Cellular Antioxidant Activity Research Articles

Cymensifin A: a promising pharmaceutical candidate to defeat lung cancer via cellular reactive oxygen species-mediated apoptosis

Background: The upgrade of natural products for cancer treatment is essential since current anticancer drugs still pose severe side effects. Cymensifin A (Cym A) isolated from an orchid Cymbidium ensifolium has shown its potential to induce the death of several cancer cells; however, its underlying molecular mechanisms are hitherto unknown.Methods: Here, we conducted a set of in vitro preliminary tests to assess the cytotoxic effects of Cym A on non-small-cell lung cancer (NSCLC) cells (A549, H23, H292, and H460). A flow cytometry system and Western blot analyses were employed to unveil molecular mechanisms underlying cancer cell apoptosis caused by Cym A.Results: Cym A at 25–50 μM caused the death of all NSCLC cells tested, and its cytotoxicity was comparable to cisplatin, a currently used anticancer drug. The compound induced apoptosis of all NSCLC cells in a dose-dependent manner (5–50 μM), proven by flow cytometry, but H460 cells showed more resistance compared to other cells tested. Cym A-treated H460 cells demonstrated increased reactive oxygen species (ROS) and downregulated antioxidants (catalase, superoxide dismutase, and thioredoxin). The compound also upregulated the tumor suppressor P53 and the pro-apoptotic protein BAX but downregulated pro-survival proteins (BCL-2 and MCL-1) and deactivated survival signals (AKT and ERK) in H460 cells. Cym A was proven to trigger cellular ROS formation, but P53 and BAX were 2-fold more activated by Cym A compared to those treated with hydrogen peroxide. Our findings also supported that Cym A exerted its roles in the downregulation of nuclear factor erythroid 2–related factor 2 (a regulator of cellular antioxidant activity) and the increased levels of cleaved poly (ADP-ribose) polymerase (PARP) and cleaved caspase 3/7 during apoptosis.Conclusion: We propose that Cym A induces lung cancer cell death via ROS-mediated apoptosis, while the modulation of cellular ROS/antioxidant activity, the upregulation of P53 and BAX, the downregulation or deactivation of BCL-2, MCL-1, AKT, and ERK, and the increased cleavage of PARP and caspase 3/7, were the elucidated underlying molecular mechanisms of this phytochemical. The compound can be a promising candidate for future anticancer drug development.

Identification and molecular mechanisms of novel antioxidant peptides from fermented broad bean paste: A combined in silico and in vitro study

This study aimed to investigate the antioxidative and cytoprotective activity of antioxidant peptides from fermented broad bean paste (FBBP) and explore their potential molecular mechanisms using a combined in silico and in vitro approach. Seven novel antioxidant peptides (VSRRFIYYL, SPAIPLP, PVPPPGG, KKDGYWWAKFK, LAWY, LGFMQF, and LPGCP) identified by integrated approaches of peptidomics and in silico bioinformatic analysis were synthesized, exhibiting strong antioxidant potential against in vitro radicals. Molecular docking results suggested that these peptides could form stable hydrogen bonds and solvent-accessible surface with key amino acid residues of Keap1, thus potentially regulating the Keap1-Nrf2 pathway by occupying the Nrf2-binding site on Keap1. Additionally, they exhibited strong cellular antioxidant activity and could protect HepG2 cells from AAPH-induced oxidative injury by reducing reactive oxygen species and MDA accumulation. This study firstly unraveled the molecular mechanisms of antioxidant peptides from FBBP, and provided a new theoretical basis for the high-value utilization of FBBP.

Nanoencapsulation of hybrid crude palm oil Unaué HIE OxG with jackfruit by-products as encapsulants: A study of cellular antioxidant activity and cytotoxicity in Caco-2 cells

Hybrid crude palm oil (HCPO) HIE OxG is notable for its abundance of carotenoids, tocopherols, and tocotrienols. Investigating cellular antioxidant activity (CAA) and the non-cytotoxicity of oil nanoparticles is crucial for understanding the behavior of these phytochemicals in biological systems and ensuring the safety of products. Nanoparticles of HCPO, encapsulated with jackfruit by-products were produced and characterized for CAA and cytotoxicity in Caco-2 cells. The nanoparticles exhibited nanoscale diameters (<250 nm), uniform distribution and stability (polydispersity index < 0.25; zeta potential JSF-NP −12.46 ± 0.15 mV and JAF-NP −13.73 ± 1.28 mV). JSF-NP and JAF-NP demonstrated superior CAA compared to the free HCPO across all concentrations, without inducing cytotoxic effects on differentiated Caco-2 cells. This study underscores the importance of investigating the CAA of edible oil nanoparticles, with non-cytotoxicity indicating biological safety and the potential to safeguard intestinal epithelial cells. Thus, JSF-NP and JAF-NP emerge as promising delivery systems for future HCPO applications.

Lipid composition micronutrient and cellular antioxidant properties of borage seed oil from eight provinces in China

AbstractBorage seed oil (BSO; Borago officinalis L.) has great medicinal and nutritional potential owing to its bioactive components. In this study, we compared the lipid composition, micronutrient content, free radical scavenging capacities, and cellular antioxidant and antiproliferative activities of BSO from eight provinces in China. The Gansu sample had the highest levels of γ‐linolenic acid, accounting for 18.66%. We detected more than 17 triacylglycerols in BSO. Among micronutrients, δ‐tocopherol and Δ5‐avenasterol were the main components of tocopherol and phytosterol, respectively. The highest cellular antioxidant activity values were found in the Shanxi and Gansu samples; the concentration cytotoxicity 50% values were higher than 200 mg/mL, and the median effective dose ranged from 18.34 ± 0.52 to 70.65 ± 1.76 mg/mL. In addition, we tested the free radical scavenging capacity of BSO and found significant differences in antioxidant capacity between samples from different regions, and the Gansu sample had a better antioxidant capacity.

Studying the Optimization, Characterization, and Antioxidant Activities of Phenolic Extracts Extracted from Rhus chinensis Mill. Leaf using Microwave-assisted Extraction System with Glycerol as a Green Solvent

Background: The leaves of Rhus chinensis Mill., a common deciduous tree found in the mild temperate zone of Asia, have many medicinal effects, including antioxidant properties. Objectives and Methods: This study aims to optimize the conditions for extracting phenols from Rhus chinensis Mill. (RCM) using a microwave-assisted extraction system with glycerol (MAEG) via response surface methodology (RSM). It also aims to compare the extraction efficacy of decoction and MAEG methods in terms of the bioactive compounds and antioxidant activities of the extracts obtained through them, identify bioactive compounds in both extracts via ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOFMS/ MS), and determine the cytotoxicity and cellular antioxidant activity of MAEG extract. Results: Temperature and glycerol concentration significantly affected the total phenolic content (TPC) of the extracts. The validated value of TPC was 84.11 ± 4.28 mg GAE/g for the sample obtained under the optimal conditions of 12.76 min at 54.08°C and 34.48% glycerol concentration. MAEG extract exhibited higher antioxidant properties compared to the decoction extract. Different phenolic compounds in the extracts were tentatively identified by LC-QTOF. MAEG concentrations from 1 mg/mL to 7.5 mg/mL were considered non-cytotoxic to NIH/3T3 fibroblasts. Furthermore, the cell viability of NIH/3T3 fibroblasts increased after being treated with MAEG extract (from 2.5 mg/mL to 7.5 mg/mL) and subjected to H2O2- induced oxidative stress compared to H2O2 treatment alone. Conclusion: Finally, MAEG can be used as a novel green extraction method for obtaining bioactive compounds for cosmetic and medicinal applications.

Exploring the novel antioxidant peptides in low-salt dry-cured ham: Preparation, purification, identification and molecular docking

Dry-cured ham is important source of bioactive peptides. In this study, the antioxidant activities of peptides and components from low and fully salted dry-cured hams were compared by peptidomics. And novel antioxidant peptides were identified and characterized. The results showed that the peptides (<3 KDa) extracted from low-salt dry-cured ham had higher antioxidant activity. Therefore, the antioxidant peptides in low-salt dry-cured ham were further characterized and the mechanism of their antioxidant activity was investigated. From the five candidate peptides selected, we found DWPDARGIWHND (DD12) to be highly stable, non-sensitizing, and non-toxic with the highest free radical scavenging activity. Molecular docking predicted that DD12 interacted with Keap1 through hydrogen-bond formation and hydrophobic interactions, suggesting that DD12 had good cellular antioxidant activity. DD12 peptide can bind to DPPH• and ABTS•+, resulting in strong free radical scavenging activity. Our findings support the development and application of natural antioxidant peptides in dry-cured ham.

A Culinary and Medicinal Gem: Exploring the Phytochemical and Functional Properties of Thai Basil.

This study aimed to comprehensively evaluate the potential health benefits of Thai basil extracts from two species, Ocimum basilicum var. thyrsiflorum and Ocimum basilicum cv. Jumbo 4320, by investigating their bioactivities, phytochemical composition, and in vitro antioxidant, antimicrobial, and antithrombotic activities. Thai basil extracts from two species (Ocimum basilicum var. thyrsiflorum and cv. Jumbo 4320) were obtained using water, ethanol, and ethyl acetate. Phytochemical analysis revealed chlorophylls, carotenoids, and diverse phenolic compounds. Its water extract boasted the highest total phenolics (459.62 ± 3.07 mg GAE/100 g), outperforming ethanol and ethyl acetate extracts (171.20 ± 1.10 and 66.02 ± 0.99 mg GAE/100 g, respectively). The ethanol extract of Jumbo 4320 also reigned supreme in total flavonoids (557.12 ± 14.27 mg CE/100 g), surpassing its counterparts (209.07 ± 23.31 and 131.41 ± 0.37 mg CE/100 g). O. basilicum cv. Jumbo 4320 extracts exhibited superior antioxidant activity by various assays. Jumbo 4320's water extract further dominated in the DPPH assay (IC50 48.52 ± 1.15 mg/mL), demonstrating superior free radical scavenging compared to ethanol and ethyl acetate extracts (IC50 60.54 ± 0.52 and 82.09 ± 3.18 mg/mL), respectively. While thyrsiflorum's ethanol extract claimed the top spot in the FRAP assay (0.0186 ± 0.00 mmol Fe2+/g), Jumbo 4320's water extract showcased the highest cellular antioxidant activity (80.62 ± 0.00% relative fluorescence intensity) in the DCFH-DA assay. Interestingly, Ocimum basilicum var. thyrsiflorum extracts demonstrated stronger antithrombotic activity at prolonging the prothrombin time at 78.3 ± 17.56 s. While the antimicrobial activity against most tested pathogens was limited, both species' extracts inhibited Bacillus cereus. These findings suggest the potential of Thai basil extracts, particularly from Jumbo 4320, as functional food ingredients with antioxidant and antithrombotic properties.

Glycation of tilapia protein hydrolysate decreases cellular antioxidant activity upon in vitro gastrointestinal digestion

Changes in structural characteristics and antioxidant activity of tilapia hydrolysate glycated with glucose, fructose, or xylose at 90 °C for 12 h, and following in vitro gastrointestinal (GI) digestion were investigated. Fourier-transformed infrared (FTIR) band between 1,800 and 1,400 cm−1 confirmed the structural modifications of hydrolysate under glycations. Glycation drastically increased ATBS·+ and ONOO- scavenging activities (p < 0.05) as well as ferric-reducing antioxidant power (FRAP). Xylose was the most effective sugar for glycation, yielding the highest chemical antioxidant activities (p < 0.05). However, glycated hydrolysates exhibited lower cellular antioxidant activity (CAA) on HepG2 cell when compared to hydrolysates. The extensive glycation of hydrolysates resulted in lower GI digestibility as confirmed by the FTIR spectra of CO, CN, NH, CC, CO, and CH stretching vibrations. Glycation of tilapia hydrolysates only improved chemical antioxidant activities, but alleviated CAA, especially upon simulated GI digestion.

Combination of Fenugreek and Quinoa Husk as Sources of Steroidal and Triterpenoid Saponins: Bioactivity of Their Co-Extracts and Hydrolysates.

Saponins, both steroidal and triterpenoid, exhibit distinct bioactivities. However, they are not commonly found together in natural sources; instead, sources tend to be rich in one type or another and mainly in the form of saponins rather than the sapogenin aglycones. Developing co-extracts containing both saponin or sapogenin types would be a strategy to harness their respective bioactivities, yielding multibioactive extracts. Therefore, this study evaluates the bioactivity (hypolipidemic, antioxidant, and anti-inflammatory activities) of co-extracts from fenugreek seeds (steroidal-rich saponins) and quinoa husk (triterpenoid-rich saponins), co-extracted at varying proportions, alongside their respective sapogenin-rich hydrolysates. Pancreatic lipase inhibition increased with fenugreek content in co-extracts, especially in sapogenin-rich variants. The latter substantially interfered with cholesterol bioaccessibility (90% vs. 15% in sapogenin-rich extracts). Saponin-rich co-extracts exhibited reduced cytokine release with increased fenugreek content, while sapogenin-rich counterparts showed greater reductions with higher quinoa husk content. Limited cellular antioxidant activities were observed in all extracts, with improved post-hydrolysis bioactivity. Therefore, simultaneous co-extraction of steroidal and triterpenoid sources, such as fenugreek and quinoa husk, as well as their subsequent hydrolysis, are innovative strategies for obtaining multibioactive natural extracts.

Plant-Derived Extracellular Vesicles: A New Revolutionization of Modern Healthy Diets and Biomedical Applications.

Plant-derived extracellular vesicles (PDEVs) have recently emerged as a promising area of research due to their potential health benefits and biomedical applications. Produced by various plant species, these EVs contain diverse bioactive molecules, including proteins, lipids, and nucleic acids. Increasing in vitro and in vivo studies have shown that PDEVs have inherent pharmacological activities that affect cellular processes, exerting anti-inflammatory, antioxidant, and anticancer activities, which can potentially contribute to disease therapy and improve human health. Additionally, PDEVs have shown potential as efficient and biocompatible drug delivery vehicles in treating various diseases. However, while PDEVs serve as a potential rising star in modern healthy diets and biomedical applications, further research is needed to address their underlying knowledge gaps, especially the lack of standardized protocols for their isolation, identification, and large-scale production. Furthermore, the safety and efficacy of PDEVs in clinical applications must be thoroughly evaluated. In this review, we concisely discuss current knowledge in the PDEV field, including their characteristics, biomedical applications, and isolation methods, to provide an overview of the current state of PDEV research. Finally, we discuss the challenges regarding the current and prospective issues for PDEVs. This review is expected to provide new insights into healthy diets and biomedical applications of vegetables and fruits, inspiring new advances in natural food-based science and technology.

Recent insights into catechins-rich Assam tea extract for photoaging and senescent ageing

Tea (Camellia spp.) is an important medicinal herb. C. sinensis var. sinensis is the most studied tea variety due to its more preferred flavor than C. sinensis var. assamica (Assam tea), the less economic importance with more bitter variety. A bitter taste highlights its potential as a candidate source for tea catechins, the health beneficial actives applicable for ageing treatment. Nonetheless, indicative data for tea on UV-induced and senescent ageing remain unclarified. Assam tea extract (ATE) was prepared and standardized in terms of TPC, TFC and TTC. EGCG was HPLC quantified as the prime ATE catechin. In vitro antioxidant activity of ATE was exhibited with ABTS, DPPH and FRAP assays. ATE’s cellular antioxidant activity was indicated in HDFs at a stronger degree than ascorbic acid. The photoaging protection of ATE was evidenced in a coculture of HaCaT cells and HDFs. ATE markedly suppressed UV-induced IL-6, IL-8, MMP-1 and MMP-9 expressions. The proficiency of ATE targeting on senescent ageing was demonstrated in an ex vivo human skin model, where IL-6 and MMP-1 expressions were suppressed, whilst hyaluronic acid and collagen syntheses were promoted. ATE was chemically stabled as indicated by the catechin contents and color parameters following 6 months storage under conditions recommended for topical product. ATE enriched in catechins warrants its applicability as a new generation of photoaging protectant agent promising for the prevention and treatment for senescent ageing. The findings indicate the proficiency of ATE for innovative anti-ageing agent.

Olive (Olea europaea L.) Seed as New Source of Cholesterol-Lowering Bioactive Peptides: Elucidation of Their Mechanism of Action in HepG2 Cells and Their Trans-Epithelial Transport in Differentiated Caco-2 Cells.

The production of olive oil has important economic repercussions in Mediterranean countries but also a considerable impact on the environment. This production generates enormous quantities of waste and by-products, which can be exploited as new raw materials to obtain innovative ingredients and therefore make the olive production more sustainable. In a previous study, we decided to foster olive seeds by generating two protein hydrolysates using food-grade enzymes, alcalase (AH) and papain (PH). These hydrolysates have shown, both in vitro and at the cellular level, antioxidant and antidiabetic activities, being able to inhibit the activity of the DPP-IV enzyme and modulate the secretion of GLP-1. Given the multifunctional behavior of peptides, both hydrolysates displayed dual hypocholesterolemic activity, inhibiting the activity of HMGCoAR and impairing the PPI of PCSK9/LDLR, with an IC50 equal to 0.61 mg/mL and 0.31 mg/mL for AH and PH, respectively. Furthermore, both samples restored LDLR protein levels on the membrane of human hepatic HepG2 cells, increasing the uptake of LDL from the extracellular environment. Since intestinal bioavailability is a key component of bioactive peptides, the second objective of this work is to evaluate the capacity of AH and PH peptides to be transported by differentiated human intestinal Caco-2 cells. The peptides transported by intestinal cells have been analyzed using mass spectrometry analysis, identifying a mixture of stable peptides that may represent new ingredients with multifunctional qualities for the development of nutraceuticals and functional foods to delay the onset of metabolic syndrome, promoting the principles of environmental sustainability.

The influence of unique interfacial networks based on egg white proteins for the stabilization of high internal phase Pickering emulsions: Physical stability and free fatty acid release kinetics

This study was oriented towards the impacts of unique interfacial networks, formed by glycosylated and non-glycosylated egg white proteins, on the characteristics of high internal phase Pickering emulsions (HIPPEs). Glycosylated egg white protein particles (EWPG) manifested a more compact protein tertiary structure and amplified surface hydrophobicity, forming durable coral-like networks at the oil-water interface. The non-glycosylated egg white protein particles (EWP) could form spherical cluster interfacial networks. Raman spectroscopy analysis illuminated that EWPG could exhibit better interactions with aliphatic amino acids via hydrogen bonds and hydrophobic interactions. The release of free fatty acid (FFA) from both HIPPEs followed the first-order kinetic model with a combination of diffusion. EWPG-stabilized HIPPEs demonstrated superior physical stability and cellular antioxidant activity. This research shed light on the promising prospects of HIPPEs as promising amphiphilic delivery systems with capabilities to co-deliver hydrophilic and hydrophobic nutraceuticals and amplify their intracellular biological potency.

Antibiofilm, cellular antioxidant, anti-inflammatory, immunomodulatory, cytotoxic, and antimutagenic activities of soluble melanins from Randia echinocarpa fruit

Antibiofilm, cellular antioxidant, anti-inflammatory, immunomodulatory, cytotoxic, and antimutagenic activities of soluble melanins from Randia echinocarpa fruit

Anti-Cancer and Anti-Oxidant Bioactive Metabolites from Aspergillus fumigatus WA7S6 Isolated from Marine Sources: In Vitro and In Silico Studies.

Cancer is a huge global disease burden. Every year, tens of millions of people worldwide are diagnosed with cancer, and more than half of them die as a result of it. The great biodiversity of the marine environment has increasingly piqued the interest of experts, especially in the field of drug discovery. The marine fungus Aspergillus fumigatus WA7S6 has been selected among a group of fungi isolated from marine sponges as it exhibits a pronounced antimicrobial activity toward a group of pathogenic microbes. The fungus has been identified genetically by amplification and analysis of its 18srRNA gene. The fungus crude extract has been obtained by cultivation of the fungus on rice media. The crude extract was tested for antibacterial activity against a variety of pathogenic microorganisms. The results demonstrated a pronounced antimicrobial action against P. aeruginosa, S. aureus, A. niger, and Candida albicans. Furthermore, we tested the antioxidant potential of the Aspergillus fumigatus WA7S6 crude extract using three different methods: ATBS, DPPH, and lipid peroxidation assays. Results showed that the crude extract WA7S6 had an IC50 value of 21.35 µg/mL. The anticancer potential of the crude extract was also evaluated against cancer cell lines such as Hela, MCF, and WI-38. The chemical profiling of the fungus extract was identified via GC-mass and in silico molecular docking of the identified compounds on heme oxygenase, as a stress protein included in cellular protection, antioxidant, and anti-inflammatory activities, suggesting that some compounds, such as 9-Tetradecynoic acid, 11-Hexadecynoic acid, methyl ester, and dehydromevalonic lactone, could be relevant for antioxidant purposes.

Anthocyanin-rich extract from purple tea: Chemical stability, cellular antioxidant activity, and protection of human plasma

This study aimed to obtain an anthocyanin extract from the purple leaves of Camellia sinensis cv. Zijuan using a sustainable, non-toxic, and low-cost solid-liquid extraction, employing an aqueous citric acid solution (0.2 mol/L) as the extracting solvent, and to evaluate its chemical stability at different pH values, as well as its in vitro antioxidant properties in chemical and biological terms. The phenolic composition, in vitro antioxidant activity, and the stability of anthocyanins against pH, temperature, and light of the crude extract (CE) were evaluated, as well as the phenolic composition and bioactivity in the crude lyophilised extract (CLE). In the direct/reverse spectrophotometric titration, anthocyanins showed structural changes between pH 2 and 10, and reversibility of 80%. The antioxidant activity against the DPPH radical showed inhibition percentages of 73% (pH 4.5) to 39% (pH 10). Thermal stability was observed at 60 °C, and prolonged exposure of the extract to light caused photodegradation of the anthocyanins. Thirty-three phenolic compounds, including anthocyanins and catechins, were quantified in the CLE by UPLC-ESI-MS and HPLC, totalling 40.18 mg/g. CLE reduced cell viability (IC50 from 18.1 to 52.5 μg GAE/mL), exerted antiproliferative (GI50 from 0.0006 to 17.0 μg GAE/mL) and cytotoxic (LC50 from 33.2 to 89.9 μg GAE/mL) effects against A549 (human lung adenocarcinoma epithelial cells), HepG2 (hepatocellular carcinoma), HCT8 (ileocecal colorectal adenocarcinoma), and Eahy926 (somatic cell hybrid cells); and showed protection against oxidation of human plasma (635 ± 30 mg AAE/g). The results showed the diversity of compounds in the extracts and their potential for technological applications; however, temperature, pH, and light must be considered to avoid diminishing their bioactivity.

Sulfated polysaccharides from Caulerpa lentillifera: Optimizing the process of extraction, structural characteristics, antioxidant capabilities, and anti-glycation properties

The polysaccharides found in Caulerpa lentillifera (sea grape algae) are potentially an important bioactive resource. This study makes use of RSM (response surface methodology) to determine the optimal conditions for the extraction of valuable SGP (sea grape polysaccharides). The findings indicated that a water/raw material ratio of 10:1 mL/g, temperature of 90 °C, and extraction time of 45 min would maximize the yield, with experimentation achieving a yield of 21.576 %. After undergoing purification through DEAE-52 cellulose and Sephacryl S-100 column chromatography, three distinct fractions were obtained, namely SGP11, SGP21, and SGP31, each possessing average molecular weights of 38.24 kDa, 30.13 kDa, and 30.65 kDa, respectively. Following characterization, the fractions were shown to comprise glucose, galacturonic acid, xylose, and mannose, while the sulfate content was in the range of 12.2–21.8 %. Using Fourier transform infrared spectroscopy (FT-IR) it was possible to confirm with absolute certainty the sulfate polysaccharide attributes of SGP11, SGP21, and SGP31. NMR (nuclear magnetic resonance) findings made it clear that SGP11 exhibited α-glycosidic configurations, while the configurations of SGP21 and SGP31 were instead β-glycosidic. The in vitro antioxidant assays which were conducted revealed that each of the fractions was able to demonstrate detectable scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cations. All fractions were also found to exhibit the capacity to scavenge NO radicals in a dose-dependent manner. SGP11, SGP21, and SGP31 were also able to display cellular antioxidant activity (CAA) against the human adenocarcinoma colon (Caco-2) cell line when oxidative damage was induced. The concentration levels were found to govern the extent of such activity. Moreover, purified SGP were found to exert strong inhibitory effects upon glycation, with the responses dependent upon dosage, thus confirming the potential for SGP to find a role as a natural resource for the production of polysaccharide-based antioxidant drugs, or products to promote improved health.

The antioxidant peptides from walnut protein hydrolysates and their protective activity against alcoholic injury.

In this study, walnut protein was hydrolyzed, separated by ultrafiltration, purified by RP-HPLC, identified by LC-MS/MS, and screened by molecular docking to finally obtain three novel antioxidant peptides HGEPGQQQR (1189.584 Da), VAPFPEVFGK (1089.586 Da) and HNVADPQR (949.473 Da). These three peptides exhibited excellent cellular antioxidant activity (CAA) with EC50 values of 0.0120 mg mL-1, 0.0068 mg mL-1, and 0.0069 mg mL-1, respectively, which were superior to that of the positive control GSH (EC50: 0.0122 mg mL-1). In the ethanol injury model, three antioxidant peptides enhanced the survival of cells treated with ethanol from 47.36% to 62.69%, 57.06% and 71.64%, respectively. Molecular docking results showed that the three antioxidant peptides could effectively bind to Keap1, CYP2E1 and TLR4 proteins. These results suggested that walnut-derived antioxidant peptides could be potential antioxidants and hepatoprotective agents for application in functional foods.

Nutritional profiling, fiber content and in vitro bioactivities of wheat-based biscuits formulated with novel ingredients.

This study evaluated the nutritional profile and fiber content of innovative formulations of wheat-based biscuits enriched with chia seeds, carob flour and coconut sugar. The in vitro antioxidant, cytotoxic, anti-inflammatory and antimicrobial activities were also investigated to understand the potential health advantages of the incorporation of these new ingredients. The novel biscuits demonstrated significant improvements in protein and mineral content, with increases of 50% and 100% in chia biscuits, and up to 20% and 40% in carob biscuits, respectively. Fiber also notably increased, particularly in samples containing 10% carob flour, which increased four times as compared to wheat-based samples. The new ingredients exhibited antibacterial and antifungal activity, particularly against Yersinia enterocolitica (minimum inhibitory concentration 1.25 mg mL-1 in coconut sugar) and Aspergillus fumigatus (minimum inhibitory concentration/minimum fungicidal concentrations 2.5/5 mg mL-1 in chia seeds). However, the final biscuits only displayed antifungal properties. Carob flour and chia seeds had a remarkably high capacity to inhibit the formation of TBARS and promoted greater antioxidant activity in biscuit formulations, with EC50 values decreasing from 23.25 mg mL-1 (control) to 4.54 mg mL-1 (15% defatted ground chia seeds) and 1.19 mg mL-1 (10% carob flour). Only chia seeds exhibited cellular antioxidant, anti-inflammatory and cytotoxic activity, attributes that were lost when seeds were added into the biscuits. These findings highlight the potential health benefits of these ingredients, particularly when incorporated in new wheat-based formulations.

Antioxidant interaction between α-tocopherol and γ-oryzanol in HepG2 cells.

Minor constituents exhibit certain antioxidant interactions in vitro, and the effects in different media are different. However, it is not clear whether there are antioxidant interactions in cells after digestion and absorption. We utilized the cellular antioxidant evaluation model in HepG2 cells to study the antioxidant interaction between α-tocopherol and γ-oryzanol, and the interaction mechanism of a binary mixture was also illustrated. A cellular antioxidant assay (CAA) model and a combined index (CI) method were firstly used to explore the antioxidant activity and interaction of the binary mixture in HepG2 cells. The CAA value was positively correlated with the single addition concentration, while the results displayed a biphasic tendency with increasing concentrations of the binary mixture. The combination of TO11 (1 μg mL-1 α-tocopherol and 10 μg mL-1 γ-oryzanol) showed the greatest antioxidant activity and synergistic effect, and the maximum CAA value reached up to 94.84 ± 4.2. Then the mechanism of the synergistic antioxidant effect of the binary mixture was explained from three aspects including cellular uptake, intracellular reactive oxygen species (ROS) level and endogenous enzyme activity. The results demonstrated that the antioxidant interaction of the binary mixture in cells was related to cellular uptake of minor constituents, and the combination of TO11 exerted a synergistic effect by scavenging ROS and up-regulating glutathione peroxidase (GSH-Px) activity, resulting in the strongest cellular antioxidant activity. This study throws light on the nature of antioxidant interaction between minor constituents, which may contribute to the development of related functional foods and rational dietary collocation.