The Pro-Gly or Hyp-Gly Containing Peptides from Absorbates of Fish Skin Collagen Hydrolysates Inhibit Platelet Aggregation and Target P2Y12 Receptor by Molecular Docking
Abstract
:1. Introduction
2. Materials and Methods
2.1. Material and Chemicals
2.2. Preparation of Collagen Peptides (CPs)
2.3. Simulated Gastrointestinal Digestion
2.4. Uptake and Transport by Caco-2 Cells
2.5. Separation of the AH Absorbate (AHA) and PH Absorbate (PHA)
2.6. Peptide Sequencing Using LC-MS/MS
2.7. Antiplatelet Activity Assay In Vitro
2.8. Animals, Diets, and Treatments
2.8.1. Feeding Conditions and Animal
2.8.2. Ferric Chloride-Induced Arterial Thrombosis Model
2.8.3. Bleeding Assay and Determination of Thymus Index and Spleen Index
2.8.4. Determination of Thymus Index and Spleen Index
2.9. Molecular Docking
2.10. Statistics
3. Results
3.1. Effect of Alcalase-Hydrolysates (AH) and Protamex®-hydrolysates (PH) on Inhibiting Platelet Aggregation Induced by Collagen, Thrombin, and ADP In Vitro
3.2. Separation and Identification of Antiplatelet Peptides from Collagen Hydrolysates after Simulated Gastrointestinal Digestion and Intestinal Absorption
3.3. Effect of Identified Peptides on Inhibiting Platelet Aggregation Induced by Collagen, Thrombin, and ADP In Vitro
3.4. Tetrapeptide OGSA Inhibited Thrombus Formation In Vivo
3.5. Molecular Docking of Antiplatelet Peptides with P2Y12 Receptor
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
AHA | Absorbate of Alcalase hydrolysate |
PHA | Absorbate of Protamex® hydrolysate |
APTT | Activated partial thromboplastin time |
ADP | Adenosine diphosphate |
bw | Body weight |
CCVd | Cardiocerebrovascular diseases |
Clo | Clopidogrel |
CPs | Collagen peptides |
HP | High-dose peptides group |
LP | Low-dose peptides group |
N | Normal group |
PRP | Platelet-rich plasma |
PPP | Poor platelet plasma |
PT | Prothrombin time |
SI | Spleen index |
TT | Thrombin time |
TI | Thymus index |
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Sample | Peptide a | Inhibition Rate of Platelet Aggregation % | ||
---|---|---|---|---|
Collagen Induced | Thrombin Induced | ADP b Induced | ||
Control | RGD | 17.1 ± 3.1 | 22.7 ± 2.7 | 79.7 ± 2.8 |
Aspirin d | 55.0 ± 7.2 | 6.1 ± 2.5 | 58.4 ± 5.5 | |
A1 c | EGPAGPA | 16.4 ± 2.2 | 53.9 ± 2.1 | 90.7 ± 4.4 |
GTOGT | 5.1 ± 27.7 | −2.7 ± 7.1 | 88.0 ± 4.4 | |
PGPK | −18.7 ± 19.7 | −2.4 ± 5.9 | 69.3 ± 0.5 | |
PGKP | −56.0 ± 10.7 | 0.1 ± 4.6 | 39.8 ± 1.9 | |
PGPQ | −51.2 ± 26.5 | 0.9 ± 1.0 | 64.5 ± 16.2 | |
PGQP | 65.6 ± 14.1 | −2.4 ± 2.9 | 56.2 ± 1.1 | |
OGSA | −0.1 ± 24.2 | 15.5 ± 4.4 | 96.4 ± 2.0 | |
OG | 6.5 ± 0.7 | −6.5 ± 2.2 | 56.6 ± 1.9 | |
A4 c | VVGOKG | −9.0 ± 0.9 | 11.5 ± 2.0 | 96.0 ± 1.7 |
OGOMG | 12.4 ± 19.4 | 0.3 ± 3.4 | 73.6 ± 7.4 | |
P3 c | PGHH | 21.3 ± 8.7 | 19.6 ± 4.9 | 83.7 ± 3.5 |
No. | Sequence a | IC50/mM b | PLC β2 c | PLC β3 c | P2Y12 c |
---|---|---|---|---|---|
1 | OGE | 0.62 | −5.05 | −5.24 | −6.62 |
2 | OGSA | 0.63 | −5.92 | −3.88 | −5.73 |
3 | PGE | 0.93 | −5.22 | −4.72 | −6.09 |
4 | PGEOG | 0.97 | −4.57 | −4.44 | −5.71 |
5 | PGHH | 1.4 | −4.78 | −4.33 | −4.21 |
6 | OGOMG | 1.49 | −4.55 | −4.25 | −3.93 |
7 | PGPK | 2.79 | −6.24 | −3.83 | −3.27 |
8 | OG | 1.68 | −6.11 | −4.68 | −4.68 |
9 | PGKP | 2.57 | −4.6 | −3.68 | −2.92 |
10 | VGPOGPA | 2.73 | −4.84 | −3.11 | −1.40 |
11 | GTOGT | 3.71 | −2.00 | −2.04 | −1.46 |
12 | PGPQ | 3.56 | −6.47 | −5.66 | −4.53 |
13 | PGQP | 2.31 | −5.29 | −5.21 | −3.23 |
Pearson correlation coefficient | 0.220 | 0.367 | 0.802 |
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Tian, Q.; Li, S.-M.; Li, B. The Pro-Gly or Hyp-Gly Containing Peptides from Absorbates of Fish Skin Collagen Hydrolysates Inhibit Platelet Aggregation and Target P2Y12 Receptor by Molecular Docking. Foods 2021, 10, 1553. https://doi.org/10.3390/foods10071553
Tian Q, Li S-M, Li B. The Pro-Gly or Hyp-Gly Containing Peptides from Absorbates of Fish Skin Collagen Hydrolysates Inhibit Platelet Aggregation and Target P2Y12 Receptor by Molecular Docking. Foods. 2021; 10(7):1553. https://doi.org/10.3390/foods10071553
Chicago/Turabian StyleTian, Qi, Shi-Ming Li, and Bo Li. 2021. "The Pro-Gly or Hyp-Gly Containing Peptides from Absorbates of Fish Skin Collagen Hydrolysates Inhibit Platelet Aggregation and Target P2Y12 Receptor by Molecular Docking" Foods 10, no. 7: 1553. https://doi.org/10.3390/foods10071553