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Protective Effect of Whey Protein Hydrolysates on Hydrogen Peroxide-induced Oxidative

Author: LingYuFang
Tutor: LuRongRong
School: Jiangnan University
Course: Of Food Science
Keywords: Whey protein Of Hydrolysates Radicals Antioxidant Apoptosis PC12 cells
CLC: R151
Type: Master's thesis
Year: 2011
Downloads: 85
Quote: 1
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Abstract


Natural antioxidant peptide because of its natural and safety in the field of anti-oxidation, high-profile, and therefore has very broad prospects for development and application of protein source of antioxidant peptides. Whey protein as a high-quality protein resources, has many researchers concerned about, and to the development of biologically active peptides. The study found that whey protein can be extracted with the antioxidant activity of biological peptide, however, by far the majority of studies mainly focused on whey protein hydrolyzate process optimization and preliminary purification process. This research aims to reveal the the Ruyuan nature antioxidant peptide biological effects of oxidative damage to cells in the body, the perfect antioxidant peptides stress mechanism to guide the direction of application of the antioxidant peptide, but also the study of the mechanism of action of the functional material provide the methodology learning has theoretical significance and expand the scientific significance. Firstly, whey protein hydrolysis by enzymes (pepsin - trypsin), the crude enzyme preparation after DA201-C Model macroporous adsorption resin adsorption elution off a lot of salt, the relative molecular weight of the product (WPHs) distributed in 61.51% of the total area of ??the peak area of ??the 369 1 980 Da, 1000 Da fragment; right WPHs the amino acid composition analysis shows WPHs rich in essential amino acids (43.72%) and a hydrophobic amino acid (40.75 %); In addition, WPHs vitro antioxidant activity, DPPH radical scavenging superoxide anion radical IC50, respectively, to 10.56 mg / mL, 5.76 mg / mL, with a concentration of 10 mg / mL, restore the ability A700nm 0.577 . PC12 cells were seeded to plates incubated overnight, after the different concentrations WPHs pre-incubation 2 h, WPHs 100μmol/LH 2 O 2 common treatment for 24 h. Cell viability (MTT assay) results show 100 4 00μg/mL concentration WPHs H 2 O 2 induced damage in PC12 cells can survive increased by 20% to 30% of the rate; WPHs to promote the growth of the cells themselves have, the concentration of 100 4 of 00μg/mL WPHs alone 24 h, significant growth-promoting effect of 1%, improve the survival rate of PC12 cells < sup> 1 9%; 100 2 00μg/mL WPHs improvement in the form of hydrogen peroxide induced damage in PC12 cells, can significantly reduce shrink and the number of suspension cells. The thesis WPHs inhibition mechanism of hydrogen peroxide-induced oxidative damage. Enzyme kit to detect the activity of intracellular antioxidant enzyme system, WPHs PC12 cell antioxidant capacity and antioxidant enzyme system improved; detection of apoptosis using the nuclear staining, results showed that 100 2 < / sup> 00μg/mL WPHs inhibit H 2 O 2 PC12 cells induced apoptosis, respectively, the apoptosis rate declined 8.3% 1 9%; cells using flow cytometry Ca 2 concentration and mitochondrial transmembrane potential, found the 200μg/mL WPHs can stabilize the intracellular Ca 2 concentration prevent mitochondrial membrane potential and change; PC12 cells Caspase-3 enzyme activity detected using Caspase-3 activity kit, the discovery WPHs can reduce hydrogen peroxide induced induced Caspase-3 activity increased degree of; and apoptosis proteins The expression levels detected by Western blot, The results show WPHs be able to improve the anti-apoptotic protein in PC12 cells the expression of Bcl-2, inhibit the expression of Bax protein and reduce the degree of degradation of PARP. The results of this study show that, WPHs containing large amounts of small peptides, hydrogen peroxide oxidation damage to cells has good protection, mainly by way of maintain the normal operation of the cell signaling pathways, inhibiting cell apoptosis caused by the peroxide injury. The enzymatic hydrolysis was resistant to digestion, an antioxidant, suitable as a health food or food additive.

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CLC: > Medicine, health > Preventive Medicine,Health > Nutrition, hygiene,food hygiene > Nutrition
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