Viburnum opulus Fruit Phenolic Compounds as Cytoprotective Agents Able to Decrease Free Fatty Acids and Glucose Uptake by Caco-2 Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals and Reagents
2.2. Preparation of Viburnum Opulus Extracts
2.3. Determination of Total Phenolics Content and Phenolic Compounds Profile
2.4. Antioxidant Capacity Assays
2.5. Cell Culture and Exposure Conditions
2.6. Cell Viability
2.7. Detection of Intracellular Reactive Oxygen Species (ROS) Generation
2.8. Measurement of ATP Production
2.9. Measurement of Mitochondrial Membrane Potential (MMP)
2.10. Phosphatidylserine Externalization, Membrane Permeabilization and Apoptosis Induction
2.11. Glucose Uptake
2.12. Nile Red Staining
2.13. Fatty Acid Uptake
2.14. DNA Damage and Repair
2.15. Gene Expression Analysis
2.16. Statistical Analysis
3. Results and Discussion
3.1. Phenolic Compounds Profile and Content
3.2. Antioxidant Capacity of V. opulus Extracts
3.3. Assessment of Viburnum Opulus Phenolics on Cellular Viability
3.4. Viburnum opulus as a Chemopreventive Agent
3.5. Effect of V. opulus Extracts on Intracellular ATP Level, Intracellular Oxidative Stress and Apoptosis Induction
3.6. Viburnum opulus Phenolics as Modulators of Fatty Acids Uptake
3.7. Viburnum opulus Phenolic Compounds as Modulators of Glucose Uptake
4. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Hydroxybenzoic Acid (HBA) | Rt [min] | λmax [nm] | FJ | MAE | AE | PRF |
---|---|---|---|---|---|---|
mg/g of Juice | mg/g of Extract | |||||
Gallic acid | 1.92 | 282.7 | n.d. | 0.21 ± 0.00 a | 0.40 ± 0.00 b | n.d. |
Flavanols | 1.21 ± 0.01 | 22.84 ± 0.05 | 15.46 ± 0.22 | 100.74 ± 0.10 | ||
Procyanidin B1 | 4.50 | 278.7 | 0.31 ± 0.00 b | 3.28 ± 0.03 c | 2.46 ± 0.01 a | 29.76 ± 0.05 d |
(+)-Catechin | 5.19 | 278.7 | 0.68 ± 0.01 b | 9.71 ± 0.01 c | 5.70 ± 0.00 a | 53.18 ± 0.03 d |
Procyanidin B2 | 5.98 | 278.7 | 0.22 ± 0.00 a | 5.44 ± 0.02 c | 3.77 ± 0.03 b | 17.79 ± 0.02 d |
(−)-Epicatechin | 6.85 | 277.7 | n.d. | 4.42 ± 0.02 b | 3.53 ± 0.18 a | n.d. |
Hydroxycinnamic Acids (HCA) | 7.97 ± 0.02 | 96.37 ± 0.14 | 84.27 ± 0.26 | 645.85 ± 2.00 | ||
Neochlorogenic acid | 3.94 | 323.7 | n.d. | 0.16 ± 0.00 b | 0.15 ± 0.00 a | 0,36 ± 0.00 c |
Chlorogenic acid | 5.55 | 325.7 | 7.97 ± 0.02 a | 96.22 ± 0.14 c | 84.31 ± 0.26 b | 645.49 ± 2.00 |
Flavonols | 0.01 ± 0.00 | 0.54 ± 0.00 | 0.48 ± 0.00 | 2.35 ± 0.00 | ||
Rutin | 9.66 | 353.7 | 0.01 ± 0.00 b | n.d. | 0.12 ± 0.00 a | 1.09 ± 0.00 c |
Isorhamnetin | 9.93 | 353.7 | 0.002 ± 0.000 a | n.d. | n.d. | 0.12 ± 0.00 b |
Isorhamnetin 3-O-rutinoside | 11.20 | 353.7 | 0.01 ± 0.00 a | 0.26 ± 0.00c | 0.18 ± 0.00 b | 0.84 ± 0.00 d |
Quercetin | 11.55 | 352.7 | n.d. | 0.28 ± 0.00 b | 0.17 ± 0.00 a | 0.30 ± 0.00 c |
Anthocyanins | 1.12 ± 0.00 | 2.07 ± 0.01 | 1.45 ± 0.00 | 78.06 ± 0.68 | ||
Cyanidin-3-O-sambubioside | 6.83 | 516.7 | 0.85 ± 0.00 | 0.74 ± 0.00 | 0.56 ± 0.00 | 55.78 ± 0.69 |
Cyanidin-3-O-glucoside | 7.03 | 515.7 | 0.20 ± 0.01 | 0.86 ± 0.00 | 0.58 ± 0.00 | 17.08 ± 0.01 |
Cyanidin-3-O-rutinoside | 7.36 | 518.7 | 0.06 ± 0.00 | 0.47 ± 0.00 | 0.31 ± 0.00 | 5.21 ± 0.01 |
Phenolics Total | 10.32 ± 0.03 | 121.83 ± 0.19 | 101.67 ± 0.41 | 827.00 ± 1.68 |
Sample | Total Phenolics | Antioxidant Capacity | |
---|---|---|---|
ABTS | ORAC | ||
mg GAE/g | µmol TE/g | ||
Methanol-Acetone Extract (MAE) | 143.77 ± 4.0 b | 1230.81 ± 26.1 b | 2388.58 ± 150.5c |
Acetone Extract (AE) | 137.80 ± 5.6 b | 1227.73 ± 32.0 b | 2092.65 ± 48.3 b |
Phenolic Rich Fraction (PRF) | 361.52 ± 13.3 c | 2619.59 ± 123.1c | 7810.29 ± 342.3d |
Fresh Juice (FJ) | 5.98 ± 0.3 a | 38.36 ± 2.7 a | 103.50 ± 6.4 a |
V. opulus Preparation | IC0 [µg/mL] | IC50 [µg/mL] | Cells Viability [%] 1 | |
t-BOOH 500 μM | 79.50 ± 3.85 | |||
FJ | 75.0 | 450.0 | 95.64 ± 8.78 | |
PRF | 50.0 | 250.0 | 96.12 ± 10.12 | |
MAE | 50.0 | 275.0 | 91.70 ± 8.31 | |
AE | 100 | 400.0 | 90.50 ± 3.02 |
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Zakłos-Szyda, M.; Pawlik, N.; Polka, D.; Nowak, A.; Koziołkiewicz, M.; Podsędek, A. Viburnum opulus Fruit Phenolic Compounds as Cytoprotective Agents Able to Decrease Free Fatty Acids and Glucose Uptake by Caco-2 Cells. Antioxidants 2019, 8, 262. https://doi.org/10.3390/antiox8080262
Zakłos-Szyda M, Pawlik N, Polka D, Nowak A, Koziołkiewicz M, Podsędek A. Viburnum opulus Fruit Phenolic Compounds as Cytoprotective Agents Able to Decrease Free Fatty Acids and Glucose Uptake by Caco-2 Cells. Antioxidants. 2019; 8(8):262. https://doi.org/10.3390/antiox8080262
Chicago/Turabian StyleZakłos-Szyda, Małgorzata, Nina Pawlik, Dominika Polka, Adriana Nowak, Maria Koziołkiewicz, and Anna Podsędek. 2019. "Viburnum opulus Fruit Phenolic Compounds as Cytoprotective Agents Able to Decrease Free Fatty Acids and Glucose Uptake by Caco-2 Cells" Antioxidants 8, no. 8: 262. https://doi.org/10.3390/antiox8080262