A kind of antioxidant active peptide and its application
An active peptide and anti-oxidation technology, which is applied in applications, medical preparations containing active ingredients, peptides, etc., can solve the problems of unfavorable active peptide efficacy, low bioavailability, and poor absorption of active peptides, and achieve high oral absorption efficiency , strong proliferative ability, and high antioxidant activity
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experiment example 1
[0038] Determination of Antioxidative Ability of 12 Peptides by Hydroxyl Free Radical Scavenging Method
[0039] Prepare 1 mL of 1 mg / mL peptide solution, add 2 mL of 1.8 mM ferrous sulfate, 1.5 mL of 1.8 mM salicylic acid-ethanol solution, and 1 mL of 8.8 mM hydrogen peroxide solution in sequence, and incubate in a 37°C water bath for 30 min.
[0040] Prepare 1mL 1mg / mL walnut meal hydrolyzate solution, add 2mL 1.8mM ferrous sulfate, 1.5mL 1.8mM salicylic acid-ethanol solution, 1mL 8.8mM hydrogen peroxide solution, and incubate in a 37°C water bath for 30min.
[0041] Absorbance A1 was measured at 510 nm. Replace the sample with distilled water and detect according to the above method, and the measured absorbance is used as the control A0. Calculate the hydroxyl radical scavenging rate according to the following formula:
[0042] Hydroxyl radical scavenging rate (%)=[(A 0 -A 1 ) / A 0 ]×100%
[0043] The hydroxyl radical scavenging rate of 12 polypeptides is as follows: ...
experiment example 2
[0045] CCK-8 method to evaluate the effect of 12 peptides on the proliferation of oxidative damage to nerve cells
[0046] Construction of SH-SY5Y cell model with oxidative damage: SH-SY5Y cells in the logarithmic growth phase were seeded in 96-well cell culture plates, cultured overnight, and 200 μmol / L H 2 o 2 The solution was incubated with the cells for 24 hours, and the control group was set without any treatment. 10 μL of CCK-8 solution was added to each well, and the culture was terminated after 4 hours, and the absorbance at 450 nm was detected with a microplate reader. The measured cell survival rate of the model group was 73.1% of that of the control group, indicating that the oxidative damage model was successfully constructed.
[0047] Evaluation of proliferation-promoting ability: SH-SY5Y cells in the logarithmic growth phase were inoculated in 96-well cell culture plates, cultured overnight, and 200 μmol / L H 2 o 2 The solution was incubated with cells for 24h...
experiment example 3
[0049] Oral Absorption Efficiency Determination of 12 Peptides
[0050] Caco-2 monolayer cell model construction: Caco-2 cell monolayer model was selected to study the transport of representative peptides. Select Caco-2 cells in good growth state for digestion, and use 1×10 5 piece / cm 2 Inoculated on the polycarbonate membrane of the Millicell chamber, inserted the Millicell chamber into a 24-well plate, and placed it in an incubator at 37°C, 5% CO 2 cultivated in the environment. The medium was changed 24 hours after inoculation, and then every 2 days. After culturing for 21 days, the transmembrane resistance value TEER was measured. TEER values are greater than 300Ω·m 2 , indicating that the Caco-2 monolayer cell model was successfully constructed.
[0051]Caco-2 cell transport experiment: First, 15 polypeptides were fluorescently labeled with 5-FAM, and a 3 mg / mL solution was prepared as a dosing solution. Then, the liquid on the top and bottom sides of the Millice...
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