Modulation of Oxidative Stress and Hemostasis by Flavonoids from Lentil Aerial Parts
Abstract
:1. Introduction
2. Material and Methods
2.1. Chemicals
2.2. Plant Material
2.3. Preparation of Extract and Phenolic Fraction from Lentil Aerial Parts
2.4. Isolation of Flavonoids from Lentil Aerial Parts
2.5. Stock Solutions
2.6. Quantification of Flavonoids in the Tested Crude Extract and Phenolic Fraction of the Lentil Aerial Parts
2.7. Human Plasma Isolation
- The extract from the lentil aerial parts at the final concentrations of 1–50 μg/mL;
- The phenolic fraction from lentil aerial parts at the final concentrations of 1–50 μg/mL;
- Flavonoids (compounds 1–7) from the lentil aerial parts at the final concentrations of 1–50 μg/mL;
- Quercetin or kaempferol at the final concentrations of 1–50 μg/mL;
- Human plasma was also pre-incubated (5 min, at 37 °C; for parameters of oxidative stress) with:
- The extract from the lentil aerial parts at the final concentrations of 1–50 μg/mL;
- The phenolic fraction from lentil aerial parts at the final concentrations of 1–50 μg/mL
- Flavonoids (compounds 1–7) from the lentil aerial parts at the final concentrations of 1–50 μg/mL;
- Quercetin or kaempferol at the final concentrations of 1–50 μg/mL;
- And then treated with 4.7 mM H2O2/3.8 mM Fe2SO4/2.5 mM EDTA (25 min, at 37 °C).
2.8. Markers of Oxidative Stress
2.8.1. Lipid Peroxidation Measurement
2.8.2. Carbonyl Group Measurement
2.8.3. Thiol Group Determination
2.9. Parameters of Hemostasis
2.10. Data Analysis
3. Results
3.1. Quantitative Analysis of the Tested Extract and Phenolic Fraction of the Lentil Aerial Parts
3.2. Effects on Oxidative Stress Biomarkers in Human Plasma In Vitro
3.3. Effects on Hemostatic Parameters of Plasma
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
Abbreviations
TBARS | thiobarbituric acid reactive substances |
APTT | the activated partial thromboplastin time |
PT | the prothrombin time |
TT | the thrombin time |
n.s. | statistically irrelevant |
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Compound | Preparations | |
---|---|---|
CE | PF | |
1 | 3.4 ± 0.2 | 16.5 ± 0.1 |
2 | 20.5 ± 1.3 | 103.4 ± 1.7 |
3 | 10.1 ± 0.8 | 50.7 ± 0.8 |
4 | 6.9 ± 0.5 | 35.2 ± 1.2 |
5 | 4.2 ± 0.2 | 22.0 ± 0.4 |
6 | 1.0 ± 0.1 a | 5.6 ± 0.1 a |
7 | 1.1 ± 0.1 a | 5.5 ± 0.3 a |
8 | 9.6 ± 0.6 a | 48.8 ± 0.9 a |
9 | 5.5 ± 0.4 a | 28.2 ± 0.5 a |
10 | 6.4 ± 0.4 b | 33.3 ± 1.0 b |
11 | 12.2 ± 0.8 b | 63.2 ± 1.0 b |
Tested Extract/Fraction/Phenolic Compound | Parameters of Oxidative Stress | Parameters of Hemostasis | ||
---|---|---|---|---|
Inhibition of Lipid Peroxidation Induced by H2O2/Fe (%) | Inhibition of Protein Carbonylation Induced by H2O2/Fe (%) | The Level of Protein Thiol Groups (nmol/mL of Plasma) in Plasma Treated with H2O2/Fe | Prolongation of TT (the Thrombin Time) (%) | |
Extract (a) | 19.4 ± 4.8 Antioxidant action vs. control (plasma treated with H2O2/Fe) | 6.2 ± 1.9 (p > 0.05) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 355.2 ± 13.7 Antioxidant action vs. control (plasma treated with H2O2/Fe) | No effect vs. control |
Phenolic fraction (b) | 18.2 ± 3.8 (p > 0.05; b vs. a) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 47.3 ± 7.9 (p < 0.05; b vs. a) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 263.8 ± 15.2 (p < 0.05; b vs. a) No effect vs. control (plasma treated with H2O2/Fe) | 10.8 ± 2.4 Anticoagulant action vs. control |
Quercetin (c) | 16.5 ± 3.5 (p > 0.05; c vs. a, b)Antioxidant action vs. control (plasma treated with H2O2/Fe) | 30.4 ± 10.9 (p < 0.05; c vs. a; p > 0.05; c vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 355.7 ± 13.7 (p > 0.05; c vs. a; p < 0.05; c vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | No effect vs. control |
Compound 1 (d) | 53.8 ± 12.7 (p < 0.05; d vs. a, b, c, e, f; p > 0.05; d vs. g) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 21.4 ± 8.7 (p > 0.05; d vs. c; p < 0.05; d vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 375.1 ± 11.6 ( p > 0.05; d vs. a, c; p < 0.05; d vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 11.1 ± 2.90 (p < 0.05; d vs. c) Anticoagulant action vs. control |
Compound 2 (e) | 25.0 ± 6.9 (p < 0.05; e vs. c; p > 0.05; e vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 18.5 ± 6.0 (p > 0.05; e vs. c; p < 0.05; e vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 395.1 ± 20.2 (p > 0.05; e vs. a, c; p < 0.05; e vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 18.3 ± 3.6 (p < 0.05; e vs. c) Anticoagulant action vs. control |
Compound 3 (f) | 27.7 ± 7.8 (p < 0.05; f vs. c; (p > 0.05; f vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 24.0 ± 7.0 (p > 0.05; f vs. c; p < 0.05; f vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 375.4 ± 16.4 (p > 0.05; f vs. a, c; p < 0.05; f vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 18.9 ± 5.4 (p < 0.05; f vs. c) Anticoagulant action vs. control |
Compound 4 (g) | 43.3 ± 10.5 (p < 0.05; g vs. a, b, c) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 7.8 ± 10.5 (p > 0.05; g vs. a, c; p < 0.05; g vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 443.6 ± 17.6 (p < 0.05; g vs. a, b, c. d, f) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 24.9 ± 6.8 (p < 0.05; g vs. b, c, d) Anticoagulant action vs. control |
Kaempferol (h) | 23.9 ± 4.9 (p > 0.05; h vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 20.8 ± 3.9 (p < 0.05; h vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 356.7 ± 14.2 (p > 0.05; h vs. a; p < 0.05; h vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | No effect vs. control |
Compound 5 (i) | 29.0 ± 5.0 (p > 0.05; i vs. a, b, h) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 24.5 ± 4.8 (p > 0.05; i vs. h; p < 0.05; i vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 358.8 ± 14.8 (p > 0.05; i vs. a, h; p < 0.05; i vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 23.3 ± 6.7 (p < 0.05; i vs. b, h) Anticoagulant action vs. control |
Compound 6 (j) | 29.8 ± 4.9 (p < 0.05; j vs. h; p > 0.05; j vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 18.2 ± 3.7 (p > 0.05; j vs. h; p < 0.05; j vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 390.3 ±14.2 (p > 0.05; j vs. a, h; p < 0.05; j vs. b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 22.2 ± 5.9 (p < 0.05; j vs. b, h) Anticoagulant action vs. control |
Compound 7 (k) | 57.8 ± 14.9 (p < 0.05; k vs. a, b, h, i, j) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 18.1 ± 3.9 (p > 0.05; k vs. h; p < 0.05; k vs. a, b) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 396.1 ± 14.3 (p > 0.05; k vs. a, b, h) Antioxidant action vs. control (plasma treated with H2O2/Fe) | 23.2 ± 7.2 (p < 0.05; k vs. b, h) Anticoagulant action vs. control |
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Żuchowski, J.; Rolnik, A.; Adach, W.; Stochmal, A.; Olas, B. Modulation of Oxidative Stress and Hemostasis by Flavonoids from Lentil Aerial Parts. Molecules 2021, 26, 497. https://doi.org/10.3390/molecules26020497
Żuchowski J, Rolnik A, Adach W, Stochmal A, Olas B. Modulation of Oxidative Stress and Hemostasis by Flavonoids from Lentil Aerial Parts. Molecules. 2021; 26(2):497. https://doi.org/10.3390/molecules26020497
Chicago/Turabian StyleŻuchowski, Jerzy, Agata Rolnik, Weronika Adach, Anna Stochmal, and Beata Olas. 2021. "Modulation of Oxidative Stress and Hemostasis by Flavonoids from Lentil Aerial Parts" Molecules 26, no. 2: 497. https://doi.org/10.3390/molecules26020497