment for 24 h in an MTT assay. Values are expressed as the mean ± SE. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01.</p> "> Figure 1 Cont.
<p>Effects of 10-fluoro-2-methoxyrutaecarpine (F-RUT) on nitric oxide (NO) and tumor necrosis factor (TNF)-α release by lipopolysaccharide (LPS)-treated (40 ng/mL) RAW264.7 macrophages. (<b>a</b>) NO levels were detected in culture medium using the Griess reaction; (<b>b</b>) TNF-α release in cell supernatants was detected using a mouse TNF-α Quantikine kit; (<b>c</b>) Cell viability upon F-RUT and rutaecarpine (RUT) treatment for 24 h in an MTT assay. Values are expressed as the mean ± SE. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01.</p> "> Figure 2
<p>Effect of 10-fluoro-2-methoxyrutaecarpine (F-RUT) on inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expressions by lipopolysaccharide (LPS)-treated RAW264.7 macrophages (<b>a</b>), and luciferase reporter plasmid-transfected macrophages (<b>b</b>). Cells were transfected with 2.5 μg of the pGL4.32 [luc2P/NF-κB-RE/Hygro] reporter plasmid, then treated with different concentrations of F-RUT and LPS (40 ng/mL) for 24 h. Levels of luciferase activity were determined as described in Materials and Methods. Values are expressed as the mean ± SE of triplicate tests. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01 versus LPS treatment.</p> "> Figure 3
<p>Effects of 10-fluoro-2-methoxyrutaecarpine (F-RUT) on migration and invasion. Cell migration (<b>a</b>) and cell invasion (<b>b</b>) were detected following F-RUT treatment for 0–24 h, and photographed with a microscope (upper panel). The statistical analysis is shown in the lower panel. (* <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01).</p> "> Figure 4
<p>Effects of 10-fluoro-2-methoxyrutaecarpine (F-RUT) on transient receptor potential vanilloid-type 1 (TRPV-1) expression and endothelial nitric oxide synthase (eNOS) phosphorylation in human aortic endothelial cells (HAECs). The densitometric ratio is indicated.</p> "> Figure 5
<p>Effects of 10-fluoro-2-methoxyrutaecarpine (F-RUT) on ovalbumin (OVA)-challenged mice. Data are representative of three to five mice per group. Scale bar is 100 µm.</p> "> Scheme 1
<p>Synthesis of 10-fluoro-2-methoxyrutaecarpine (F-RUT) and 10-fluoro-2,3-dimethoxyrutaecarpine.</p> ">
Abstract
:1. Introduction
2. Results
2.1. Design and Synthesis of F-RUT
2.2. NO and TNF-Alpha Releases by LPS-Treated Macrophages Were Suppressed by F-RUT
2.3. F-RUT Suppresses Inducible (i)NOS and COX-2 Expressions which Was Correlated with Inhibition of Nuclear Factor (NF)-κB Activity in LPS-Activated Macrophages
2.4. F-RUT Inhibited Cell Migration/Invasion
2.5. F-RUT Activated TRPV1 and eNOS
2.6. F-RUT Ameliorates Inflammation in OVA/Alum-Challenged Mice
3. Discussion
4. Materials and Methods
4.1. Chemicals and General Methods
4.2. Synthesis of 10-Fluoro-2-methoxyrutaecarpine (FMO-RUT)
4.3. Cell Culture
4.4. Nitrite Assay
4.5. TNF-α Assay
4.6. Cell Viability Assay
4.7. Western Blot Analysis
4.8. Transient Transfection and Luciferase Assay
4.9. Wound-Healing Migration Assay
4.10. In Vitro Invasion Assays
4.11. Animal Experiment
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample Availability: Sample of the F-RUT is available from the authors. |
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Lee, C.-M.; Gu, J.-A.; Rau, T.-G.; Wang, C.; Yen, C.-H.; Huang, S.-H.; Lin, F.-Y.; Lin, C.-M.; Huang, S.-T. Synthetic Fluororutaecarpine Inhibits Inflammatory Stimuli and Activates Endothelial Transient Receptor Potential Vanilloid-Type 1. Molecules 2017, 22, 656. https://doi.org/10.3390/molecules22040656
Lee C-M, Gu J-A, Rau T-G, Wang C, Yen C-H, Huang S-H, Lin F-Y, Lin C-M, Huang S-T. Synthetic Fluororutaecarpine Inhibits Inflammatory Stimuli and Activates Endothelial Transient Receptor Potential Vanilloid-Type 1. Molecules. 2017; 22(4):656. https://doi.org/10.3390/molecules22040656
Chicago/Turabian StyleLee, Chi-Ming, Jiun-An Gu, Tin-Gan Rau, Chi Wang, Chiao-Han Yen, Shih-Hao Huang, Feng-Yen Lin, Chun-Mao Lin, and Sheng-Tung Huang. 2017. "Synthetic Fluororutaecarpine Inhibits Inflammatory Stimuli and Activates Endothelial Transient Receptor Potential Vanilloid-Type 1" Molecules 22, no. 4: 656. https://doi.org/10.3390/molecules22040656
APA StyleLee, C. -M., Gu, J. -A., Rau, T. -G., Wang, C., Yen, C. -H., Huang, S. -H., Lin, F. -Y., Lin, C. -M., & Huang, S. -T. (2017). Synthetic Fluororutaecarpine Inhibits Inflammatory Stimuli and Activates Endothelial Transient Receptor Potential Vanilloid-Type 1. Molecules, 22(4), 656. https://doi.org/10.3390/molecules22040656