Antioxidant and Antiapoptotic Properties of Oat Bran Protein Hydrolysates in AAPH-Induced Oxidative Stress Hepatocarcinoma HepG2
Notice bibliographique
Résumé
The onset of many chronic diseases such as cancer, carcinogenesis, cardiovascular disease, and several other health problems, involves oxidative stress damage to biomolecules such as DNA, lipid, and proteins.There have been many preventative and therapeutic methods proposed to increase cellular defense mechanisms against oxidative damage such as using bioactive peptide with antioxidant activities from food sources.This study found that AAPH-induced oxidative stress in human hepatocarcinoma HepG2 cells caused changes in cellular morphological features and functionalities such as cell viability, activity of antioxidative enzymes (catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD)), production of reactive oxygen species (ROS), and rate of apoptosis.The use of oat bran protein hydrolysates restored some of the properties to normal values (i.e., control cells).In this work, oat brans were treated with two carbohydrases (Cellulase ® (Cell) and viscozyme ® L (Visc) which cleave cell wall polysaccharides and produce oat bran protein isolates (OBPIs) with protein contents.OBPI was subsequently treated with proteases Alcalase (Alca), Flavourzyme ® (Flav), Papain (Papa) and Protamex (Prot) to produce four oat bran protein hydrolysates (OBPHs) from each of oat bran protein isolate (OBPIs).Antioxidative effects of both the two OBPI and four OBPH samples were studied in chemistry and HepG2 cell cultures assays.In the chemistry assays the antioxidative activities were determined using oxygen radical absorbance capacity (ORAC), 2,2'-azino-bis (3ethylbenzothiazoline-6-sulphonic acid) or ABTS •+ , radical, hydroxyl ( • OH), and superoxide anion (O2 •-) radical scavenging assays.Data from oxygen radical absorbance capacity (ORAC) and 2,2'azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS •+ ) assays were expressed as Trolox (a water-soluble vitamin E analogue) equivalent per gram (TE/g) of the sample followed the same pattern.The highest ORAC activities were observed in Cell/Papa (417.0 ± 5.1 TE/g), Visc/Flav (406.0 ± 8.8 TE/g) and Visc/Papa (397.0 ± 13.9 TE/g) while in the ABTS •+ , highest scavenging samples were Visc/Flav and Visc/Papa (863.80 ± 20.3 and 814.8 ± 17.2 TE/g respectively).In the hydroxyl radical scavenging assay, Cellulase and Viscozyme OBPIs significantly prevented the formation of • OH, radicals 38.5 ± 0.8 and 36.0 ± 0.4%, respectively compared to all hydrolysates.iii Superoxide anion radical scavenging activity was the highest in samples Visc/Alca and Cell/Prot both at 37.3 ± 0.9%.SDS-PAGE results indicated that globulin (58 kDa) was the main protein in OPBIs and those hydrolysates did not contain protein or fragments above 15 kDa.Cytotoxicity of OBPIs and OBPHs, their ability to reduce intracellular reactive oxygen species (ROS), and regulate the activity of three main antioxidant enzymes were evaluated in the AAPH-induced oxidative stress of HepG2 cells.Cytotoxic effect OBPH was evaluated using Visc/Flav ND at 50 -400 µg/mL.No cytotoxicity was observed in MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay.In the cytoprotection assay, Viscozyme and all its OBPHs derivatives were able to increase cell viability significantly compared to stressed control HepG2 cells.However, Cellulase OBPI and its OBPHs derivatives enhanced cell HepG2 cell death, except for Cell/Flav OBPH which had no effect.OBPHs from Viscozyme treated brans and Cell/Flav OBPH were then chosen for investigation of their effects on antioxidant enzymes.In the Catalase (CAT) assay, all samples increased enzyme activity (p < 0.05) in AAPH-stressed cells, with Visc/Papa being the highest at 352.0%.In superoxide dismutase (SOD), all samples increased activity level above positive control (POS) with Visc/Flav performing significantly better than the other samples.Visc/Flav not only restored glutathione peroxidase (GPx) activity to that of negative control (NEG) (i.e., unstressed cells), it also reduced the most intracellular reactive oxygen species (ROS).Total glutathione (GSH) increased up to 6-fold in the presence of OBPH in stressed HepG2 cells.In the Caspase-3 assay for anti-apoptosis properties, samples reduced caspase-3 significantly compared to AAPH-induced oxidative stress in HepG2 cells.OBPH protected HepG2 cells from oxidative stress due to a combination of factors including radical scavenging activity, upregulation of antioxidant enzymes, and inhibition of the activity of the apoptotic enzyme caspase-3.v
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Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,001 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».