Synergism and Subadditivity of Verbascoside-Lignans and -Iridoids Binary Mixtures Isolated from Castilleja tenuiflora Benth. on NF-κB/AP-1 Inhibition Activity
Abstract
:1. Introduction
2. Results
2.1. Isolation and Identification of Compounds
2.2. EC50 and Emax of the Isolated Compounds
2.3. Pharmacodynamic Screening Using Binary Mixtures
2.4. Pharmacodynamic Interactions
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Extraction and Isolation of Chemical Compounds from Methanolic Extracts
4.3. TLC and HPLC Analysis
4.4. UPLC Analysis
4.5. 1H- and 13C-NMR Experiments
4.6. Raw-Blue™ Cells Culture Experiments
- IA: Inhibition activity (%),
- A: Absorbance of 100% activity,
- B: Absorbance of the treatment.
4.6.1. Cell Viability on Raw-Blue ™ Cells Using the MTT Method
- C: Absorbance of the control,
- T: Treatment absorbance.
4.6.2. Inhibition of NF-κB/AP-1 by the Effective Concentration 50 (EC50) of Compounds Isolated from C. tenuiflora Using Raw-Blue™ Cells
4.6.3. Inhibition of NF-κB/AP-1 in LPS Stimulated Raw-blue ™ Cells by Binary Mixtures of Iridoid Glycosides and Lignans with Verbascoside
4.6.4. Pharmacodynamic Interaction of Binary Mixtures of Iridoids and Lignans with Verbascoside in RAW Blue™ Cells
- OIA: Observed inhibition activity (%),
- TIA: Expected inhibition activity (%).
4.7. Statistical Analyses
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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Compound | Emax (% Inhibition) | EC50 (µM) |
---|---|---|
Verbascoside | 55.55 | 10.08 |
Aucubin | 40.00 | 0.08 |
Geniposide | 57.47 | 0.01 |
Tenuifloroside | 52.91 | 0.11 |
Magnolin | 60.97 | 3.05 |
Eudesmin | 56.82 | 0.006 |
Kobusin | 45.45 | 2.37 |
Sesamin | 52.63 | 0.01 |
Mixture | Expected NF-κB/AP-1 Inhibition (%) | Observed NF-κB/AP-1 Inhibition (%) | Inhibition Ratio |
---|---|---|---|
Indomethacin | 36.31 ± 9.15 a | ||
Verbascoside-aucubin | 53.65 | 46.16 ± 3.67 a,b | 0.84 |
Verbascoside-geniposide | 48.58 | 43.04 ± 5.59 a,b | 0.89 |
Verbascoside-tenuifloroside | 60.18 | 37.99 ± 7.70 a | 0.63 |
Verbascoside-magnolin | 52.95 | 39.33 ± 7.59 a,b | 0.74 |
Verbascoside-eudesmin | 52.70 | 51.01 ± 7.99 b | 0.97 |
Verbascoside-kobusin | 55.60 | 63.06 ± 5.90 b,c | 1.13 |
Verbascoside-sesamin | 44.94 | 47.40 ± 16.28 b,c | 1.05 |
Binary Mixture | Expected NF-κB/AP-1 Inhibition (%) | Observed NF-κB/AP-1 Inhibition (%) | Inhibition Ratio |
---|---|---|---|
V-A proportion Indomethacin | 33.22 ± 6.53 a | ||
1.00/0:00 | 26.54 | 26.10 ± 4.24 a | 1.05 |
0.75/0.25 | 27.53 | 34.82 ± 8.37 a | 1.32 |
0.50/0.50 | 28.52 | 34.21 ± 4.15 a | 1.21 |
0.25/0.75 | 29.52 | 25.47 ± 10.65 a | 0.86 |
0.00/1.00 | 29.84 | 30.42 ± 4.20 a | 1.02 |
V-K proportion | |||
Indomethacin | 34.07 ± 9.13 b,c | ||
1.00/0:00 | 26.54 | 22.77 ± 5.57 a,b | 0.91 |
0.75/0.25 | 27.46 | 33.93 ± 4.07 b,c | 1.24 |
0.50/0.50 | 28.38 | 43.23 ± 5.17 c | 1.56 |
0.25/0.75 | 29.29 | 36.52 ± 4.60 b,c | 1.26 |
0.00/1.00 | 30.21 | 29.10 ± 6.63 a | 0.95 |
V-T proportion | |||
Indomethacin | 30.77 ± 4.91 b,c | ||
1.00/0:00 | 26.54 | 26.45 ± 2.47 b | 0.93 |
0.75/0.25 | 28.60 | 20.59 ± 2.70 ª | 0.72 |
0.50/0.50 | 30.66 | 14.16 ± 6.07 ª | 0.46 |
0.25/0.75 | 32.72 | 28.66 ± 2.82 b | 0.91 |
0.00/1.00 | 34.78 | 37.98 ± 5.97 c | 1.07 |
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Arango-De la Pava, L.D.; Zamilpa, A.; Trejo-Espino, J.L.; Domínguez-Mendoza, B.E.; Jiménez-Ferrer, E.; Pérez-Martínez, L.; Trejo-Tapia, G. Synergism and Subadditivity of Verbascoside-Lignans and -Iridoids Binary Mixtures Isolated from Castilleja tenuiflora Benth. on NF-κB/AP-1 Inhibition Activity. Molecules 2021, 26, 547. https://doi.org/10.3390/molecules26030547
Arango-De la Pava LD, Zamilpa A, Trejo-Espino JL, Domínguez-Mendoza BE, Jiménez-Ferrer E, Pérez-Martínez L, Trejo-Tapia G. Synergism and Subadditivity of Verbascoside-Lignans and -Iridoids Binary Mixtures Isolated from Castilleja tenuiflora Benth. on NF-κB/AP-1 Inhibition Activity. Molecules. 2021; 26(3):547. https://doi.org/10.3390/molecules26030547
Chicago/Turabian StyleArango-De la Pava, Luis David, Alejandro Zamilpa, José Luis Trejo-Espino, Blanca Eda Domínguez-Mendoza, Enrique Jiménez-Ferrer, Leonor Pérez-Martínez, and Gabriela Trejo-Tapia. 2021. "Synergism and Subadditivity of Verbascoside-Lignans and -Iridoids Binary Mixtures Isolated from Castilleja tenuiflora Benth. on NF-κB/AP-1 Inhibition Activity" Molecules 26, no. 3: 547. https://doi.org/10.3390/molecules26030547