Antioxidants

15 pages, 893 KiB

Open AccessArticle

Postharvest Treatment with Abscisic Acid Alleviates Chilling Injury in Zucchini Fruit by Regulating Phenolic Metabolism and Non-Enzymatic Antioxidant System

by Alejandro Castro-Cegrí, Sandra Sierra, Laura Hidalgo-Santiago, Adelaida Esteban-Muñoz, Manuel Jamilena, Dolores Garrido and Francisco Palma

Antioxidants 2023, 12(1), 211; https://doi.org/10.3390/antiox12010211 - 16 Jan 2023

Cited by 20 | Viewed by 4217

Abstract

Reports show that phytohormone abscisic acid (ABA) is involved in reducing zucchini postharvest chilling injury. During the storage of harvested fruit at low temperatures, chilling injury symptoms were associated with cell damage through the production of reactive oxygen species. In this work, we [...] Read more.

Reports show that phytohormone abscisic acid (ABA) is involved in reducing zucchini postharvest chilling injury. During the storage of harvested fruit at low temperatures, chilling injury symptoms were associated with cell damage through the production of reactive oxygen species. In this work, we have studied the importance of different non-enzymatic antioxidants on tolerance to cold stress in zucchini fruit treated with ABA. The application of ABA increases the antioxidant capacity of zucchini fruit during storage through the accumulation of ascorbate, carotenoids and polyphenolic compounds. The quantification of specific phenols was performed by UPLC/MS-MS, observing that exogenous ABA mainly activated the production of flavonoids. The rise in all these non-enzymatic antioxidants due to ABA correlates with a reduction in oxidative stress in treated fruit during cold stress. The results showed that the ABA mainly induces antioxidant metabolism during the first day of exposure to low temperatures, and this response is key to avoiding the occurrence of chilling injury. This work suggests an important protective role of non-enzymatic antioxidants and polyphenolic metabolism in the prevention of chilling injury in zucchini fruit. Full article

(This article belongs to the Special Issue Biological Potential of Antioxidant Compounds from Vegetable Sources)

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20 pages, 1396 KiB

Open AccessArticle

Post-Distillation By-Products of Aromatic Plants from Lamiaceae Family as Rich Sources of Antioxidants and Enzyme Inhibitors

by Simon Vlad Luca, Gokhan Zengin, Kouadio Ibrahime Sinan, Krystyna Skalicka-Woźniak and Adriana Trifan

Antioxidants 2023, 12(1), 210; https://doi.org/10.3390/antiox12010210 - 16 Jan 2023

Cited by 8 | Viewed by 2933

Abstract

There is currently no use for the vast quantities of post-distillation by-products, such as spent plant materials and residual waters, produced by the essential oil (EO) industry of aromatic herbs. In this study, the EOs of three Lamiaceae species (thyme, oregano, and basil) [...] Read more.

There is currently no use for the vast quantities of post-distillation by-products, such as spent plant materials and residual waters, produced by the essential oil (EO) industry of aromatic herbs. In this study, the EOs of three Lamiaceae species (thyme, oregano, and basil) and their total, spent, and residual water extracts were phytochemically characterized and biologically assessed. The collected information was put through a series of analyses, including principal component analysis, heatmap analysis, and Pearson correlation analysis. Concerning the EOs, 58 volatile compounds were present in thyme (e.g., p-cymene, thymol), 44 compounds in oregano (e.g., thymol, carvacrol), and 67 compounds in basil (e.g., eucalyptol, linalool, estragole, (E)-methyl cinnamate). The LC-HRMS/MS analysis of the total, spent, and residual water extracts showed the presence of 31 compounds in thyme (e.g., quercetin-O-hexoside, pebrellin, eriodictyol), 31 compounds in oregano (e.g., rosmarinic acid, apigenin, kaempferol, salvianolic acids I, B, and E), and 25 compounds in basil (e.g., fertaric acid, cichoric acid, caftaric acid, salvianolic acid A). The EOs of the three Lamiaceae species showed the highest metal-reducing properties (up to 1792.32 mg TE/g in the CUPRAC assay), whereas the spent extracts of oregano and basil displayed very high radical-scavenging properties (up to 266.59 mg TE/g in DPPH assay). All extracts exhibited anti-acetylcholinesterase (up to 3.29 mg GALAE/g), anti-tyrosinase (up to 70.00 mg KAE/g), anti-amylase (up to 0.66 mmol ACAE/g), and anti-glucosidase (up to 1.22 mmol ACAE/g) effects. Thus, the present research demonstrated that both the raw extracts (EOs and total extracts) and the post-distillation by-products (spent material and residual water extracts) are rich in bioactive metabolites with antioxidant and enzyme inhibitory properties. Full article

(This article belongs to the Special Issue Antioxidant and Biological Properties of Plant Extracts II)

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19 pages, 2477 KiB

Open AccessArticle

Dietary Methionine Level Impacts the Growth, Nutrient Metabolism, Antioxidant Capacity and Immunity of the Chinese Mitten Crab (Eriocheir sinensis) under Chronic Heat Stress

by Jiadai Liu, Cong Zhang, Xiaodan Wang, Xinyu Li, Qincheng Huang, Han Wang, Yixin Miao, Erchao Li, Jianguang Qin and Liqiao Chen

Antioxidants 2023, 12(1), 209; https://doi.org/10.3390/antiox12010209 - 16 Jan 2023

Cited by 10 | Viewed by 2494

Abstract

This study examined whether diets with high dietary methionine levels could alleviate chronic heat stress in Chinese mitten crab Eriocheir sinensis. Crabs were fed three dietary methionine levels of 0.49%, 1.29% and 2.09% for six weeks. The analyzed methionine concentration of diets [...] Read more.

This study examined whether diets with high dietary methionine levels could alleviate chronic heat stress in Chinese mitten crab Eriocheir sinensis. Crabs were fed three dietary methionine levels of 0.49%, 1.29% and 2.09% for six weeks. The analyzed methionine concentration of diets was 0.48%, 1.05% and 1.72%, respectively. Crabs were fed three different supplemental concentrations of dietary methionine at 24 °C and 30 °C, respectively. The trial was divided into six groups with five replicates in each group, and 40 juvenile crabs (initial average weight 0.71 ± 0.01 g) in each replicate. During the trial, crabs were fed twice daily (the diet of 4% of the body weight was delivered daily). The effects of dietary methionine level on nutrient metabolism, antioxidant capacity, apoptosis factors and immunity were evaluated at a normal water temperature of 24 °C and high temperature of 30 °C. Feed conversion ratio decreased under chronic heat stress. Chronic heat stress increased weight gain, specific growth rate, molting frequency, and protein efficiency ratio. The survival of crabs decreased under chronic heat stress, whereas a high level of dietary methionine significantly improved survival. Chronic heat stress induced lipid accumulation and protein content reduction. The high-methionine diet decreased lipid in the body and hepatopancreas, but increased protein in the body, muscle and hepatopancreas under chronic heat stress. Simultaneously, the high dietary methionine levels mitigated oxidative stress by reducing lipid peroxidation, restoring the antioxidant enzyme system, decreasing apoptosis and activating immune function under chronic heat stress. This study suggests that supplementing 1.72% dietary methionine could alleviate the adverse effects of a high water temperature in E. sinensis farming. Full article

(This article belongs to the Topic Antioxidant Activity of Natural Products)

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14 pages, 299 KiB

Open AccessArticle

Effects of Dietary Supplementation of Pomegranate Peel with Xylanase on Egg Quality and Antioxidant Parameters in Laying Hens

by Styliani Lioliopoulou, Georgios A. Papadopoulos, Ilias Giannenas, Konstantina Vasilopoulou, Clare Squires, Paschalis Fortomaris and Fani Th. Mantzouridou

Antioxidants 2023, 12(1), 208; https://doi.org/10.3390/antiox12010208 - 16 Jan 2023

Cited by 12 | Viewed by 2590

Abstract

Pomegranate contains bioactive compounds in all its parts. In this study, two levels of pomegranate peel byproduct (PPB) with or without the inclusion of xylanase enzyme were used to supplement laying hens’ diet, in a 2 × 2 full factorial design. A total [...] Read more.

Pomegranate contains bioactive compounds in all its parts. In this study, two levels of pomegranate peel byproduct (PPB) with or without the inclusion of xylanase enzyme were used to supplement laying hens’ diet, in a 2 × 2 full factorial design. A total of 48 Isa brown laying hens were fed the following experimental diets for 8 weeks: T1 (2.5% PPB); T2 (2.5% PPB and xylanase); T3 (5% PPB); T4 (5% PPB and xylanase). Eggs collected were analyzed for egg quality parameters. Moreover, egg yolks were analyzed for Malondialdehyde content (MDA), fatty acid profile and total phenolic content. The T2 eggs showed enhanced yolk coloration and greater yolk total phenolic content. The T3 and T4 egg yolks showed lower MDA levels compared with T1, T2. Overall, results have shown that (a) xylanase inclusion affected egg yolk coloration and total phenolic content when combined with 2.5% PPB dietary supplementation; (b) dietary supplementation of 5% PPB resulted in eggs with reduced MDA levels. Full article

(This article belongs to the Special Issue Oxidative Stress, Reactive Oxygen Species and Animal Nutrition)

12 pages, 2056 KiB

Open AccessArticle

Facile Fabrication of α-Bisabolol Nanoparticles with Improved Antioxidant and Antibacterial Effects

by Sangwoo Kim, Sohyeon Yu, Jisu Kim, Nisar Ul Khaliq, Won Il Choi, Hyungjun Kim and Daekyung Sung

Antioxidants 2023, 12(1), 207; https://doi.org/10.3390/antiox12010207 - 16 Jan 2023

Cited by 5 | Viewed by 3228

Abstract

Bioactive compounds are widely used in the bio-industry because of their antioxidant and antibacterial activities. Because of excessive oxidative stress, which causes various diseases in humans, and because preservatives used in bioproducts cause allergies and contact dermatitis, it is important to use natural [...] Read more.

Bioactive compounds are widely used in the bio-industry because of their antioxidant and antibacterial activities. Because of excessive oxidative stress, which causes various diseases in humans, and because preservatives used in bioproducts cause allergies and contact dermatitis, it is important to use natural bioactive compounds in bioproducts to minimize oxidative stress. α-bisabolol (ABS) is a natural compound with both antioxidant and antibacterial properties. However, its water-insolubility makes its utilization in bioproducts difficult. In this study, ABS-loaded polyglyceryl-4 caprate nanoparticles (ABS@NPs) with improved aqueous stability and ABS loading were fabricated using an encapsulation method. The long-term stability of the ABS@NPs was analyzed with dynamic light scattering and methylene blue-staining to determine the optimized ABS concentration in ABS@NPs (10 wt%). The ABS@NPs exhibited excellent antioxidant activity, according to the 2,2-diphenyl-1-picrylhydrazyl assay and in vitro reactive oxygen species generation in NIH-3T3 fibroblast cells, and an outstanding antibacterial effect, as determined using the Staphylococcus aureus colony-counting method. Furthermore, we evaluated the biocompatibility of the ABS@NPs in vitro. This study suggests that ABS@NPs with improved antioxidant and antibacterial properties can be used to treat diseases related to various oxidative stresses and can be applied in many fields, such as pharmaceuticals, cosmetics, and foods. Full article

(This article belongs to the Special Issue Applications of Antioxidant Nanoparticles)

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Schematic illustration of the preparation of α -bisabolol-loaded nanoparticles (ABS@NPs) and improved antioxidant and antibacterial effects. Full article ">Figure 2
(A) Particle size, (B) polydispersity index (PDI), and (C) zeta potential of ABS@NPs with different ABS loadings in the range of 0 to 40 wt%. (D) Methylene blue staining test of the ABS@NPs with different ABS loadings. Full article ">Figure 3
Long-term stability of ABS@NPs at ABS concentrations ranging from 0 to 40 wt%. Variation of the (A) particle size and (B) polydispersity index (PDI) of ABS@NPs over 112 days. Full article ">Figure 4
(A) Antioxidation activity of ABS@NPs with 0, 5, and 10 wt% ABS, compared to that of ascorbic acid (AA) in deionized water (DIW), assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay. (B) Results of in vitro assays using the H2DCFDA assay kit. Antioxidation activity of ABS@NP 10 wt% at ABS concentrations ranging from 1 to 1000 nM; reactive oxygen species (ROS) and the control (CTL) groups represent the highest and lowest levels of ROS, respectively (* p < 0.05, ** p < 0.01, *** p < 0.005). Full article ">Figure 5
Antibacterial activity of ABS@NPs. (A) Photographs of staphylococcus aureus ATCC 6538 colonies in (i) phosphate-buffered saline, (ii) ABS in deionized water (DIW), (iii) ABS@NP 0 wt% in DIW, and (iv) ABS@NP 10 wt% in DIW. (B) Viability of S. aureus after treatment with phosphate-buffered saline (PBS), ABS in DIW, ABS@NP 0 wt% in DIW, and ABS@NP 10 wt% in DIW. Full article ">Figure 6
Cytotoxicity analysis of 10 wt% ABS@NPs at ABS concentrations in the range of 1 to 10 μM (* p < 0.05). Full article ">

11 pages, 1752 KiB

Open AccessArticle

Effect of Ascosphaera apis Infestation on the Activities of Four Antioxidant Enzymes in Asian Honey Bee Larval Guts

by Kaiyao Zhang, Zhongmin Fu, Xiaoxue Fan, Zixin Wang, Siyi Wang, Sijia Guo, Xuze Gao, Haodong Zhao, Xin Jing, Peiyuan Zou, Qiming Li, Mengjun Chen, Dafu Chen and Rui Guo

Antioxidants 2023, 12(1), 206; https://doi.org/10.3390/antiox12010206 - 16 Jan 2023

Cited by 2 | Viewed by 2886

Abstract

Ascosphaera apis infects exclusively bee larvae and causes chalkbrood, a lethal fungal disease that results in a sharp reduction in adult bees and colony productivity. However, little is known about the effect of A. apis infestation on the activities of antioxidant enzymes in [...] Read more.

Ascosphaera apis infects exclusively bee larvae and causes chalkbrood, a lethal fungal disease that results in a sharp reduction in adult bees and colony productivity. However, little is known about the effect of A. apis infestation on the activities of antioxidant enzymes in bee larvae. Here, A. apis spores were purified and used to inoculate Asian honey bee (Apis cerana) larvae, followed by the detection of the host survival rate and an evaluation of the activities of four major antioxidant enzymes. At 6 days after inoculation (dpi) with A. apis spores, obvious symptoms of chalkbrood disease similar to what occurs in Apis mellifera larvae were observed. PCR identification verified the A. apis infection of A. cerana larvae. Additionally, the survival rate of larvae inoculated with A. apis was high at 1–2 dpi, which sharply decreased to 4.16% at 4 dpi and which reached 0% at 5 dpi, whereas that of uninoculated larvae was always high at 1~8 dpi, with an average survival rate of 95.37%, indicating the negative impact of A. apis infection on larval survival. As compared with those in the corresponding uninoculated groups, the superoxide dismutase (SOD) and catalase (CAT) activities in the 5- and 6-day-old larval guts in the A. apis–inoculated groups were significantly decreased (p < 0.05) and the glutathione S-transferase (GST) activity in the 4- and 5-day-old larval guts was significantly increased (p < 0.05), which suggests that the inhibition of SOD and CAT activities and the activation of GST activity in the larval guts was caused by A. apis infestation. In comparison with that in the corresponding uninoculated groups, the polyphenol oxidase (PPO) activity was significantly increased (p < 0.05) in the 5-day-old larval gut but significantly reduced (p < 0.01) in the 6-day-old larval gut, indicating that the PPO activity in the larval guts was first enhanced and then suppressed. Our findings not only unravel the response of A. cerana larvae to A. apis infestation from a biochemical perspective but also offer a valuable insight into the interaction between Asian honey bee larvae and A. apis. Full article

(This article belongs to the Section Antioxidant Enzyme Systems)

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22 pages, 2538 KiB

Open AccessArticle

Valorization of Peels of Eight Peach Varieties: GC–MS Profile, Free and Bound Phenolics and Corresponding Biological Activities

by Dasha Mihaylova, Aneta Popova, Ivelina Desseva, Ivayla Dincheva and Yulian Tumbarski

Antioxidants 2023, 12(1), 205; https://doi.org/10.3390/antiox12010205 - 16 Jan 2023

Cited by 9 | Viewed by 2547

Abstract

Sustainability, becoming essential for food processing and technology, sets goals for the characterization of resources considered as food waste. In this work, information about the GC-MS metabolites of peach peels was provided as a tool that can shed more light on the studied [...] Read more.

Sustainability, becoming essential for food processing and technology, sets goals for the characterization of resources considered as food waste. In this work, information about the GC-MS metabolites of peach peels was provided as a tool that can shed more light on the studied biological activities. In addition, distribution patterns and contribution of the chemical profile and free and bound phenolic compounds as antioxidant, antimicrobial, and enzymatic clusters in peach peels of different varieties of Bulgarian origin were studied. The two applied techniques (alkaline and acid hydrolysis) for releasing the bound phenolics reveal that alkaline hydrolysis is a better extraction approach. Still, the results indicate the prevalence of the free phenolics in the studied peach peel varieties. Total phenolics of peach wastes were positively correlated with their antioxidant activity. The antioxidant activity results certainly defined the need of an individual interpretation for each variety, but the free phenolics fractions could be outlined with the strongest potential. The limited ability of the peels’ extracts to inhibit α-amylase and acetylcholinesterase, and the moderate antimicrobial activity, on the other hand, indicate that the potential of peach peels is still sufficient to seek ways to valorize this waste. Indeed, this new information about peach peels can be used to characterize peach fruits from different countries and/or different food processes, as well as to promote the use of this fruit waste in food preparation. Full article

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Free and bound (insoluble) phenolics content distribution of eight peach varieties’ peel extracts—(A) Total phenolic content (TPC, mgGAE/g dw), (B) Total flavonoids (TFC, μgQE/g dw) and (C) Total monomeric anthocyanins (TMA, µg cyanidin-3-glucoside (C3GE)/g dw). G—“Gergana”, F—“Filina”, U—“Ufo 4”, JL—“July Lady”, L—“Laskava”, FQ—“Flat queen”, Evm—“Evmolpiya”, M—“Morsiani 90”. Different letters (a–l) within chart columns indicate significant differences (p < 0.05) between treatments as analyzed by two-way ANOVA and the Tukey test (n = 3 per treatment group). Full article ">Figure 1 Cont.
Free and bound (insoluble) phenolics content distribution of eight peach varieties’ peel extracts—(A) Total phenolic content (TPC, mgGAE/g dw), (B) Total flavonoids (TFC, μgQE/g dw) and (C) Total monomeric anthocyanins (TMA, µg cyanidin-3-glucoside (C3GE)/g dw). G—“Gergana”, F—“Filina”, U—“Ufo 4”, JL—“July Lady”, L—“Laskava”, FQ—“Flat queen”, Evm—“Evmolpiya”, M—“Morsiani 90”. Different letters (a–l) within chart columns indicate significant differences (p < 0.05) between treatments as analyzed by two-way ANOVA and the Tukey test (n = 3 per treatment group). Full article ">Figure 2
Antioxidant activity of free and bound phenolics in eight peach varieties’ peel extracts (µMTE/g dw) by (A) DPPH, (B) ABTS, (C) FRAP and (D) CUPRAC assays. Different letters (a–n) within chart columns indicate significant differences (p < 0.05) between treatments as analyzed by two-way ANOVA and the Tukey test (n = 3). G—“Gergana”, F—“Filina”, U—“Ufo 4”, JL—“July Lady”, L—“Laskava”, FQ—“Flat queen”, Evm—“Evmolpiya”, M—“Morsiani 90”. Full article ">Figure 2 Cont.
Antioxidant activity of free and bound phenolics in eight peach varieties’ peel extracts (µMTE/g dw) by (A) DPPH, (B) ABTS, (C) FRAP and (D) CUPRAC assays. Different letters (a–n) within chart columns indicate significant differences (p < 0.05) between treatments as analyzed by two-way ANOVA and the Tukey test (n = 3). G—“Gergana”, F—“Filina”, U—“Ufo 4”, JL—“July Lady”, L—“Laskava”, FQ—“Flat queen”, Evm—“Evmolpiya”, M—“Morsiani 90”. Full article ">Figure 3
Principal component score plot (A) and eigenvector load values (B) of GC-MS data of volatile compounds of peach (Prunus persica L.) peels for the eight peach peel varieties. Full article ">Figure 4
Principal component score plot (A) and eigenvector load values (B) of TPC, TMA, TFC, and AOA assays of peach (Prunus persica L.) peels for the eight peach peel varieties. Full article ">Figure 5
Heatmap of the clustering result of peach peels from eight varieties. (A) GC-MS data of volatile compounds and (B) TPC, TMA, TFC, and AOA assays. Values were normalized by log10 transformation. Full article ">

15 pages, 3625 KiB

Open AccessArticle

Antioxidant and Anti-Inflammatory Effects of 6,3’,4´- and 7,3´,4´-Trihydroxyflavone on 2D and 3D RAW264.7 Models

by Xiang Wang, Yujia Cao, Siyu Chen, Xin Yang, Jinsong Bian and Dejian Huang

Antioxidants 2023, 12(1), 204; https://doi.org/10.3390/antiox12010204 - 16 Jan 2023

Cited by 5 | Viewed by 2571

Abstract

Dietary flavones 6,3´,4´-trihydroxyflavone (6,3´,4´-HOFL) and 7,3´,4´-trihydroxyflavone (7,3´,4´-HOFL) showed preliminary antioxidant and anti-inflammatory activities in a two-dimensional (2D) cell culture model. However, their action mechanisms remain unclear, and the anti-inflammatory activities have not been studied in a reliable three-dimensional (3D) cell model. Therefore, in [...] Read more.

Dietary flavones 6,3´,4´-trihydroxyflavone (6,3´,4´-HOFL) and 7,3´,4´-trihydroxyflavone (7,3´,4´-HOFL) showed preliminary antioxidant and anti-inflammatory activities in a two-dimensional (2D) cell culture model. However, their action mechanisms remain unclear, and the anti-inflammatory activities have not been studied in a reliable three-dimensional (3D) cell model. Therefore, in the current study, the antioxidant potency was examined by their scavenging ability of cellular reactive oxygen species. Anti-inflammatory activities were examined via their inhibitory effects on inflammatory mediators in vitro on 2D and 3D macrophage models, and their mechanisms were determined through transcriptome. In the 3D macrophages, two flavones were less bioactive than they were in 2D macrophages, but they both significantly suppressed the overexpression of proinflammatory mediators in two cell models. The divergent position of the hydroxyl group on the A ring resulted in activity differences. Compared to 6,3´,4´-HOFL, 7,3´,4´-HOFL showed lower activity on NO, IL-1β suppression, and c-Src binding (IC₅₀: 12.0 and 20.9 µM) but higher ROS-scavenging capacity (IC₅₀: 3.20 and 2.71 µM) and less cytotoxicity. In addition to the IL-17 and TNF pathways of 6,3´,4´-HOFL, 7,3´,4´-HOFL also exerted anti-inflammatory activity through JAK-STAT, as indicated by the RNA-sequencing results. Two flavones showed prominent antioxidant and anti-inflammatory activities on 2D and 3D models. Full article

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ROS-scavenging activity of 6,3´,4´- and 7,3´,4´-HOFL in tBHP-induced RAW264.7. Fluorescent microscopy images of untreated (a), tBHP-induced (b), and tBHP- and 6,3´,4´-HOFL- (c) or tBHP and 7,3´,4´-HOFL-treated cells (d). Blue: cell nuclei stained with H33342; green: ROS stained with H2DCFDA probe. Dose–response curves of cellular ROS treated with tBHP and flavones (e); fluorescence intensity of ROS of microscopy images (f), *** p < 0.001 vs. tBHP; and cell viability induced by tBHP and treated with flavones (g). Full article ">Figure 2
Morphologies of 2D- and 3D-cultured RAW264.7 cells. CLSM and FESEM images of 2D macrophages (a,c); CLSM and SEM images of macrophages cultured on PCL scaffolds (b,d). Blue: cell nuclei stained with H33342; red: F-actin stained with F-actin cytopainter. Full article ">Figure 3
Cytotoxicity and anti-inflammatory activities of 6,3´,4´- and 7,3´,4´-HOFL on 100 ng/mL LPS-induced 2D and 3D RAW264.7 cells. Chemical structures (a,d); cytotoxicity (b,e); and dose–response curves of NO suppression (c,f); n = 6. The inhibitory activity on the gene expressions of IL-1β (g), IL-6 (h), and TNF-α (i) on 2D and 3D cells; n = 9. Inhibitory activities of 6,3´,4´- (50 μM) and 7,3´,4´-HOFL (60 μM) on the protein expressions of iNOS (k), COX-2 (l), and their bands (j); n = 6. The data are shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and lowercase letters represent the differences between the sample groups (p < 0.05). Full article ">Figure 4
6,3´,4´- and 7,3´,4´-HOFL exert anti-inflammatory activity via IL-17, TNF, and JAK-STAT pathways. KEGG enrichment scatter plots of DEGs of LPS vs. Ctrl (a), 6,3´,4´-HOFL vs. LPS (c), and 7,3´,4´-HOFL vs. LPS (d). Clustering heatmap of DEGs involved in IL-17, TNF, and JAK-STAT pathways (b). Full article ">Figure 5
Schematic diagram of the signaling pathways. Created with BioRender.com. Full article ">Figure 6
Verification of gene expressions of key DEGs. Relative mRNA expression of Stat5 (a), Mmp13 (b), Ifnb1 (c), Mmp3 (d), Ccl12 (e), Ccl17 (f), Il2ra (g), Csf2 (h), and Il13ra2 (i). Data are shown as mean ± SD, n = 9, *** p< 0.001. Full article ">Figure 7
The binding target of 6,3´,4´- and 7,3´,4´-HOFL. Computational modeling of 6,3´,4´-HOFL bonded to the ATP-binding pocket on the c-Src (a); the detailed binding scenario of 7,3´,4´-HOFL (b); and dose–response curves of 6,3´,4´- and 7,3´,4´-HOFL on c-Src activity (c). Data are shown as mean ± SD, n = 3. Full article ">

28 pages, 1463 KiB

Open AccessReview

Importance of Insoluble-Bound Phenolics to the Antioxidant Potential Is Dictated by Source Material

by Fereidoon Shahidi and Abul Hossain

Antioxidants 2023, 12(1), 203; https://doi.org/10.3390/antiox12010203 - 15 Jan 2023

Cited by 32 | Viewed by 4549

Abstract

Insoluble-bound phenolics (IBPs) are extensively found in the cell wall and distributed in various tissues/organs of plants, mainly cereals, legumes, and pulses. In particular, IBPs are mainly distributed in the protective tissues, such as seed coat, pericarp, and hull, and are also available [...] Read more.

Insoluble-bound phenolics (IBPs) are extensively found in the cell wall and distributed in various tissues/organs of plants, mainly cereals, legumes, and pulses. In particular, IBPs are mainly distributed in the protective tissues, such as seed coat, pericarp, and hull, and are also available in nutritional tissues, including germ, epicotyl, hypocotyl radicle, and endosperm, among others. IBPs account for 20–60% of the total phenolics in food matrices and can exceed 70% in leaves, flowers, peels, pulps, seeds, and other counterparts of fruits and vegetables, and up to 99% in cereal brans. These phenolics are mostly covalently bound to various macromolecules such as hemicellulose, cellulose, structural protein, arabinoxylan, and pectin, which can be extracted by acid, alkali, or enzymatic hydrolysis along with various thermal and non-thermal treatments. IBPs obtained from various sources exhibited a wide range of biological activities, including antioxidant, anti-inflammatory, antihypertensive, anticancer, anti-obesity, and anti-diabetic properties. In this contribution, the chemistry, distribution, biological activities, metabolism, and extraction methods of IBPs, and how they are affected by various treatments, are summarized. In particular, the effect of thermal and non-thermal processing on the release of IBPs and their antioxidant potential is discussed. Full article

(This article belongs to the Special Issue Soluble and Insoluble-Bound Antioxidants)

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16 pages, 3436 KiB

Open AccessArticle

Potential Antioxidative Activity of Homocysteine in Erythrocytes under Oxidative Stress

by Mingxin Ye, Hui Li, Hongjun Luo, Yongyin Zhou, Wenhong Luo and Zhexuan Lin

Antioxidants 2023, 12(1), 202; https://doi.org/10.3390/antiox12010202 - 15 Jan 2023

Cited by 9 | Viewed by 3156

Abstract

Homocysteine is an amino acid containing a free sulfhydryl group, making it probably contribute to the antioxidative capacity in the body. We recently found that plasma total homocysteine (total-Hcy) concentration increased with time when whole blood samples were kept at room temperature. The [...] Read more.

Homocysteine is an amino acid containing a free sulfhydryl group, making it probably contribute to the antioxidative capacity in the body. We recently found that plasma total homocysteine (total-Hcy) concentration increased with time when whole blood samples were kept at room temperature. The present study was to elucidate how increased plasma total-Hcy is produced and explore the potential physiological role of homocysteine. Erythrocytes and leukocytes were separated and incubated in vitro; the amount of total-Hcy released by these two kinds of cells was then determined by HPLC-MS. The effects of homocysteine and methionine on reactive oxygen species (ROS) production, osmotic fragility, and methemoglobin formation in erythrocytes under oxidative stress were studied. The reducing activities of homocysteine and methionine were tested by ferryl hemoglobin (Hb) decay assay. As a result, it was discovered that erythrocytes metabolized methionine to homocysteine, which was then oxidized within the cells and released to the plasma. Homocysteine and its precursor methionine could significantly decrease Rosup-induced ROS production in erythrocytes and inhibit Rosup-induced erythrocyte’s osmotic fragility increase and methemoglobin formation. Homocysteine (but not methionine) was demonstrated to enhance ferryl Hb reduction. In conclusion, erythrocytes metabolize methionine to homocysteine, which contributes to the antioxidative capability under oxidative stress and might be a supplementary protective factor for erythrocytes against ROS damage. Full article

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Figure 1
(A) The concentrations of plasma total-Hcy after whole blood kept at room temperature for different durations. Plasma total-Hcy content of each whole blood sample (a–f) increased with increasing storage time (0~8 h) at room temperature. (B) Effect of methionine on total-Hcy production from erythrocytes. Addition of 0.01~10 mM methionine led to a concentration-dependent increase of total-Hcy in the supernatant. * p < 0.05 vs. 0 mM methionine group. Full article ">Figure 2
The effects of r-Hcy and methionine on ROS production in Rosup-treated erythrocytes. Incubation of Rosup-treated erythrocytes with 12.5~100 μM r-Hcy (A) and 0.1~5 mM methionine (B) for 0.5~6 h could significantly decrease ROS level in a concentration-dependent manner (n = 6). * p < 0.05 vs. control group (the group without Rosup, r-Hcy, or methionine treatment), & p < 0.05 vs. Rosup group (Rosup treatment alone). Full article ">Figure 3
The protective effects of r-Hcy and methionine on erythrocytes osmotic fragility under oxidative stress. (A) The fragility curve shifted to the right after treatment of erythrocytes with Rosup for 6 h, with 50% hemolysis at 0.43 ± 0.01% NaCl solution. The 0.05~0.8 mM r-Hcy treatment resulted in the fragility curves of Rosup-treated erythrocytes shifted back to the left. (B) The fragility curve shifted to the right after treatment of erythrocytes with Rosup for 6 h, with 50% hemolysis at 0.45 ± 0.03% NaCl solution, and 0.1~2 mM methionine treatment inhibited Rosup-induced erythrocytes osmotic fragility elevation in a dose-dependent manner (n = 4). Full article ">Figure 4
The effects of r-Hcy and methionine on ferryl Hb decay. (A) Representative absorbance spectra taken every 70 s after the addition of 0.8 mM r-Hcy (different color line represented different time point). (B) Absorbance change curves derived from original absorbance spectra in which initial ferryl Hb (Fe4+) spectra were set to zero. Time courses of ferryl (Fe4+) reduction at different concentrations of r-Hcy (C) or methionine (D). Addition of 0.025~0.8 mM r-Hcy to ferryl Hb (Fe4+) solution led to conversion of ferryl Hb (Fe4+) to ferric Hb (Fe3+) in concentration-dependent and time-dependent manners. However, the addition of 0.1~2 mM methionine to ferryl Hb (Fe4+) solution did not significantly change the heme state, compared with control (p > 0.05), except that 0.5 mM methionine caused a slight but significantly increased conversion of ferryl Hb (Fe4+) to ferric Hb (Fe3+). The detailed data can be found in <a href="#app1-antioxidants-12-00202" class="html-app">Supplementary Tables S4 and S5</a>. Full article ">Figure 5
Effects of r-Hcy and methionine on MetHb formation in Rosup-treated erythrocytes (n = 3). The absorbance at 630 nm represented the content of MetHb. (A) r-Hcy (0.8 mM) treatment for 3~9 h significantly inhibited MetHb formation induced by Rosup. (B) Treatment with 0.5~2 mM methionine for 6~9 h significantly decreased formation of MetHb in Rosup-treated erythrocytes. Full article ">Figure 6
The alterations of total-Hcy and GSH levels in erythrocytes after treatment with Rosup in the presence or absence of methionine (n = 3). (A) The concentration of extracellular total-Hcy reduced slightly after Rosup treatment, compared with that of control. Methionine (2 mM) plus Rosup treatment for 3 h or 6 h resulted in a significant increase of extracellular total-Hcy contents, compared with 2 mM methionine or Rosup treatment alone (p < 0.05). (B) Intracellular total-Hcy level did not alter significantly after Rosup treatment (p > 0.05). Methionine (2 mM) plus Rosup treatment for 3 h or 6 h caused a significant increase of intracellular total-Hcy contents, compared with 2 mM methionine or Rosup treatment alone (p < 0.05). (C–F) Rosup treatment significantly decreased intracellular and extracellular reduced GSH levels compared with control (p < 0.05). Methionine treatment alone did not significantly change the intracellular and extracellular reduced GSH and total GSH levels (p > 0.05) but significantly inhibited the decrease of intracellular reduced GSH levels induced by Rosup. * p < 0.05 vs. control group, # p < 0.05 vs. 2 mM Methionine group, & p < 0.05 vs. Rosup group. Full article ">Figure 7
Diagram of erythrocytic-renal cycle and erythrocytic-hepatic cycle. In erythrocytes, methionine is converted to SAM, which is a methyl donor for numerous reactions. After losing its methyl group, SAM becomes SAH, which is then converted to homocysteine. Homocysteine exerts its antioxidative effect and converts to homocystine and then releases to extracellular fluid and transport in circulation to the kidney or liver, which shows enriched expression of BHMT and MTR. In the kidney or liver, homocysteine can be converted back to methionine by the addition of a methyl group. Finally, methionine is synthesized and returned to circulation for further utilization. Full article ">

21 pages, 1056 KiB

Open AccessArticle

Dietary Protected Sodium Butyrate and/or Olive Leaf and Grape-Based By-Product Supplementation Modifies Productive Performance, Antioxidant Status and Meat Quality in Broilers

by Almudena de-Cara, Beatriz Saldaña, Patricia Vázquez and Ana I Rey

Antioxidants 2023, 12(1), 201; https://doi.org/10.3390/antiox12010201 - 15 Jan 2023

Cited by 12 | Viewed by 2746

Abstract

To meet the demand for chicken meat production, new additives that promote growth and health without adverse effects on meat quality are being investigated. This study was conducted to investigate the effect of protected sodium butyrate (PSB) (0 vs. 2 g/kg), an olive [...] Read more.

To meet the demand for chicken meat production, new additives that promote growth and health without adverse effects on meat quality are being investigated. This study was conducted to investigate the effect of protected sodium butyrate (PSB) (0 vs. 2 g/kg), an olive leaf and grape-based by-product (OLG-mix), or a combined supplementation of PSB and OLG-mix on productive performance, antioxidant status, carcass, and meat quality in broilers. PSB improved performance parameters with greater effect in the initial phase. Both, PSB and OLG-mix increased the plasma superoxide dismutase (SOD); however, PSB supplementation was more effective to delay the lipid oxidation of meat from the initial day of storage. OLG-mix produced meat with greater color intensity, b* value and lesser drip losses than PSB. The combination of PSB + OLG-mix did not produce more marked effects that the individual administration; except to control the oxidation of meat. Linear and positive correlations between antioxidant enzymes and weight gain were observed. Significant linear and negative relationships were quantified between plasma SOD and meat lipid oxidation according to dietary treatment. Therefore, the present study would be a first approximation to the possibilities for predicting growth range and meat quality through the evaluation of the blood oxidative status. Full article

(This article belongs to the Special Issue Dietary Antioxidants and Animal Nutrition)

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16 pages, 8566 KiB

Open AccessArticle

Dihydromyricetin Attenuates Diabetic Cardiomyopathy by Inhibiting Oxidative Stress, Inflammation and Necroptosis via Sirtuin 3 Activation

by Yun Chen, Yangyang Zheng, Ruixiang Chen, Jieru Shen, Shuping Zhang, Yunhui Gu, Jiahai Shi and Guoliang Meng

Antioxidants 2023, 12(1), 200; https://doi.org/10.3390/antiox12010200 - 15 Jan 2023

Cited by 18 | Viewed by 2849

Abstract

Dihydromyricetin (DHY), the main flavonoid component in Ampelopsis grossedentata, has important benefits for health. The present study aimed to investigate the exact effects and possible mechanisms of DHY on diabetic cardiomyopathy (DCM). Male C57BL/6 mice and sirtuin 3 (SIRT3) knockout (SIRT3-KO) mice [...] Read more.

Dihydromyricetin (DHY), the main flavonoid component in Ampelopsis grossedentata, has important benefits for health. The present study aimed to investigate the exact effects and possible mechanisms of DHY on diabetic cardiomyopathy (DCM). Male C57BL/6 mice and sirtuin 3 (SIRT3) knockout (SIRT3-KO) mice were injected with streptozotocin (STZ) to induce a diabetic model. Two weeks later, DHY (250 mg/kg) or carboxymethylcellulose (CMC) were administrated once daily by gavage for twelve weeks. We found that DHY alleviated fasting blood glucose (FBG) and triglyceride (TG) as well as glycosylated hemoglobin (HbA1c) levels; increased fasting insulin (FINS); improved cardiac dysfunction; ameliorated myocardial hypertrophy, fibrosis and injury; suppressed oxidative stress, inflammasome and necroptosis; but improved SIRT3 expression in STZ-induced mice. Neonatal rat cardiomyocytes were pre-treated with DHY (80 μM) with or without high glucose (HG) stimulation. The results showed that DHY attenuated cell damage but improved SIRT3 expression and inhibited oxidative stress, inflammasome and necroptosis in cardiomyocytes with high glucose stimulation. Moreover, the above protective effects of DHY on DCM were unavailable in SIRT3-KO mice, implying a promising medical potential of DHY for DCM treatment. In sum, DHY improved cardiac dysfunction; ameliorated myocardial hypertrophy, fibrosis and injury; and suppressed oxidative stress, inflammation and necroptosis via SIRT3 activation in STZ-induced diabetic mice, suggesting DHY may serve as a candidate for an agent to attenuate diabetic cardiomyopathy. Full article

(This article belongs to the Special Issue Antioxidant Therapy for Cardiovascular Diseases)

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Full article ">Figure 1
DHY alleviated FBG, TG and HbA1c levels and increased FINS in STZ-induced C57BL/6 mice. Male C57BL/6 mice (8 weeks old) were intraperitoneally injected with streptozotocin (STZ, 60 mg/kg/day) for 5 consecutive days. Mice in control group were injected with the same amount of citrate buffer. DHY (250 mg/kg) or carboxymethylcellulose (CMC, 0.5%) were administrated 2 weeks later, once daily by gavage for 12 weeks. (A) FBG level. (B) TG level in the serum. (C) HbA1c level in the plasma. (D) FINS level. ** p < 0.01 versus control, # p < 0.05, ## p < 0.01 versus STZ, n = 6. Full article ">Figure 2
DHY improved cardiac dysfunction in STZ-induced C57BL/6 mice. (A) Representative echocardiographs of two-dimensional M-mode and doppler of mice. (B) EF. (C) FS. (D) E/A ratio. ** p < 0.01 versus control; # p < 0.05, ## p < 0.01 versus STZ, n = 6. Full article ">Figure 3
DHY ameliorated myocardial hypertrophy, fibrosis and injury in STZ-induced C57BL/6 mice. (A) The representative images of ventricle tissue with HE staining (bar = 20 μm). (B) Cardiomyocyte areas. (C) Expression of ANP and BNP mRNA. (D–E) The representative images of the myocardium with sirus red staining and Masson’s staining (bar = 20 μm). (F) LDH in the serum. (G) ATP level in the myocardium. ** p < 0.01 versus control, ## p < 0.01 versus STZ, n = 6. Full article ">Figure 4
DHY suppressed oxidative stress, inflammasome and necroptosis but improved SIRT3 expression in STZ-induced C57BL/6 mice. (A) The representative images of myocardium with DHE staining (red) and nuclei with DAPI staining (blue) (bar = 50 μm). (B) The representative images of myocardium with MitoSOX staining (red), mitochondria localization with Mito-tracker staining (green) and nuclei with DAPI staining (blue) (bar = 50 μm). (C–F) Expression of IL-1β, NLRP3, RIPK3 and MLKL protein in the myocardium. (G,H) The representative images of myocardium with TUNEL staining (bar = 20 μm). TUNEL positive cells were quantified. (I) Expression of SIRT3 protein in the myocardium. ** p < 0.01 versus control; # p < 0.05, ## p < 0.01, versus STZ, n = 6. Full article ">Figure 5
DHY attenuated cell damage but improved SIRT3 expression in cardiomyocytes with high glucose stimulation. (A) LDH in the culture medium. (B) ATP level in the cardiomyocytes. (C) Expression of SIRT3 mRNA in the cardiomyocytes. * p < 0.05, ** p < 0.01 versus control; ## p < 0.01 versus HG, n = 6. Full article ">Figure 6
DHY inhibited oxidative stress, inflammasome and necroptosis in cardiomyocytes with high glucose stimulation. (A) The representative images of cardiomyocyte with DHE staining (red) and nuclei with DAPI staining (blue) (bar = 25 μm). (B) The representative images of cardiomyocyte with MitoSOX staining (red), mitochondria localization with Mito-tracker staining (green) and nuclei with DAPI staining (blue) (bar = 50 μm). (C) Caspase 1 was immunofluorescence stained with Alexa Fluor 488-conjugated IgG (green), NLRP3 with Cy3-conjugated IgG (red), and nuclei with DAPI staining (blue) (bar = 25 μm). (D) The representative images of cardiomyocyte with TUNEL staining (green), nuclei with DAPI staining (blue) (bar = 100 μm). TUNEL-positive cells were quantified. (E) Expression of RIPK3 protein. ** p < 0.01 versus control, ## p < 0.01 versus HG, n = 6. Full article ">Figure 7
DHY failed to alleviate diabetic cardiomyopathy in STZ-induced SIRT3-KO mice. Male 8-week-old SIRT3-KO mice were injected with STZ (60 mg/kg/day) for 5 consecutive days. Mice in control group were injected the same amount of citrate buffer. DHY (250 mg/kg) or carboxymethylcellulose (CMC, 0.5%) were administrated 2 weeks later, once daily by gavage for 12 weeks. (A) Representative echocardiographs of two-dimensional M-mode and Doppler of mice. (B) EF. (C) FS. (D) E/A ratio. (E–G) The representative images of the myocardium with HE staining (bar = 100 μm), sirus red staining (bar = 100 μm) and Masson’s staining (bar = 20 μm). (H) Cardiomyocyte areas. (I) Expression of ANP and BNP mRNA. (J) LDH in the serum. (K) ATP level in the myocardium. ** p < 0.01 versus SIRT3-KO. Full article ">

14 pages, 2020 KiB

Open AccessArticle

Obesity Affects Maternal and Neonatal HDL Metabolism and Function

by Julia T. Stadler, Mireille N. M. van Poppel, Christian Wadsack, Michael Holzer, Anja Pammer, David Simmons, David Hill, Gernot Desoye, Gunther Marsche and DALI Core Investigator Group

Antioxidants 2023, 12(1), 199; https://doi.org/10.3390/antiox12010199 - 14 Jan 2023

Cited by 8 | Viewed by 3820

Abstract

Pregravid obesity is one of the major risk factors for pregnancy complications such as gestational diabetes mellitus (GDM) and an increased risk of cardiovascular events in children of affected mothers. However, the biological mechanisms that underpin these adverse outcomes are not well understood. [...] Read more.

Pregravid obesity is one of the major risk factors for pregnancy complications such as gestational diabetes mellitus (GDM) and an increased risk of cardiovascular events in children of affected mothers. However, the biological mechanisms that underpin these adverse outcomes are not well understood. High-density lipoproteins (HDLs) are antiatherogenic by promoting the efflux of cholesterol from macrophages and by suppression of inflammation. Functional impairment of HDLs in obese and GDM-complicated pregnancies may have long-term effects on maternal and offspring health. In the present study, we assessed metrics of HDL function in sera of pregnant women with overweight/obesity of the DALI lifestyle trial (prepregnancy BMI ≥ 29 kg/m²) and women with normal weight (prepregnancy BMI < 25 kg/m²), as well as HDL functionalities in cord blood at delivery. We observed that pregravid obesity was associated with impaired serum antioxidative capacity and lecithin–cholesterol acyltransferase activity in both mothers and offspring, whereas maternal HDL cholesterol efflux capacity was increased. Interestingly, functionalities of maternal and fetal HDL correlated robustly. GDM did not significantly further alter the parameters of HDL function and metabolism in women with obesity, so obesity itself appears to have a major impact on HDL functionality in mothers and their offspring. Full article

(This article belongs to the Special Issue Impact of Antioxidant and Anti-inflammatory Functions of HDL in Diseases)

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Differences between serum lipid levels in normal-weight and overweight/obese pregnant women and in women diagnosed with GDM and its impact on their offspring: (A) shows serum total cholesterol levels, triglycerides, and HDL-C of mothers and (B) of neonates. Data are presented as mean and standard deviation. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Full article ">Figure 2
Differences in serum functionalities in normal-weight and overweight/obese pregnant women and in women with GDM and their offspring. Cholesterol efflux capacity was assessed with a cell-based assay (A,B). The activity of LCAT in mothers (C) and offspring (D) was evaluated. Serum antioxidative capacity was assessed in mothers (E) as well as in paired umbilical cord blood (F). Activity of HDL-associated anti-inflammatory enzyme paraoxonase-1 in mothers (G) and neonates (H) was evaluated. Data are presented as Tukey boxplots showing the median and interquartile ranges as well as minimum and maximum values and outliers. Differences were analyzed by ANOVA or Kruskal–Wallis test based on the distribution of the variable. * p < 0.05, ** p < 0.01. Full article ">Figure 3
Spearman correlation analyses between maternal and cord blood HDL-related parameters and serum functionalities (A) HDL-C, (B) CEC (cholesterol efflux capacity), (C) paraoxonase-1 (PON1) activity, and (D) serum antioxidative capacity. * p < 0.05, ** p < 0.01. Full article ">

15 pages, 5919 KiB

Open AccessArticle

Effects of Thermal Stress on the Antioxidant Capacity, Blood Biochemistry, Intestinal Microbiota and Metabolomic Responses of Luciobarbus capito

by Kun Guo, Rui Zhang, Liang Luo, Shihui Wang, Wei Xu and Zhigang Zhao

Antioxidants 2023, 12(1), 198; https://doi.org/10.3390/antiox12010198 - 14 Jan 2023

Cited by 9 | Viewed by 2572

Abstract

The rise in water temperature caused by global warming is seriously threatening the development of aquatic animals. However, the physiological response mechanism behind the adverse effects of thermal conditions on L. capito remains unclear. In this study, we investigated the physiological responses of [...] Read more.

The rise in water temperature caused by global warming is seriously threatening the development of aquatic animals. However, the physiological response mechanism behind the adverse effects of thermal conditions on L. capito remains unclear. In this study, we investigated the physiological responses of L. capito exposed to thermal stress via biochemical analyses and intestinal microbiota and liver LC–MS metabolomics. The results show that the superoxide dismutase (SOD) and catalase (CAT) activities significantly decrease, while the malondialdehyde (MDA) content, aspartate aminotransferase (AST), acid phosphatase (ACP), alanine aminotransferase (ALT), and albumin (ALB) activities, and glucose (Glu) level significantly increase. Obvious variations in the intestinal microbiota were observed after stress exposure, with increased levels of Proteobacteria and Bacteroidota and decreased levels of Firmicutes, Fusobacteriota, and Actinobacteriota, while levels of several genera of pathogenic bacteria increased. Liver metabolomic analysis showed that stress exposure disturbed metabolic processes, especially of amino acids and lipids. The results of this study indicated that thermal stress caused oxidative stress, disturbed blood biological functioning and intestinal microbiota balance, and damaged amino acids and lipids metabolism of liver in L. capito. Full article

(This article belongs to the Special Issue Antioxidant Defenses in Fish)

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Figure 1

Figure 1
Changes in biochemical indicators in L. capito after thermal stress. Bars indicate mean ± SD (n = 3). (A) Superoxide dismutase (SOD), (B) catalase (CAT), and (C) malondialdehyde (MDA) in liver; (D) aspartate aminotransferase (AST), (E) alanine aminotransferase (ALT), (F) acid phosphatase (ACP), (G) albumin (ALB), and (H) glucose (Glu) in serum. * represents significant difference (p < 0.05), and ** represents highly significant difference (p < 0.01). Full article ">Figure 2
The richness and diversity of intestinal microbial communities in L. capito after thermal stress. (A) Rarefaction curve. (B) Venn diagram. (C) Alpha diversity indices. Bars represent the mean ± SD (n = 5). (D) Beta diversity indicated by PCoA based on Bray–Curtis distance. Full article ">Figure 3
Changes in intestinal microbial composition of L. capito after thermal stress. Relative microbial abundance at the (A) phylum and (B) class level. (C) Heatmap analysis of the top 50 dominant bacterial genera. Red color indicates higher abundance of the genera, and blue color indicates lower abundance, with the intensity of color reflecting the degree of change as indicated in the scale. Full article ">Figure 4
Quality analysis of metabolomics data. The PCA score plot of samples acquired in (A) positive ion and (B) negative ion mode. The OPLS-DA (C) score plot and (D) permutation test for positive ion mode. The OPLS-DA (E) score plot and (F) permutation test for negative ion mode. Full article ">Figure 5
(A) Hierarchical clustering and analysis of the (B) most enriched KEGG pathway of DEMs in the hemolymph of L. capito after thermal stress. Full article ">Figure 6
Significant correlations between intestinal bacteria at the genus level and DEMs. The correlation coefficient is represented by different colors (red, positive correlation; blue, negative correlation) with the intensity reflecting the strength as indicated in the scale. * represents significantly negative or positive correlation (* p < 0.05; ** p < 0.01; *** p < 0.001). Full article ">

17 pages, 2467 KiB

Open AccessArticle

Oleuropein Attenuates Oxidative Stress in Human Trophoblast Cells

by Andrea Pirković, Aleksandra Vilotić, Sunčica Borozan, Mirjana Nacka-Aleksić, Žanka Bojić-Trbojević, Milica Jovanović Krivokuća, Maurizio Battino, Francesca Giampieri and Dragana Dekanski

Antioxidants 2023, 12(1), 197; https://doi.org/10.3390/antiox12010197 - 14 Jan 2023

Cited by 9 | Viewed by 2819

Abstract

Olive-derived bioactive compound oleuropein was evaluated against damage induced by hydrogen peroxide in human trophoblast cells in vitro, by examining the changes in several markers implicated in oxidative stress interactions in the placenta. Trophoblast HTR-8/SVneo cells were preincubated with OLE at 10 [...] Read more.

Olive-derived bioactive compound oleuropein was evaluated against damage induced by hydrogen peroxide in human trophoblast cells in vitro, by examining the changes in several markers implicated in oxidative stress interactions in the placenta. Trophoblast HTR-8/SVneo cells were preincubated with OLE at 10 and 100 µM and exposed to H₂O₂, as a model of oxidative stress. Protein and lipid peroxidation, as well as antioxidant enzymes’ activity, were determined spectrophotometrically, and DNA damage was evaluated by comet assay. iNOS protein expression was assessed by Western blot, while the mRNA expression of pro- and anti-apoptotic genes BAX and BCL2 and transcription factor NFE2L2, as well as cytokines IL-6 and TNF α were determined by qPCR. Oleuropein demonstrated cytoprotective effects against H₂O₂ in trophoblast cells by significantly improving the antioxidant status and preventing protein and lipid damage, as well as reducing the iNOS levels. OLE reduced the mRNA expression of IL-6 and TNF α, however, it did not influence the expression of NFE2L2 or the BAX/BCL2 ratio after H₂O₂ exposure. Oleuropein per se did not lead to any adverse effects in HTR-8/SVneo cells under the described conditions, confirming its safety in vitro. In conclusion, it significantly attenuated oxidative damage and restored antioxidant functioning, confirming its protective role in trophoblast. Full article

(This article belongs to the Special Issue Oxidative Stress in Reproduction)

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Figure 1
Cytotoxicity of oleuropein (OLE) in HTR-8/SVneo cells after 24 h treatment with a range of concentrations (1, 10, 25, 50, 100 and 200 µM). Full article ">Figure 2
Cytoprotective effect of oleuropein (OLE) on H2O2-induced damage in HTR-8/SVneo cells. The data are expressed as mean + SEM. ** p < 0.01, **** p < 0.0001. Full article ">Figure 3
(A) Genotoxic potential of oleuropein (OLE) and antigenotoxic effects against H2O2-induced damage in HTR-8/SVneo cells. The data are expressed as mean + SEM. **** p < 0.0001. (B) Appearance of comets without DNA damage (Non damaged) and with DNA damage (Damaged). Full article ">Figure 4
The effect of oleuropein (OLE) on activity of antioxidant enzymes in H2O2-exposed HTR-8/SVneo cells. The investigated parameters include catalase (CAT) activity (A), gluthatione peroxidase (GPx) activity (B) and superoxide dismutase (SOD) activity (C). The data are expressed as mean + SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Full article ">Figure 5
The effect of oleuropein (OLE) on lipid peroxidation and protein damage in H2O2-exposed HTR-8/SVneo cells. The investigated parameters include malondialdehyde (MDA) concentration (A), extracellular lactate dehydrogenase (LDH) activity (B), reduced glutathione (GSH) concentration (C), protein carbonyl group (CG) concentration (D) and nitrite (NO2−) concentration (E). The data are expressed as mean + SEM., *** p < 0.001, **** p < 0.0001. Full article ">Figure 6
The effect of oleuropein (OLE) on the inducible nitric oxide synthase (iNOS) protein expression in H2O2-exposed HTR-8/SVneo cells. Upper panel, representative Western blot of iNOS; lower panel, densitometric analysis of iNOS expression in HTR-8/SVneo cells. The data are expressed as mean + SEM. ** p < 0.01, **** p < 0.0001. Full article ">Figure 7
The effect of oleuropein (OLE) on the mRNA expression of apoptotic marker BAX, the anti-apoptotic marker BCL2, the mRNA ratio of BAX to BCL2 and the mRNA expression of redox-regulated transcription factor NFE2L2, interleukin 6 (IL6) and tumor necrosis factor α (TNFα), in H2O2-exposed HTR-8/SVneo cells. The data are expressed as mean + SEM. ** p < 0.01. Full article ">

18 pages, 949 KiB

Open AccessReview

Thermogenic Adipose Redox Mechanisms: Potential Targets for Metabolic Disease Therapies

by Ashley K. Putman, G. Andres Contreras and Emilio P. Mottillo

Antioxidants 2023, 12(1), 196; https://doi.org/10.3390/antiox12010196 - 14 Jan 2023

Cited by 4 | Viewed by 2909

Abstract

Metabolic diseases, such as diabetes and non-alcoholic fatty liver disease (NAFLD), have several negative health outcomes on affected humans. Dysregulated energy metabolism is a key component underlying the pathophysiology of these conditions. Adipose tissue is a fundamental regulator of energy homeostasis that utilizes [...] Read more.

Metabolic diseases, such as diabetes and non-alcoholic fatty liver disease (NAFLD), have several negative health outcomes on affected humans. Dysregulated energy metabolism is a key component underlying the pathophysiology of these conditions. Adipose tissue is a fundamental regulator of energy homeostasis that utilizes several redox reactions to carry out the metabolism. Brown and beige adipose tissues, in particular, perform highly oxidative reactions during non-shivering thermogenesis to dissipate energy as heat. The appropriate regulation of energy metabolism then requires coordinated antioxidant mechanisms to counterbalance the oxidation reactions. Indeed, non-shivering thermogenesis activation can cause striking changes in concentrations of both oxidants and antioxidants in order to adapt to various oxidative environments. Current therapeutic options for metabolic diseases either translate poorly from rodent models to humans (in part due to the challenges of creating a physiologically relevant rodent model) or tend to have numerous side effects, necessitating novel therapies. As increased brown adipose tissue activity results in enhanced energy expenditure and is associated with beneficial effects on metabolic health, such as decreased obesity, it has gathered great interest as a modulator of metabolic disease. One potential reason for the beneficial health effects may be that although non-shivering thermogenesis is enormously oxidative, it is also associated with decreased oxidant formation after its activation. However, targeting its redox mechanisms specifically to alter metabolic disease remains an underexplored area. Therefore, this review will discuss the role of adipose tissue in energy homeostasis, non-shivering thermogenesis in adults, and redox mechanisms that may serve as novel therapeutic targets of metabolic disease. Full article

(This article belongs to the Special Issue From Nature to Synthetic Small Molecule Antioxidants: New Candidates in Drug Discovery)

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15 pages, 1140 KiB

Open AccessReview

Oxidative Stress and Antioxidants in Chronic Rhinosinusitis with Nasal Polyps

by Junhu Tai, Jae-Min Shin, Jaehyung Park, Munsoo Han and Tae Hoon Kim

Antioxidants 2023, 12(1), 195; https://doi.org/10.3390/antiox12010195 - 14 Jan 2023

Cited by 9 | Viewed by 3289

Abstract

Oxidative stress results from an imbalance between the production of reactive oxygen species and the body’s antioxidant defense system. It plays an important role in the regulation of the immune response and can be a pathogenic factor in various diseases. Chronic rhinosinusitis (CRS) [...] Read more.

Oxidative stress results from an imbalance between the production of reactive oxygen species and the body’s antioxidant defense system. It plays an important role in the regulation of the immune response and can be a pathogenic factor in various diseases. Chronic rhinosinusitis (CRS) is a complex and heterogeneous disease with various phenotypes and endotypes. Recently, an increasing number of studies have proposed that oxidative stress (caused by both environmental and intrinsic stimuli) plays an important role in the pathogenesis and persistence of CRS. This has attracted the attention of several researchers. The relationship between the presence of reactive oxygen species composed of free radicals and nasal polyp pathology is a key topic receiving attention. This article reviews the role of oxidative stress in respiratory diseases, particularly CRS, and introduces potential therapeutic antioxidants that may offer targeted treatment for CRS. Full article

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13 pages, 3533 KiB

Open AccessArticle

Nootkatone Supplementation Ameliorates Carbon Tetrachloride-Induced Acute Liver Injury via the Inhibition of Oxidative Stress, NF-κB Pathways, and the Activation of Nrf2/HO-1 Pathway

by Chongshan Dai, Xueyong Zhang, Jiahao Lin and Jianzhong Shen

Antioxidants 2023, 12(1), 194; https://doi.org/10.3390/antiox12010194 - 13 Jan 2023

Cited by 16 | Viewed by 2782

Abstract

Acute liver injury is a type of liver diseases, and it has raised concerns worldwide due to the lack of effective therapies. The aim of this study is to investigate the protective effects of nootkatone (NOOT) on carbon tetrachloride (CCl₄)-caused acute [...] Read more.

Acute liver injury is a type of liver diseases, and it has raised concerns worldwide due to the lack of effective therapies. The aim of this study is to investigate the protective effects of nootkatone (NOOT) on carbon tetrachloride (CCl₄)-caused acute liver injury in mice. Mice were randomly divided into control, CCl₄ model, NOOT, and NOOT (5, 10, and 20 mg/kg/day) plus CCl₄ groups, respectively. Mice in the CCl₄ plus NOOT groups were orally administrated with NOOT at 5, 10, and 20 mg/kg/days for seven days prior to 0.3% CCl₄ injection at 10 mL/kg body weight, respectively. Our results showed that NOOT supplementation significantly ameliorated CCl₄-induced increases of serum AST and ALT levels, hepatocyte necrosis, inflammatory response, oxidative stress, and caspases-9 and -3 activities in the livers of mice. Moreover, NOOT supplementation significantly upregulated the expression of Nrf2 and HO-1 mRNAs but downregulated the expression of NF-κB mRNAs and the levels of IL-1β, IL-6, and TNF-α proteins in the liver tissues, compared to those in the CCl₄ model group. In conclusion, for the first time, our results reveal that NOOT could offer protective effects against CCl₄-caused oxidative stress and inflammatory response via the opposite regulation of Nrf2/HO-1 pathway and NF-κB pathway. Full article

(This article belongs to the Special Issue Oxidative Stress as a Crucial Factor in Liver Diseases: From Basic Research to Clinical Trials)

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19 pages, 3253 KiB

Open AccessArticle

Development of Dietary Thiol Antioxidant via Reductive Modification of Whey Protein and Its Application in the Treatment of Ischemic Kidney Injury

by Yang Sui, Rui Jiang, Manabu Niimi, Jingru Hong, Qiaojing Yan, Zhuheng Shi and Jian Yao

Antioxidants 2023, 12(1), 193; https://doi.org/10.3390/antiox12010193 - 13 Jan 2023

Cited by 3 | Viewed by 2731

Abstract

Thiol antioxidants play important roles in cell and body defense against oxidative stress. In body fluid, albumin is the richest source of thiol antioxidants. One recent study showed that the reductive modification of thiol residues in albumin potentiated its antioxidative activity. Given that [...] Read more.

Thiol antioxidants play important roles in cell and body defense against oxidative stress. In body fluid, albumin is the richest source of thiol antioxidants. One recent study showed that the reductive modification of thiol residues in albumin potentiated its antioxidative activity. Given that whey protein (WP) contains albumin and other thiol-active proteins, this property of WP could be exploited to develop novel thiol antioxidants. The aim of this study was to address this possibility. WP was reductively modified with dithiothreitol (DTT). The modified protein exhibited significantly elevated free sulfhydryl groups (-SH) and thiol antioxidative activity. It detoxified H₂O₂ and prevented H₂O₂-initiated protein oxidation and cell death in a -SH group-dependent way in vitro. In addition, it reacted with GSH/GSSG and altered the GSH/GSSG ratio via thiol–disulfide exchange. In vivo, oral administration of the reductively modified WP prevented oxidative stress and renal damage in a mouse model of renal injury caused by ischemia reperfusion. It significantly improved renal function, oxidation, inflammation, and cell injury. These protective effects were not observed in the WP control and were lost after blocking the -SH groups with maleimide. Furthermore, albumin, one of the ingredients of WP, also exhibited similar protective effects when reductively modified. In conclusion, the reductive modification of thiol residues in WP transformed it into a potent thiol antioxidant that protected kidneys from ischemia reperfusion injury. Given that oxidative stress underlies many life-threatening diseases, the reductively modified dietary protein could be used for the prevention and treatment of many oxidative-stress-related conditions, such as cardiovascular diseases, cancer, and aging. Full article

(This article belongs to the Special Issue Dietary Supplements and Oxidative Stress)

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15 pages, 3861 KiB

Open AccessArticle

The Interaction between Oxidative Stress Biomarkers and Gut Microbiota in the Antioxidant Effects of Extracts from Sonchus brachyotus DC. in Oxazolone-Induced Intestinal Oxidative Stress in Adult Zebrafish

by Juan Yang, Wei-Wei Zhou, Dong-Dong Shi, Fang-Fang Pan, Wen-Wen Sun, Pei-Long Yang and Xiu-Mei Li

Antioxidants 2023, 12(1), 192; https://doi.org/10.3390/antiox12010192 - 13 Jan 2023

Cited by 9 | Viewed by 3178

Abstract

Oxidative stress is a phenomenon caused by an imbalance between the production and accumulation of reactive oxygen species in cells and tissues that eventually leads to the production of various diseases. Here, we investigated the antioxidant effects of the extract from Sonchus brachyotus [...] Read more.

Oxidative stress is a phenomenon caused by an imbalance between the production and accumulation of reactive oxygen species in cells and tissues that eventually leads to the production of various diseases. Here, we investigated the antioxidant effects of the extract from Sonchus brachyotus DC. (SBE) based on the 0.2% oxazolone-induced intestinal oxidative stress model of zebrafish. Compared to the model group, the treatment group alleviated oxazolone-induced intestinal tissue damage and reduced the contents of malondialdehyde, reactive oxygen species, IL-1β, and TNF-α and then increased the contents of superoxide dismutase, glutathione peroxidase, and IL-10. The 16s rDNA gene sequencing findings demonstrated that SBE could increase the relative abundance of Fusobacteriota, Actinobacteriota, and Firmicutes and decrease the relative abundance of Proteobacteria. Based on the correlation analysis between the oxidative stress biomarkers and intestinal flora, we found that the trends of oxidative stress biomarkers were significantly correlated with intestinal microorganisms, especially at the genus level. The correlations of MDA, IL-1β, and TNF-α were significantly negative with Shewanella, while SOD, GSH-Px, and IL-10 were significantly positive with Cetobacterium, Gemmobacter, and Flavobacterium. Consequently, we concluded that the antioxidant effect of SBE was realized through the interaction between oxidative stress biomarkers and gut microbiota. Full article

(This article belongs to the Topic Antioxidant Activity of Natural Products)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
Changes in ROS content and MDA content caused by oxazolone injection time. The values are expressed as the mean ± SD. (n = 6 per group). Statistical differences between groups during the indicated time course were obtained according to repeated-measurement ANOVA (compared with the CON group, # p < 0.05, ## p < 0.01). Full article ">Figure 2
Effects of SBE on the intestinal tissue of zebrafish. Representative hematoxylin and eosin (H&E)-stained intestinal sections and semiquantitative scoring showed tissue damage. The values are expressed as the mean ± SD. (n = 3). The thin black arrows represent degeneration, necrosis, and detachment of mucosal epithelial cells. The hollow arrows represent the exposed muscle layer. The folded arrow represents neutrophil infiltration. Thick black arrows represent villi breakage. Groups with different letters statistically differ (compared with the CON group, # p < 0.05, ## p < 0.01). Full article ">Figure 3
Effects of SBE on the oxidative stress biomarkers. Zebrafish intestines were collected, and the ROS relative intensity of fluorescence (A), the MDA contents (B), and the enzyme activities of SOD (C), GSH-Px (D), and CAT (E) are shown. Fold changes are expressed as means ± SD (n = 30 per group). Groups with different letters statistically differ (compared with the CON group, # p < 0.05, ## p < 0.01; compared with the Oxa group, * p < 0.05, ** p < 0.01). Full article ">Figure 4
Effects of SBE on the inflammatory factors. Zebrafish intestines were collected, and quantified real-time PCR and mRNA expressions of TNF-α (A), IL-1β (B), and IL-10 (C) are shown. Fold changes are expressed as means ± SD (n = 30 per group). Groups with different letters statistically differ (compared with the CON group, ## p < 0.01; compared with the Oxa group, ** p < 0.01). Full article ">Figure 5
Effects of SBE on intestinal microbiota in zebrafish. A Venn diagram shows the overlap of the OTUs identified in the intestinal microbiota among CON, Oxa, and SBE groups (A). Analysis of alpha diversity (Shannon index) was used to detect differences between the groups (B). Plots of unweighted UniFrac-based PCoA (C). The top 15 bacteria, with a maximum abundance of gut bacteria at the phylum level (D). The top 15 bacteria, with a maximum abundance of gut bacteria at the genus level (E). Full article ">Figure 6
The correlation analysis between the contents of oxidative stress biomarkers and intestinal flora. The variation of oxidative stress biomarkers with relation to the intestinal microbiota. Correlation between biomarkers and intestinal microbiota at the phylum level (A). Histogram about the abundance of microbe that correlated with biomarkers contents at the phylum level (B–D). Correlation between biomarkers and intestinal microbiota at the genus level (E). Histogram about the abundance of microbe that correlated with biomarkers contents at the genus level (F–I). Mechanistic diagram of SBE regulating oxidative stress biomarkers via intestinal flora in zebrafish (J). A redder color indicates a positive correlation, while a bluer color indicates a negative correlation (* p < 0.05, ** p < 0.01. n ≥ 3/per group). Full article ">Figure 7
The correlation analysis between intestinal inflammatory factors and intestinal flora. Correlation between the mRNA expressions of TNF-α, IL-1β, and IL-10 and intestinal microbiota at the phylum level (A). Histogram about the abundance of microbe that significantly correlated with the expression of IL-10 at the phylum level (B,C). Correlation between the mRNA expressions of TNF-α, IL-1β, and IL-10 and intestinal microbiota at the genus level (D). Histogram about the abundance of microbe that significantly correlated with the expression of TNF-α, IL-1β, and IL-10 at the genus level (E–H). Mechanistic diagram of SBE regulating inflammatory factors via intestinal flora in zebrafish (I). A redder color indicates a positive correlation, while a bluer color indicates a negative correlation (* p < 0.05, ** p < 0.01. n ≥ 3/per group). Full article ">

15 pages, 3070 KiB

Open AccessArticle

Effect of Encapsulation Material on Lipid Bioaccessibility and Oxidation during In Vitro Digestion of Black Seed Oil

by Jon Alberdi-Cedeño, Martha Aichner, Agnes Mistlberger-Reiner, Aimin Shi and Marc Pignitter

Antioxidants 2023, 12(1), 191; https://doi.org/10.3390/antiox12010191 - 13 Jan 2023

Cited by 2 | Viewed by 2044

Abstract

Different encapsulation materials might not only affect lipid hydrolysis but also lipid oxidation during in vitro digestion. Thus, this study aimed to investigate the effect of two commonly used shell materials, starch and gelatin, on the extent of lipolysis and bioaccessibility of the [...] Read more.

Different encapsulation materials might not only affect lipid hydrolysis but also lipid oxidation during in vitro digestion. Thus, this study aimed to investigate the effect of two commonly used shell materials, starch and gelatin, on the extent of lipolysis and bioaccessibility of the main and some minor lipid compounds, as well as on the oxidative status in encapsulated black seed oil (Nigella sativa) during in vitro digestion. The study was carried out using ¹H nuclear magnetic resonance spectroscopy, liquid chromatography-mass spectrometry and high-performance liquid chromatography-UV. It was shown that starch increased the level of lipid hydrolysis in black seed oil during gastric in vitro digestion, while no differences were observed in the intestinal digestates between starch-encapsulated oil and gelatin-encapsulated oil. Similarly, the bioaccessibility of minor compounds (tocopherols, sterols and thymoquinone) was not influenced by the shell materials. However, regarding lipid oxidation, a 20- and 10-fold rise of free oxylipins was obtained in oils encapsulated by starch and gelatin, respectively, after intestinal in vitro digestion. This study evidenced that gelatin rather than starch should be used for the encapsulation of oils to minimize the digestion-induced formation of bioactive oxylipins. Full article

(This article belongs to the Special Issue Antioxidants and Oxidative Stability in Fats and Oils)

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Particle concentration (particles/mL) (A) and median particle size (nm) (B) in the digestates of starch-encapsulated black seed oil (BS1) or gelatin-encapsulated black seed oil (BS2) after oral (O-BS), gastric (G-BS), or intestinal (I-BS) in vitro digestion. Particle size distribution in the different digestates of BS1 (C) and BS2 (D), respectively. Different letters indicate statistically significant differences among the samples analyzed by one-way ANOVA (p < 0.05). Asterisks indicate statistically significant differences between the indicated samples analyzed by student’s t-test (*** = p < 0.001, * = p < 0.05). Full article ">Figure 2
Molar percentage of linoleic, oleic and saturated fatty acids (FA) plus acyl groups (AG) (FA+AG) in relation to the total moles of all kinds of AG and FA (FA+AG) in black seed oils 1 and 2 (BS1 and BS2) and in the digestates of these oils after gastric (G-BS1 and G-BS2) and intestinal (I-BS1 and I-BS2) digestion. Different letters indicate statistically significant differences among the samples analyzed by one-way ANOVA (p < 0.05). Full article ">Figure 3
Concentration of n-alkanals expressed as mmol per mol of AG+FA in black seed oils 1 and 2 (BS1 and BS2) and in the digestates of these oils after gastric (G-BS1 and G-BS2) and intestinal (I-BS1 and I-BS2) in vitro digestion. Different letters indicate statistically significant differences among the samples analyzed by one-way ANOVA (p < 0.05). Full article ">

19 pages, 7523 KiB

Open AccessArticle

Cerium Oxide Nanoparticles Conjugated with Tannic Acid Prevent UVB-Induced Oxidative Stress in Fibroblasts: Evidence of a Promising Anti-Photodamage Agent

by Regina G. Daré, Elayaraja Kolanthai, Craig J. Neal, Yifei Fu, Sudipta Seal, Celso V. Nakamura and Sueli O. S. Lautenschlager

Antioxidants 2023, 12(1), 190; https://doi.org/10.3390/antiox12010190 - 12 Jan 2023

Cited by 21 | Viewed by 3480

Abstract

Exposure to ultraviolet radiation induces photodamage towards cellular macromolecules that can progress to photoaging and photocarcinogenesis. The topical administration of compounds that maintain the redox balance in cells presents an alternative approach to combat skin oxidative damage. Cerium oxide nanoparticles (CNPs) can act [...] Read more.

Exposure to ultraviolet radiation induces photodamage towards cellular macromolecules that can progress to photoaging and photocarcinogenesis. The topical administration of compounds that maintain the redox balance in cells presents an alternative approach to combat skin oxidative damage. Cerium oxide nanoparticles (CNPs) can act as antioxidants due to their enzyme-like activity. In addition, a recent study from our group has demonstrated the photoprotective potential of tannic acid (TA). Therefore, this work aimed to synthesize CNPs associated with TA (CNP-TA) and investigate its photoprotective activity in L929 fibroblasts exposed to UVB radiation. CNP conjugation with TA was confirmed by UV–Vis spectra and X-ray photoelectron spectroscopy. Bare CNPs and CNP-TA exhibited particle sizes of ~5 and ~10 nm, superoxide dismutase activity of 3724 and 2021 unit/mg, and a zeta potential of 23 and −19 mV, respectively. CNP-TA showed lower cytotoxicity than free TA and the capacity to reduce the oxidative stress caused by UVB; supported by the scavenging of reactive oxygen species, the prevention of endogenous antioxidant system depletion, and the reduction in oxidative damage in lipids and DNA. Additionally, CNP-TA improved cell proliferation and decreased TGF-β, metalloproteinase-1, and cyclooxygenase-2. Based on these results, CNP-TA shows therapeutic potential for protection against photodamage, decreasing molecular markers of photoaging and UVB-induced inflammation. Full article

(This article belongs to the Special Issue Nanoparticles with Antioxidant Activity)

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33 pages, 4674 KiB

Open AccessFeature PaperArticle

Antioxidant Activity and Phenolic Compound Identification and Quantification in Western Australian Honeys

by Ivan Lozada Lawag, Md Khairul Islam, Tomislav Sostaric, Lee Yong Lim, Katherine Hammer and Cornelia Locher

Antioxidants 2023, 12(1), 189; https://doi.org/10.3390/antiox12010189 - 12 Jan 2023

Cited by 24 | Viewed by 4741

Abstract

This study reports on the total phenolic content and antioxidant activity as well as the phenolic compounds that are present in Calothamnus spp. (Red Bell), Agonis flexuosa (Coastal Peppermint), Corymbia calophylla (Marri) and Eucalyptus marginata (Jarrah) honeys from Western Australia. The honey’s total [...] Read more.

This study reports on the total phenolic content and antioxidant activity as well as the phenolic compounds that are present in Calothamnus spp. (Red Bell), Agonis flexuosa (Coastal Peppermint), Corymbia calophylla (Marri) and Eucalyptus marginata (Jarrah) honeys from Western Australia. The honey’s total phenolic content (TPC) was determined using a modified Folin–Ciocalteu assay, while their total antioxidant activity was determined using FRAP and DPPH assays. Phenolic constituents were identified using a High Performance Thin-Layer Chromatography (HTPLC)-derived phenolic database, and the identified phenolic compounds were quantified using HPTLC. Finally, constituents that contribute to the honeys’ antioxidant activity were identified using a DPPH-HPTLC bioautography assay. Based on the results, Calothamnus spp. honey (n = 8) was found to contain the highest (59.4 ± 7.91 mg GAE/100 g) TPC, followed by Eucalyptus marginata honey (50.58 ± 3.76 mg GAE/100 g), Agonis flexuosa honey (36.08 ± 4.2 mg GAE/100 g) and Corymbia calophylla honey (29.15 ± 5.46 mg GAE/100 g). In the FRAP assay, Calothamnus spp. honey also had the highest activity (9.24 ± 1.68 mmol Fe²⁺/kg), followed by Eucalyptus marginata honey (mmol Fe²⁺/kg), whereas Agonis flexuosa (5.45 ± 1.64 mmol Fe²⁺/kg) and Corymbia calophylla honeys (4.48 ± 0.82 mmol Fe²⁺/kg) had comparable FRAP activity. In the DPPH assay, when the mean values were compared, it was found that Calothamnus spp. honey again had the highest activity (3.88 ± 0.96 mmol TE/kg) while the mean DPPH antioxidant activity of Eucalyptus marginata, Agonis flexuosa, and Corymbia calophylla honeys were comparable. Kojic acid and epigallocatechin gallate were found in all honeys, whilst other constituents (e.g., m-coumaric acid, lumichrome, gallic acid, taxifolin, luteolin, epicatechin, hesperitin, eudesmic acid, syringic acid, protocatechuic acid, t-cinnamic acid, o-anisic acid) were only identified in some of the honeys. DPPH-HPTLC bioautography demonstrated that most of the identified compounds possess antioxidant activity, except for t-cinnamic acid, eudesmic acid, o-anisic acid, and lumichrome. Full article

(This article belongs to the Special Issue Antioxidant Activity of Honey Bee Products)

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25 pages, 1552 KiB

Open AccessArticle

In Mild and Moderate Acute Ischemic Stroke, Increased Lipid Peroxidation and Lowered Antioxidant Defenses Are Strongly Associated with Disabilities and Final Stroke Core Volume

by Michael Maes, Francis F. Brinholi, Ana Paula Michelin, Andressa K. Matsumoto, Laura de Oliveira Semeão, Abbas F. Almulla, Thitiporn Supasitthumrong, Chavit Tunvirachaisakul and Decio S. Barbosa

Antioxidants 2023, 12(1), 188; https://doi.org/10.3390/antiox12010188 - 12 Jan 2023

Cited by 4 | Viewed by 2582

Abstract

In acute ischemic stroke (AIS), there are no data on whether oxidative stress biomarkers have effects above and beyond known risk factors and measurements of stroke volume. This study was conducted in 122 mild-moderate AIS patients and 40 controls and assessed the modified [...] Read more.

In acute ischemic stroke (AIS), there are no data on whether oxidative stress biomarkers have effects above and beyond known risk factors and measurements of stroke volume. This study was conducted in 122 mild-moderate AIS patients and 40 controls and assessed the modified ranking scale (mRS) at baseline, and 3 and 6 months later. We measured lipid hydroperoxides (LOOH), malondialdehyde (MDA), advanced oxidation protein products, paraoxonase 1 (PON1) activities and PON1 Q192R genotypes, high density lipoprotein cholesterol (HDL), sulfhydryl (-SH) groups), and diffusion-weighted imaging (DWI) stroke volume and fluid-attenuated inversion recovery (FLAIR) signal intensity. We found that (a) AIS is characterized by lower chloromethyl acetate CMPAase PON1 activity, HDL and -SH groups and increased LOOH and neurotoxicity (a composite of LOOH, inflammatory markers and glycated hemoglobin); (b) oxidative and antioxidant biomarkers strongly and independently predict mRS scores 3 and 6 months later, DWI stroke volume and FLAIR signal intensity; and (c) the PON1 Q192R variant has multiple effects on stroke outcomes that are mediated by its effects on antioxidant defenses and lipid peroxidation. Lipid peroxidation and lowered -SH and PON1-HDL activity are drug targets to prevent AIS and consequent neurodegenerative processes and increased oxidative reperfusion mediators due to ischemia-reperfusion injury. Full article

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Importance chart of a neural network analysis with acute ischemic stroke (AIS) versus controls as input variables. LOOH: lipid hydroperoxides; zCMPAase: z transformation of chloromethyl acetate (CMPA)ase; zAREase: z transformation of aryl esterase; WBC: white blood cells; -SH: sulfhydryl; NLR: neutrophil/lymphocyte ratio; zHDL: z transformation of high-density lipoprotein cholesterol; HbA1c: glycated hemoglobin; CRP: C-reactive protein; Castelli 1: an index of the Castelli risk index 1; PON: paraoxonase; NOx: nitric oxide metabolites; AOPP: advanced oxidation protein products; AIP: and index of the atherogenic index of plasma; MDA: malondialdehyde. Full article ">Figure 2
Importance chart of a neural network analysis with acute ischemic stroke (AIS) versus controls as input variables and composite scores of the predictors as input variables. zNT—z composite score of the neurotoxic analytes assayed in our study; HT—hypertension; zANTIOX—z composite score of the antioxidants assayed in our study; Castelli 1—an index of the Castelli risk index 1; BMI—body mass index; PON—paraoxonase; AIP—and index of the atherogenic index of plasma. Full article ">Figure 3
Partial regression of the baseline National Institutes of Health Stroke Scale (NIHSS) score on the neurotoxicity (zNT) index (after adjusting for age, sex, BMI, and smoking). Full article ">Figure 4
Partial regression of the baseline modified Rankin score (mRS) on an index of antioxidant defences (after adjusting for age, sex, BMI, and smoking). Full article ">Figure 5
Partial regression of the modified Rankin score (mRS) at 3 months on the neurotoxicity index (zNT) after adjusting for age, sex, BMI, and smoking. Full article ">Figure 6
Results of partial least squares (PLS) analysis with severity of acute ischemic stroke (AIS) as final outcome variable. The direct explanatory variables are the neurotoxicity (zNT) index, white blood cell (WBC) count, antioxidant defenses (zANTIOX), classical risk factors (RISK), paraoxonase 1 (PON1) status, including chloromethyl phenylacetate CMPAase and arylesterase (ARE)ase activity and the PON1 Q192 genotype (entered as an additive model coding RR as 2, QR as 1 and QQ 0), high density lipoprotein cholesterol (HDL), and the CMPAase + HDL complex. zNT—composite based on C-reactive protein, neutrophil/lymphocyte ratio (NLR), lipid hydroperoxides (LOOH) and glycated hemoglobin (HbA1c). zANTIOX—a composte based on HDL, CMPAse and sulfhydryl (-SH) groups. AIS was conceptualized as a reflective model with the modified Rankin score (mRS) and National Institutes of Health Stroke Scale (NIHSS) scores as well as an ordinal variable based on the diagnostic groups shown in <a href="#antioxidants-12-00188-t001" class="html-table">Table 1</a> as manifestations. DysLD—dyslipidemia, HT—hypertension, PrStroke—prior stroke. Full article ">

27 pages, 472 KiB

Open AccessReview

The Therapeutic Alliance between Pomegranate and Health Emphasizing on Anticancer Properties

by Panagiota D. Pantiora, Alexandros I. Balaouras, Ioanna K. Mina, Christoforos I. Freris, Athanasios C. Pappas, Georgios P. Danezis, Evangelos Zoidis and Constantinos A. Georgiou

Antioxidants 2023, 12(1), 187; https://doi.org/10.3390/antiox12010187 - 12 Jan 2023

Cited by 13 | Viewed by 4331

Abstract

Pomegranate is a fruit bearing-plant that is well known for its medicinal properties. Pomegranate is a good source of phenolic acids, tannins, and flavonoids. Pomegranate juice and by-products have attracted the scientific interest due to their potential health benefits. Currently, the medical community [...] Read more.

Pomegranate is a fruit bearing-plant that is well known for its medicinal properties. Pomegranate is a good source of phenolic acids, tannins, and flavonoids. Pomegranate juice and by-products have attracted the scientific interest due to their potential health benefits. Currently, the medical community has showed great interest in exploiting pomegranate potential as a protective agent against several human diseases including cancer. This is demonstrated by the fact that there are more than 800 reports in the literature reporting pomegranate’s anticancer properties. This review is an update on the research outcomes of pomegranate’s potential against different types of human diseases, emphasizing on cancer. In addition, perspectives of potential applications of pomegranate, as a natural additive aiming to improve the quality of animal products, are discussed. Full article

(This article belongs to the Special Issue Natural Products: Biological-, Antioxidant Properties and Health Effects - 2nd Edition)

18 pages, 4113 KiB

Open AccessArticle

Identification of Lipocalin 2 as a Ferroptosis-Related Key Gene Associated with Hypoxic-Ischemic Brain Damage via STAT3/NF-κB Signaling Pathway

by Lianxiang Luo, Liyan Deng, Yongtong Chen, Rui Ding and Xiaoling Li

Antioxidants 2023, 12(1), 186; https://doi.org/10.3390/antiox12010186 - 12 Jan 2023

Cited by 28 | Viewed by 5024

Abstract

Hypoxic-ischemic brain damage (HIBD) is a common cause of death or mental retardation in newborns. Ferroptosis is a novel form of iron-dependent cell death driven by lipid peroxidation, and recent studies have confirmed that ferroptosis plays an important role in the development of [...] Read more.

Hypoxic-ischemic brain damage (HIBD) is a common cause of death or mental retardation in newborns. Ferroptosis is a novel form of iron-dependent cell death driven by lipid peroxidation, and recent studies have confirmed that ferroptosis plays an important role in the development of HIBD. However, HIBD ferroptosis-related biomarkers remain to be discovered. An artificial neural network (ANN) was established base on differentially expressed genes (DEGs) related to HIBD and ferroptosis and validated by external dataset. The protein–protein interaction (PPI) network, support vector machine-recursive feature elimination (SVM-RFE) algorithms, and random forest (RF) algorithm were utilized to identify core genes of HIBD. An in vitro model of glutamate-stimulated HT22 cell HIBD was constructed, and glutamate-induced ferroptosis and mitochondrial structure and function in HT22 cells were examined by propidium iodide (PI) staining, flow cytometry, Fe²⁺ assay, Western blot, JC-1 kit, and transmission electron microscopy (TEM). In addition, Western blot and immunofluorescence assays were used to detect the NF-κB/STAT3 pathway. An HIBD classification model was constructed and presented excellent performance. The PPI network and two machine learning algorithms indicated two hub genes in HIBD. Lipocalin 2 (LCN2) was the core gene correlated with the risk of HIBD according to the results of differential expression analysis and logistic regression diagnostics. Subsequently, we verified in an in vitro model that LCN2 is highly expressed in glutamate-induced ferroptosis in HT22 cells. More importantly, LCN2 silencing significantly inhibited glutamate-stimulated ferroptosis in HT22 cells. We also found that glutamate-stimulated HT22 cells produced mitochondrial dysfunction. Furthermore, in vitro experiments confirmed that NF-κB and STAT3 were activated and that silencing LCN2 could have the effect of inhibiting their activation. In short, our findings reveal a molecular mechanism by which LCN2 may promote ferroptosis in HIBD through activation of the NF-κB/STAT3 pathway, providing new and unique insights into LCN2 as a biomarker for HIBD and suggesting new preventive and therapeutic strategies for HIBD. Full article

(This article belongs to the Special Issue Ferroptosis and Its Potential Role in the Physiopathology of Brain Injury)

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Graphical abstract

Graphical abstract
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Differentially expressed gene analysis and gene set enrichment analysis. (A) Distribution of samples based on the whole genome expression data. (B) The volcano plot shows up-regulated and down-regulated genes. (C) Heatmap of the 60 most differentially expressed genes based on GSE23333. Light blue is the control group, red is the treatment group, and the color intensity (from red to blue) indicates the higher to lower expression. (D) Three up-regulated pathways. (E) Three down-regulated pathways. Full article ">Figure 2
Weighted co-expression network analysis. (A) Assessment of scale-free fit index (left) and mean connectivity (right) for distinct soft threshold powers. (B) Dendrogram of clustering of differentially expressed genes based on topological overlap matrix. (C) Heatmap of the correlation between modules and sample traits. (D) Two scatter plots of gene significance for HIBD vs. module membership in the blue module and yellow module. (E) Venn plot for intersected genes in WGCNA modules and ferroptosis-related genes. Full article ">Figure 3
The construction of HIBD classification model and identification of gene biomarkers for HIBD. (A) Visualization of artificial neural network results. (B,C) The ROC curves were utilized to evaluate the performance of the HIBD classification model in training and validation datasets. (D) CytoHubba detected four ferroptosis-HIBD genes. (E) Three genes were identified by the SVM-RFE algorithm with an error of 0.0229. (F) A minimum error regression tree was established for six ferroptosis-HIBD genes. Full article ">Figure 4
Validation of the diagnostic value of the hub gene. (A) Venn diagram indicates the overlapping genes in SVM-RFE algorithm, random forest algorithm and CytoHubba. (B) The comparison of two potential biomarkers via GSE23333. (C) ROC curves of LCN2 in dataset GSE184997. (D) The expression of LCN2 in the dataset GSE184997. (E) Lollipop plot showing the correlation of immune cell infiltration and LCN2. Full article ">Figure 5
Glutamate stimulation of HT22 cells induces high expression of LCN2 in ferroptosis. Glutamate stimulation of HT22 cells with or without pre-treatment with Fer-1. (A) Representative results of annexin V-FITC/PI staining and quantitative analysis after the treatment (glutamate 10 mM) for 24 h. The mean ± SD is shown. Unpaired two-tailed t test was used for statistical analysis. *** p < 0.001. (B) Detection of C11-BODIPY 581/591 fluorescence of HT22 cells by flow cytometry. Glutamate: 10 mM, Fer-1: 10 μM, treatment time: 24 h. **: It represents the glutamate stimulation group compared with the control group or glutamate combined with Fer-1 group, p < 0.01. (C) Detection of Ferro Orange fluorescence of HT22 by multifunctional microplate detection system. Glutamate: 10 mM, Fer-1: 5 μM, treatment time: 24 h. Scale bar is 100 μm. (D) Representative TEM images of the untreated group, the glutamate-treated group and the glutamate-co-administered Fer-1 group. The red arrows represent mitochondria. Glutamate: 10 mM, Fer-1: 10 μM, treatment time: 24 h. Scale bar = 500 nm, 1 μm, 5 μm. (E) Western blotting analysis and quantitative analysis of ACSL4, 4HNE, SLC7A11, LCN2, GPX4 and GAPDH expression in HT22. Glutamate: 10 mM, Fer-1: 5 μM, treatment time: 24 h. **/***/****: It indicates that the glutamate-treated group compared to the blank control group, ** p < 0.01, *** p < 0.001, **** p < 0.0001. #/##/####: It indicates that pretreatment of the glutamate group with Fer-1 compared to that without Fer-1, # p < 0.05, ## p < 0.01, #### p < 0.0001. Two-way ANOVA multiple comparisons were performed between groups. Full article ">Figure 6
Glutamate stimulation of HT22 cells causes mitochondrial damage. For all experiments, glutamate: 10 mM, Fer-1: 5 μM, treatment time: 24 h. (A) HT22 cells were treated with indicated drugs for 24 h, and the level of mtROS was detected by flow cytometry with mtROSTM 580 (n = 3). ### p < 0.001 compared with the normal group; glutamate combined with Fer-1 group compared with glutamate group, * p < 0.05. (B) JC-1 kit was used to evaluate the effect of glutamate stimulation on MMP in HT22 cells. Red fluorescence: aggregates; green fluorescence: monomer; yellow fluorescence: combined. Scale bar is 100 μm. (C) The expression levels of Mitofusin2, VDAC and TOM20 in different groups were detected by Western blotting, and to quantify and analyze. */**: It indicates that the glutamate-treated group compared to the blank control group, * p < 0.05, ** p < 0.01. ###/####: It indicates that pretreatment of the glutamate group with Fer-1 compared to that without Fer-1, ### p < 0.001, #### p < 0.0001. Two-way ANOVA multiple comparisons were performed between groups. Full article ">Figure 7
LCN2 silencing inhibits glutamate-induced ferroptosis in HT22 cells. Using specific siRNA knockdown of LCN2, process accordingly according to grouping requirements. Glutamate: 10 mM. (A) Knockdown of LCN2 by specific si-RNAs in HT22, after 72 h, and the expression of LCN2 and GAPDH were detected by Western blotting. (B) After specific knockdown of LCN2 for 36 h, HT22 cells were treated with glutamate for 24 h, followed by flow cytometry to detect lipid peroxidation. *: p < 0.05. **: p < 0.01. (C) Representative images showing the amount of Ferro Orange in the indicated cells. Scale bar is 100 μm. (D) Levels of ACSL4, SLC7A11, FTH1, GPX4, LCN2 and GAPDH proteins in the indicated cells were assessed by Western blotting. */****: It indicates that the glutamate-treated group compared to the blank control group, * p < 0.05, **** p < 0.0001. ##/####: It indicates that pretreatment of the glutamate group with Fer-1 compared to that without Fer-1, ## p < 0.01, #### p < 0.0001. Two-way ANOVA multiple comparisons were performed between groups. Full article ">Figure 8
LCN2 regulates the activation of the NF-κB /STAT3 axis in glutamate-stimulated HT22 cells. (A–D,F–I) Western blotting was used to detect the protein expression of P-P65, P65, STAT3, P-STAT3, LCN2 and GAPDH in different treated HT22 cells. Quantitative analysis of results. Compared with the addition of the si-NC empty carrier group, * p < 0.05, *** p < 0.001, **** p < 0.0001. Compared to the group treated with glutamate stimulation combined with si-NC, # p < 0.05, ## p < 0.01, #### p < 0.0001. Two-way ANOVA multiple comparisons were performed between groups. (E) Immunofluorescence images of HT22 cells labeled with P65 antibody. Co-staining with DAPI (blue) to show the nucleus. Scale bar = 100 μm. For all experiments, glutamate: 10 mM, Fer-1: 5 μM. Full article ">

22 pages, 1498 KiB

Open AccessArticle

Phenolic Compounds Profiling and Their Antioxidant Capacity in the Peel, Pulp, and Seed of Australian Grown Avocado

by Xiaoyan Lyu, Osman Tuncay Agar, Colin J. Barrow, Frank R. Dunshea and Hafiz A. R. Suleria

Antioxidants 2023, 12(1), 185; https://doi.org/10.3390/antiox12010185 - 12 Jan 2023

Cited by 22 | Viewed by 6146

Abstract

Avocados (Persea americana M.) are highly valued fruits consumed worldwide, and there are numerous commercially available varieties on the market. However, the high demand for fruit also results in increased food waste. Thus, this study was conducted for comprehensive profiling of polyphenols [...] Read more.

Avocados (Persea americana M.) are highly valued fruits consumed worldwide, and there are numerous commercially available varieties on the market. However, the high demand for fruit also results in increased food waste. Thus, this study was conducted for comprehensive profiling of polyphenols of Hass, Reed, and Wurtz avocados obtained from the Australian local market. Ripe Hass peel recorded the highest TPC (77.85 mg GAE/g), TTC (148.98 mg CE/g), DPPH (71.03 mg AAE/g), FRAP (3.05 mg AAE/g), RPA (24.45 mg AAE/g), and ABTS (75.77 mg AAE/g) values; unripe Hass peel recorded the highest TFC (3.44 mg QE/g); and Wurtz peel recorded the highest TAC (35.02 mg AAE/g). Correlation analysis revealed that TPC and TTC were significantly correlated with the antioxidant capacity of the extracts. A total of 348 polyphenols were screened in the peel. A total of 134 compounds including 36 phenolic acids, 70 flavonoids, 11 lignans, 2 stilbenes, and another 15 polyphenols, were characterised through LC-ESI-QTOF-MS/MS, where the majority were from peels and seeds of samples extract. Overall, the hierarchical heat map revealed that there were a significant amount of polyphenols in peels and seeds. Epicatechin, kaempferol, and protocatechuic acid showed higher concentrations in Reed pulp. Wurtz peel contains a higher concentration of hydroxybenzoic acid. Our results showed that avocado wastes have a considerable amount of polyphenols, exhibiting antioxidant activities. Each sample has its unique value proposition based on its phenolic profile. This study may increase confidence in utilising by-products and encourage further investigation into avocado by-products as nutraceuticals. Full article

(This article belongs to the Special Issue Natural Products: Biological-, Antioxidant Properties and Health Effects - 2nd Edition)

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21 pages, 2544 KiB

Open AccessFeature PaperArticle

Above the Invasive and Ornamental Attributes of the Traveler’s Palm: An In Vitro and In Silico Insight into the Anti-Oxidant, Anti-Enzymatic, Cytotoxic and Phytochemical Characterization of Ravenala madagascariensis

by Shanoo Suroowan, Eulogio Jose Llorent-Martínez, Gokhan Zengin, Stefano Dall’Acqua, Stefania Sut, Kalaivani Buskaran, Sharida Fakurazi, Bao Le Van, Mohnad Abdalla, Ashraf N. Abdalla, Asaad Khalid and Mohamad Fawzi Mahomoodally

Antioxidants 2023, 12(1), 184; https://doi.org/10.3390/antiox12010184 - 12 Jan 2023

Cited by 4 | Viewed by 2953

Abstract

Ravenala madagascariensis is a widely known ornamental and medicinal plant, but with a dearth of scientific investigations regarding its phytochemical and pharmacological properties. Hence, these properties were appraised in this study. The DPPH (154.08 ± 2.43 mgTE/g), FRAP (249.40 ± 3.01 mgTE/g), CUPRAC [...] Read more.

Ravenala madagascariensis is a widely known ornamental and medicinal plant, but with a dearth of scientific investigations regarding its phytochemical and pharmacological properties. Hence, these properties were appraised in this study. The DPPH (154.08 ± 2.43 mgTE/g), FRAP (249.40 ± 3.01 mgTE/g), CUPRAC (384.57 ± 1.99 mgTE/g), metal chelating (29.68 ± 0.74 mgEDTAE/g) and phosphomolybdenum assay (2.38 ± 0.07 mmolTE/g) results demonstrated that the aqueous extract had the most prominent antioxidant activity, while the methanolic extract displayed the best antioxidant potential in the ABTS assay (438.46 ± 1.69 mgTE/g). The HPLC-ESI-Q-TOF-MS-MS analysis allowed the characterization of 41 metabolites. The methanolic extract was the most active against acetylcholinesterase. All extracts were active against the alpha-amylase and alpha-glucosidase enzymes, with the ethyl acetate extract being the most active against the alpha-amylase enzyme, while the methanolic extract showed the best alpha-glucosidase inhibition. A plethora of metabolites bonded more energetically with the assayed enzymes active sites based on the results of the in silico studies. R. madagascariensis extracts used in this study exhibited cytotoxicity against HT29 cells. The IC₅₀ of the methanolic extract was lower (506.99 ug/mL). Based on the heat map, whereby flavonoids were found to be in greater proportion in the extracts, it can be concluded that the flavonoid portion of the extracts contributed to the most activity. Full article

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28 pages, 11121 KiB

Open AccessArticle

Study of Ferroptosis Transmission by Small Extracellular Vesicles in Epithelial Ovarian Cancer Cells

by Carmen Alarcón-Veleiro, Rocío Mato-Basalo, Sergio Lucio-Gallego, Andrea Vidal-Pampín, María Quindós-Varela, Thamer Al-Qatarneh, Germán Berrecoso, Ángel Vizoso-Vázquez, María C. Arufe and Juan Fafián-Labora

Antioxidants 2023, 12(1), 183; https://doi.org/10.3390/antiox12010183 - 12 Jan 2023

Cited by 8 | Viewed by 3220

Abstract

Epithelial ovarian cancer (EOC) is the most lethal gynecological cancer. The current treatment for EOC involves surgical debulking of the tumors followed by a combination of chemotherapy. While most patients achieve complete remission, many EOCs will recur and develop chemo-resistance. The cancer cells [...] Read more.

Epithelial ovarian cancer (EOC) is the most lethal gynecological cancer. The current treatment for EOC involves surgical debulking of the tumors followed by a combination of chemotherapy. While most patients achieve complete remission, many EOCs will recur and develop chemo-resistance. The cancer cells can adapt to several stress stimuli, becoming resistant. Because of this, new ways to fight resistant cells during the disease are being studied. However, the clinical outcomes remain unsatisfactory. Recently, ferroptosis, a novel form of regulated cell death trigged by the accumulation of iron and toxic species of lipid metabolism in cells, has emerged as a promising anti-tumor strategy for EOC treatment. This process has a high potential to become a complementary treatment to the current anti-tumor strategies to eliminate resistant cells and to avoid relapse. Cancer cells, like other cells in the body, release small extracellular vesicles (sEV) that allow the transport of substances from the cells themselves to communicate with their environment. To achieve this, we analyzed the capacity of epithelial ovarian cancer cells (OVCA), treated with ferroptosis inducers, to generate sEV, assessing their size and number, and study the transmission of ferroptosis by sEV. Our results reveal that OVCA cells treated with ferroptotic inducers can modify intercellular communication by sEV, inducing cell death in recipient cells. Furthermore, these receptor cells are able to generate a greater amount of sEV, contributing to a much higher ferroptosis paracrine transmission. Thus, we discovered the importance of the sEV in the communication between cells in OVCA, focusing on the ferroptosis process. These findings could be the beginning form to study the molecular mechanism ferroptosis transmission through sEV. Full article

(This article belongs to the Special Issue Oxidative Stress and Inflammation in Cancer)

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18 pages, 1909 KiB

Open AccessArticle

Impact of Phenolic Acid Derivatives on the Oxidative Stability of β-Lactoglobulin-Stabilized Emulsions

by Alina Bock, Helena Kieserling, Ulrike Steinhäuser and Sascha Rohn

Antioxidants 2023, 12(1), 182; https://doi.org/10.3390/antiox12010182 - 12 Jan 2023

Cited by 3 | Viewed by 1955

Abstract

Proteins, such as β-lactoglobulin (β-Lg), are often used to stabilize oil–water-emulsions. By using an additional implementation of phenolic compounds (PC) that might interact with the proteins, the oxidative stability can be further improved. Whether PC have a certain pro-oxidant effect on oxidation processes, [...] Read more.

Proteins, such as β-lactoglobulin (β-Lg), are often used to stabilize oil–water-emulsions. By using an additional implementation of phenolic compounds (PC) that might interact with the proteins, the oxidative stability can be further improved. Whether PC have a certain pro-oxidant effect on oxidation processes, while interacting non-covalently (pH-6) or covalently (pH.9) with the interfacial protein-film, is not known. This study aimed to characterize the impact of phenolic acid derivatives (PCDs) on the antioxidant efficacy of the interfacial β-Lg-film, depending on their structural properties and pH-value. Electron paramagnetic resonance (EPR) analyses were performed to assess the radical scavenging in the aqueous and oil phases of the emulsion, and the complexation of transition metals: these are well known to act as pro-oxidants. Finally, in a model linseed oil emulsion, lipid oxidation products were analyzed over storage time in order to characterize the antioxidant efficacy of the interfacial protein-film. The results showed that, at pH.6, PCDs can scavenge hydrophilic radicals and partially scavenge hydrophobic radicals, as well as reduce transition metals. As expected, transition metals are complexed to only a slight degree, leading to an increased lipid oxidation through non-complexed reduced transition metals. At pH.9, there is a strong complexation between PCDs and the transition metals and, therefore, a decreased ability to reduce the transition metals; these do not promote lipid oxidation in the emulsion anymore. Full article

(This article belongs to the Special Issue Antioxidants and Oxidative Stability in Fats and Oils)

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Structures of the added phenolic acid derivatives. Full article ">Figure 2
Tempol degradation kinetic from the aqueous phase (a) of the emulsion at pH 6 and (b) at pH 9; here, the β-Lg emulsions are compared without (β-lg) and with PCD (+ PCD) addition. The starting point at t = 0 min represents the output signal of the pure radical. From t = 1 min onwards, the radical degradation or the radical regeneration is shown by an increase or decrease in the original radical signal. The values shown are an individual exemplary series of measurements. Full article ">Figure 3
16-DOXYL-stearic acid degradation kinetic from the lipid phase (a) of the emulsion at pH 6 and (b) at pH 9; Here, the β-Lg emulsions are compared without (β-lg) and with PCD (+PCD) addition. The starting point at t = 0 min represents the output signal of the pure radical. From t = 1 min onwards, the radical degradation or radical regeneration is shown by an increase or decrease in the original radical signal. Full article ">Figure 4
The EPR spectra show the change in the Cu(II) signal in the presence of the β-Lg interfacial film of an emulsion without PCD addition (top) and with the addition of CA (middle) and RA (bottom) at (a) pH 6 and (b) pH 9. The spectra shown are an individual exemplary series of measurements. Full article ">Figure 5
Fe(III)-reducing power calculated as ascorbic acid equivalents (a) of β-Lg-PCD mixtures at pH 6 and (b) at pH 9; the letters (a–g) are describing the statistical homogeneous groups without significant differences (p > 0.05) for (a,b). Full article ">Figure 6
Extinction of the emulsion oil extract at 233 nm as an indicator for the formation of conjugated dienes within the emulsion (a) at pH 6 and (b) at pH 9; peroxide values for emulsions stabilized by different PCD and stored for 21 days (c) at pH 6 and (d) at pH 9. Full article ">Figure 7
Oxidation products of the linseed oil emulsion after 21 d of storage time, shown as quotient of the internal standard: Quantitative main components of the secondary oxidation products of linolenic acid (a) at pH 6 and (b) at pH 9; the letters (a–c and A–B) are describing the statistical homogeneous groups without significant differences (p >0.05) for (a,b); and the further reaction of a secondary oxidation product ((E)-2-hexenal) to a tertiary oxidation product (2-ethylfuran) (c) at pH 6 and (d) at pH 9; the letters (a–d and A-E) are describing the statistical homogeneous groups without significant differences (p >0.05) for (c,d). Full article ">

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Antioxidants, Volume 12, Issue 1 (January 2023) – 211 articles

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