Anti-Inflammatory Activity of Haskap Cultivars is Polyphenols-Dependent
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Material
2.2. Chemicals
2.3. Extraction
2.3.1. Total Phenolic Content
2.3.2. Total Flavonoid Content
2.3.3. Total Anthocyanin Content
2.3.4. Total Proanthocyanidin Content
2.3.5. LC-MS/MS Analysis of Specific Polyphenols
2.3.6. Sugars and Organic Acid Analyses
2.4. Cell Culture
2.4.1. Measurement of Cell Viability
2.4.2. Measurement of Nitric Oxide
2.4.3. Measurement of COX-2 Activity
2.4.4. Measurement of IL-6 and TNF-α
2.4.5. Measurement of PGE2
2.5. Statistical Analysis
3. Results and Discussion
3.1. Qualitative Phenolic Composition
Growing Location | Cultivar | Total Phenolics (mg GAE/100 g FW) | Total Flavonoids (mg QE/100 g FW) | Total Proanthocyanidins (mg CE/100 g FW) | Total Anthocyanins (mg CGE/100 g FW) |
---|---|---|---|---|---|
LaHave farm | BL | 755.9 ± 9.4 d,e,f | 1156.6 ± 121.7 b,c | 13.2 ± 1.0 c | 163.0 ± 10.1 c |
BR | 1154.1 ± 59.7 a | 1582.8 ± 140.5 a | 16.3 ± 0.9 c | 314.0 ± 2.7 a | |
TN | 952.9 ± 18.7 b,c | 1260.3 ± 69.0 b,c | 16.0 ± 1.0 c | 234.4 ± 2.6 b | |
IG | 884.3 ± 25.0 c,d | 1327.0 ± 12.1 b,c | 14.4 ± 0.5 c | 246.9 ± 13.7 b | |
Kentville | LC-12 | 849.2 ± 28.3 c,d | 1035.5 ± 86.7 b,c | 37.2 ± 0.9 b | 164.9 ± 5.4 c |
LC-13 | 796.8 ± 5.2 d | 997.0 ± 32.0 b,c | 41.0 ± 3.6 b | 142.5 ± 9.1 c | |
LC-16 | 664.9 ± 51.4 e,f | 900.7 ± 16.6 c,d | 52.3 ± 2.3 a | 70.2 ± 2.2 d | |
LC-23 | 658.1 ± 0.4 e,f | 956.3 ± 11.0 b,c | 34.2 ± 0.8 b | 120.2 ± 1.5 c,d | |
LC-47 | 634.4 ± 29.9 f | 916.5 ± 65.2 c,d | 47.0 ± 0.4 a,b | 133.4 ± 1.2 c | |
Saskatchewan | SAS-IG | 790.3 ± 68.4 d | 1128.5 ± 54.2 c,d | 19.6 ± 2.0 c | 246.3 ± 2.8 b |
SAS-TN | 1015.3 ± 78.2 a,b | 1428.4 ± 35.1 a,b | 38.5 ± 0.9 b | 303.2 ± 5.5 a |
3.1.1. Total Flavonoid Content
3.1.2. Total Anthocyanin Content
3.1.3. Total Proanthocyanidin Content
3.1.4. LC-MS/MS Composition of Haskap Berry Extract
LaHave Farm | Kentville | Saskatchewan | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
Compounds | BR | BL | TN | IG | LC-12 | LC-13 | LC-16 | LC-23 | LC-47 | SAS-IG | SAS-TN |
Phenolic acids | |||||||||||
Chlorogenic acid | 25.6 ± 3.6 | 23.1 ± 2.4 | 26.0 ± 2.4 | 22.4 ± 2.5 | 29.7 ± 0.6 | 32.7 ± 3.2 | 23.0 ± 0.2 | 23.9 ± 1.3 | 28.6 ± 0.1 | 20.7 ± 1.1 | 33.8 ± 0.7 |
Caffeic acid | 0.2 ± 0.0 | 0.1 ± 0.0 | 0.2 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | ND | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 |
Total | 25.8 | 23.2 | 26.2 | 22.5 | 29.8 | 32.8 | 23.0 | 24.0 | 28.7 | 20.8 | 33.9 |
Flavan-3-ols | |||||||||||
EGC | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.6 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 |
Catechin | 2.5 ± 0.2 | 1.7 ± 0.3 | 3.5 ± 0.2 | 2.9 ± 0.3 | 2.1 ± 0.1 | 2.1 ± 0.3 | 3.9 ± 0.1 | 5.4 ± 0.0 | 2.1 ± 0.1 | 2.2 ± 0.1 | 3.4 ± 0.2 |
Epicatechin | 1.2 ± 0.1 | 1.7 ± 0.4 | 0.7 ± 0.1 | 1.5 ± 0.1 | 4.5 ± 0.1 | 5.8 ± 0.5 | 7.1 ± 0.1 | 6.2 ± 0.1 | 1.5 ± 0.1 | 0.9 ± 0.0 | 0.9 ± 0.0 |
EGCG | 0.1 ± 0.0 | 0.2 ± 0.0 | 0.2 ± 0.0 | 0.2 ± 0.0 | 0.1 ± 0.0 | 0.3 ± 0.0 | 0.1 ± 0.0 | 0.3 ± 0.0 | 0.2 ± 0.0 | 0.2 ± 0.0 | 0.2 ± 0.0 |
Total | 3.9 | 3.7 | 4.5 | 5.1 | 6.8 | 8.3 | 11.2 | 12.0 | 4.0 | 3.4 | 4.6 |
Flavonols | |||||||||||
Q. galactoside | ND | 0.1 ± 0.0 | ND | ND | 0.1 ± 0.0 | 0.1 ± 0.0 | ND | ND | ND | ND | ND |
Q. glucoside | 3.6 ± 0.2 | 4.2 ± 0.8 | 1.4 ± 0.2 | 2.8 ± 0.2 | 7.3 ± 0.8 | 4.4 ± 0.6 | 3.7 ± 0.1 | 3.8 ± 0.1 | 3.7 ± 0.1 | 2.7 ± 0.1 | 4.0 ± 0.2 |
Q. arabinoside | 2.9 ± 0.3 | 2.0 ± 0.3 | 1.4 ± 0.2 | 1.1 ± 0.1 | 11.9 ± 0.8 | 9.2 ± 0.8 | 11.4 ± 0.3 | 7.3 ± 0.4 | 10.0 ± 0.0 | 1.0 ± 0.0 | 2.9 ± 0.1 |
Q. rhamnoside | 0.1 ± 0.0 | 0.2 ± 0.0 | 0.0 ± 0.0 | 0.1 ± 0.0 | 2.2 ± 0.2 | 1.2 ± 0.2 | 0.4 ± 0.0 | 0.3 ± 0.0 | 0.4 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 |
Q | 0.2 ± 0.0 | 0.3 ± 0.0 | 0.2 ± 0.0 | 0.2 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.2 ± 0.0 | 0.1 ± 0.0 |
Q. rutinoside | 24.3 ± 1.2 | 16.7 ± 6.0 | 19.9 ± 3.0 | 21.6 ± 0.6 | 8.5 ± 0.3 | 6.2 ± 0.2 | 7.7 ± 0.6 | 10.9 ± 1.1 | 6.7 ± 0.1 | 19.5 ± 1.3 | 20.4 ± 1.3 |
Total | 31.1 | 23.5 | 22.9 | 25.8 | 30.1 | 21.2 | 23.3 | 22.4 | 20.9 | 23.5 | 27.5 |
Dihydrochalcones | |||||||||||
Phloridzin | 0.4 ± 0.0 | 0.3 ± 0.0 | 0.2 ± 0.0 | 0.3 ± 0.0 | 0.25 ± 0.0 | 0.2 ± 0.0 | 0.1 ± 0.0 | 0.2 ± 0.0 | 0.1 ± 0.0 | 0.3 ± 0.0 | 0.1 ± 0.0 |
Anthocyanins | |||||||||||
C-3-gluc | 170.0 ± 10.1 | 140.8 ± 8.3 | 104.7 ± 10.3 | 143.9 ± 1.5 | 147.2 ± 5.6 | 107.7 ± 2.1 | 67.7 ± 2.7 | 103.4 ± 3.1 | 107.2 ± 0.1 | 138.8 ± 8.2 | 164.3 ± 6.2 |
D-3-glu | 0.4 ± 0.0 | 0.4 ± 0.0 | 0.3 ± 0.0 | 0.4 ± 0.0 | 0.1 ± 0.0 | 0.2 ± 0.0 | 0.2 ± 0.0 | 0.2 ± 0.0 | 0.2 ± 0.0 | 0.3 ± 0.0 | 0.4 ± 0.0 |
P-3-gluc | 13.7 ± 0.9 | 9.5 ± 0.5 | 6.7 ± 0.7 | 8.2 ± 0.3 | 9.9 ± 0.1 | 4.3 ± 0.2 | 3.9 ± 0.3 | 7.6 ± 0.3 | 4.9 ± 0.3 | 7.8 ± 0.3 | 14.7 ± 0.4 |
D-3-rutin | 0.2 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.4 ± 0.0 | 0.2 ± 0.0 | 0.1 ± 0.0 | 0.0 ± 0.0 | 0.1 ± 0.0 | 0.0 ± 0.0 | 0.1 ± 0.0 |
C-3-rutin | 39.2 ± 2.3 | 26.1 ± 7.5 | 64.9 ± 9.5 | 38.6 ± 2.5 | 19.5 ± 1.0 | 17.4 ± 0.6 | 11.8 ± 1.3 | 32.0 ± 0.2 | 22.7 ± 0.2 | 31.9 ± 1.3 | 34.6 ± 1.1 |
C-3-galact | 0.4 ± 0.0 | 0.2 ± 0.0 | 0.8 ± 0.1 | 0.0 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.1 ± 0.0 | 0.1 ± 0.0 | 0.4 ± 0.0 |
Total | 223.9 | 177.1 | 177.5 | 191.2 | 177.2 | 129.9 | 83.7 | 143.2 | 135.2 | 178.9 | 214.5 |
Total phenolics by LCMS | 285.1 a | 227.8 c,d | 231.29 c,d | 244.87 b,c | 244.25 b,c | 192.5 f | 209.12 c,d,e | 201.92 d,e | 188.9 f | 226.9 c,d | 280.6 a |
3.1.5. Sugar and Organic Acid Profile
3.2. Inhibition of Inflammatory Markers by Haskap Berry Extract
Parameters | COX-2 | TNF-α | IL-6 | PGE2 | NO |
---|---|---|---|---|---|
Phenolics | −0.781, 0.003 | −0.935, 0.000 | −0.896, 0.000 | −0.026, 0.936 | 0.324, 0.304 |
Flavonoids | −0.742, 0.006 | −0.781, 0.003 | −0.723, 0.008 | −0.186, 0.563 | 0.264, 0.406 |
Anthocyanins | −0.728, 0.007 | −0.831, 0.001 | −0.629, 0.028 | 0.005, 0.988 | 0.327, 0.300 |
Proanthocyanidins | −0.211, 0.511 | −0.290, 0.360 | −0.298, 0.348 | 0.459, 0.134 | 0.084, 0.796 |
3.3. Correlation between Phenolics and Inflammatory Parameters
4. Conclusions
Acknowledgements
Author Contributions
Appendix
Conflict of Interests
References
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Rupasinghe, H.P.V.; Boehm, M.M.A.; Sekhon-Loodu, S.; Parmar, I.; Bors, B.; Jamieson, A.R. Anti-Inflammatory Activity of Haskap Cultivars is Polyphenols-Dependent. Biomolecules 2015, 5, 1079-1098. https://doi.org/10.3390/biom5021079
Rupasinghe HPV, Boehm MMA, Sekhon-Loodu S, Parmar I, Bors B, Jamieson AR. Anti-Inflammatory Activity of Haskap Cultivars is Polyphenols-Dependent. Biomolecules. 2015; 5(2):1079-1098. https://doi.org/10.3390/biom5021079
Chicago/Turabian StyleRupasinghe, H. P. Vasantha, Mannfred M. A. Boehm, Satvir Sekhon-Loodu, Indu Parmar, Bob Bors, and Andrew R. Jamieson. 2015. "Anti-Inflammatory Activity of Haskap Cultivars is Polyphenols-Dependent" Biomolecules 5, no. 2: 1079-1098. https://doi.org/10.3390/biom5021079