Oxidative Stress In Caco-2 Cells Research Articles

Bioactive compounds of Citrus hystrix peel ethanolic extract and their antioxidant potential under hydrogen peroxide-induced oxidative stress in Caco-2 cells

Citrus hystrix is a fruit native to tropical Southeast Asia and Southern China. The fruits and leaves are well-known spice ingredients in Asian dishes, but the peels are usually discarded, producing a large amount of waste despite the presence of phenolic compounds. Phenolic compounds are essential for scavenging reactive oxygen species (ROS) generated naturally during metabolism. This study aimed to investigate the profile of phenolic compounds (total phenolic content, total flavonoid content, and individual phenolic compounds) and antioxidant activities (2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, ferric reducing antioxidant power, and oxygen radical absorbance capacity) of Citrus hystrix peel ethanolic extract (CHPPE). Their functional properties in a biological model (cell viability and intracellular ROS) were also evaluated using Caco-2 cells under H2O2-induced oxidative stress. We identified 37 compounds, including flavonoids (26), phenolic acids (3), stilbenes (2), lignans (1), and other phenolic compounds (5). CHPPE contained (mg/g CHPPE) hesperidin (16.10±0.05), diosmin (1.95±0.08), neohesperidin (23.83±0.06), and naringin (2.39±0.03). CHPPE exhibited remarkable antioxidant activity, which significantly improving cell viability and reducing the presence of intracellular ROS. The cytoprotective effect of CHPPE was higher than that of ascorbic acid, indicating a strong natural antioxidative capacity. Our research reveals the potential of CHPPE as a functional ingredient in the food and pharmaceutical industries.

The characterization of the pectin/alginate nanoparticle for encapsulation of hydroxypropyl-β-cyclodextrin-complexed naringin and its effects on cellular uptake and oxidative stress in Caco-2 cells

Naringin (NR) and hydroxypropyl-β-cyclodextrin (HPCD) can form a water-soluble complex, but it is unstable. This study aimed to investigate the characterization of the pectin/alginate hydrogel nanoparticles (HNPs) loading HPCD-complexed naringin. The encapsulation efficiency and loading capacity of the HNPs for NR were found to be 79.23 % ± 1.31 % and 23.79 % ± 0.67 %, respectively. HNPs had an average diameter of 409.5 ± 8.5 nm, a PDI of 0.237 ± 0.014, and a zeta-potential of −33.5 ± 0.2. FTIR, XRD, and DSC analysis confirmed that the NR-HPCD complex was embedded into the HNPs. In simulated gastrointestinal digestion, the HNPs exhibited a lower cumulative release rate compared to free NR. In Caco-2 cells, the HNPs were more efficiently transported into the cells. Consequently, the HNPs resulted in a greater decrease in ROS levels, more recovery of mitochondrial membrane potential and higher content of glutathione. This study provided a carrier for encapsulating NR, making it possible for use in food or functional food.

The Peptide Fractions of Cheddar Cheese Made with Lactobacillus helveticus 1.0612 Play Protective Effects in H2O2-Induced Oxidative-Damaged Caco-2 Cells Models.

In this study, water-soluble peptide (WSP) fractions of cheddar cheese made with Lactobacillus helveticus 1.0612 were purified into WSP-Ⅰ (<3 kDa), WSP-Ⅱ (3-10 kDa), and WSP-Ⅲ (>10 kDa). The protective effects of WSP, WSP-Ⅰ, WSP-Ⅱ, and WSP-Ⅲ fractions against oxidative stress in Caco-2 cells were assayed, and the cytoprotective mechanism of WSP-Ⅰ on cells oxidative damage was elucidated via metabolomics. The results showed that all four peptide fractions were able to attenuate the decrease in cell viability caused by oxidative stress and also could reduce the production of reactive oxygen species and malondialdehyde caused by oxidative stress, and increased cellular catalase and superoxide dismutase activities, thereby enhancing cellular antioxidant capacity. The WSP-Ⅰ fraction with the highest protective effect was used for metabolomics analysis, and 15 significantly different metabolites were screened. Functional pathway analysis revealed that the protective effect of the WSP-I fraction was related with nine metabolic pathways and weakened the metabolic disorders caused by H2O2 via regulating energy metabolism and amino acid metabolism. All in all, peptide fractions of cheddar cheese showed a cytoprotective effect through improved cellular metabolism.

Exosomes derived from EphB2-overexpressing bone marrow mesenchymal stem cells regulate immune balance and repair barrier function

Disruption of intestinal barrier function and an imbalance in intestinal immunity are crucial for the occurrence and development of ulcerative colitis. Because of their important roles in regulating inflammation and immunity, exosomes (Exos) released from bone marrow mesenchymal stem cells (BMSCs) may be useful for treating ulcerative colitis. The EphB/EphrinB signaling pathway plays a crucial role in the inflammatory process and the development and function of immune cells, and can mediate long-distance intercellular communication through extracellular vesicles. This study was conducted to explore the effects of pre-modified BMSC-Exos expressing EphB2 (EphB2-Exos) on immunoregulation in vitro. We transfected a lentivirus vector encoding EphB2 into BMSCs and isolated EphB2-Exos from the culture supernatant. Inflammation and oxidative damage in the human colon adenocarcinoma cell line (Caco-2) were induced by dextran sulfate sodium/hydrogen peroxide. In addition, spleen CD4+ T lymphocytes of rats were sorted in vitro. We conducted a series of experiments to explore the biological functions of EphB2-Exos. EphB2-Exos were successfully isolated and were found to significantly protect the activity, proliferation, and migration of Caco-2 cells that were inhibited by dextran sulfate sodium. EphB2-Exos alleviated inflammation and apoptosis and increased the activity of antioxidant enzymes while inhibiting oxidative stress in Caco-2 cells. EphB2-Exos restored intestinal barrier function by inhibiting the RhoA/ROCK pathway and regulated the polarization of CD4+T cells. EphB2-Exos enhanced intestinal barrier function and regulated the immune balance by inhibiting the RhoA/ROCK pathway in vitro. These findings suggest that EphB2-Exos can be applied as a cell-free therapy for ulcerative colitis.

P12-05 Antioxidant response of culture extracts CB08026-SYP and CB08100-SYP from species of genera Burkholderia and Tritonibacter to experimental oxidative stress in Caco-2 cells

P12-05 Antioxidant response of culture extracts CB08026-SYP and CB08100-SYP from species of genera Burkholderia and Tritonibacter to experimental oxidative stress in Caco-2 cells

Glucose Uptake and Oxidative Stress in Caco-2 Cells: Health Benefits from Posidonia oceanica (L.) Delile.

Posidonia oceanica (L.) Delile is an endemic Mediterranean marine plant of extreme ecological importance. Previous in vitro and in vivo studies have demonstrated the potential antidiabetic properties of P. oceanica leaf extract. Intestinal glucose transporters play a key role in glucose homeostasis and represent novel targets for the management of diabetes. In this study, the ability of a hydroalcoholic P. oceanica leaf extract (POE) to modulate intestinal glucose transporters was investigated using Caco-2 cells as a model of an intestinal barrier. The incubation of cells with POE significantly decreased glucose uptake by decreasing the GLUT2 glucose transporter levels. Moreover, POE had a positive effect on the barrier integrity by increasing the Zonulin-1 levels. A protective effect exerted by POE against oxidative stress induced by chronic exposure to high glucose concentrations or tert-butyl hydroperoxide was also demonstrated. This study highlights for the first time the effect of POE on glucose transport, intestinal barrier integrity, and its protective antioxidant effect in Caco-2 cells. These findings suggest that the P. oceanica phytocomplex may have a positive impact by preventing the intestinal cell dysfunction involved in the development of inflammation-related disease associated with oxidative stress.

Anticancer Potential of Post-Fermentation Media and Cell Extracts of Probiotic Strains: An In Vitro Study.

Simple SummaryProbiotics and their metabolites are very important for human health. The aim of this research was to determine probiotic strains with the strongest inhibitory properties against intestinal cancer cells. As a result of the screening, it was possible to find two strains, i.e., Lactiplantibacillus plantarum 0991 and Levilactobacillus brevis 0983, that could inhibit the proliferation of cancer cells by induction of oxidative stress and programmed cell death. Both strains exhibit interesting anticancer properties and potential as functional food ingredients; however, the results must be confirmed in further research.Background: Lactic acid bacteria (LAB), many of which are probiotics, can produce health-promoting metabolites (postbiotics). Purpose: To assess the mechanism of antiproliferative action of postbiotics, post-fermentation media (PFM) and cell extracts (CEs) of several strains of LAB were studied against colon (Caco-2), and cervix (HeLa) cancer cell lines, as well as normal intestine (IEC-6) cells, were used as a comparison. Methods: Postbiotics of various LAB (n = 39) were screened for their antiproliferative activity. The effect of PFM and CEs on reactive oxygen species (ROS), mitochondrial membrane potential (MMP), ATP production, phosphatidylserine (PS) externalisation, and apoptosis-related caspases 3/7 and 9 activation was assayed. Results: PFM and CEs showed strong dose-dependent antiproliferative activity against Caco-2 cells, up to 77.8 ± 0.8% and 58.4 ± 1.6% for PFM and CEs, respectively. Stronger inhibitory activity against cancerous (Caco-2 and HeLa) cells than against normal (IEC-6) cells was observed. PFM were more inhibitory than CEs, and both generated oxidative stress in Caco-2 cells. PFM of L. plantarum 0991 and L. brevis 0983 induced apoptosis in Caco-2 cells by the mitochondrial signalling pathway. Conclusions: Anticancer activity of PFM and CEs of LAB, as well as the ability of apoptosis induction, is strain-specific.

Decabromodiphenyl ether induces ROS-mediated intestinal toxicity through the Keap1-Nrf2 pathway.

Polybrominated diphenyl ethers (PBDEs) arewidely used brominated flame retardants as commercial products. PBDEs have been demonstrated to induce hepatic, reproductive, neural, and thyroid toxicity effects. This study aimed to clarify the potential intestinal toxicity effects of decabrominated diphenyl ether (PBDE-209) in vivo and in vitro. First, we investigated the change of PBDE-209 on oxidative stress in the intestine of mice. Subsequently, the potential toxicity mechanism of PBDE-209 in vitro was investigated. Caco-2 cells were treated with different concentrations of PBDE-209 (1, 5, and 25 μmol/L) for 24 and 48 h. We determined the cell viability, reactive oxygen species (ROS) level, multiple cellular parameters, and relative mRNA expressions. The results showed that PBDE-209 significantly injured the colon of mice, increased the intestinal levels of malondialdehyde (MDA), and changed the antioxidant enzyme activities. PBDE-209 inhibited the proliferation and induced cytotoxicity of Caco-2 cells. The change in ROS production and mitochondrial membrane potential (MMP) revealed that PBDE-209 caused oxidative stress in Caco-2 cells. The real-time PCR assays revealed that PBDE-209 inhibited the mRNA expression level of antioxidative defense factor, nuclear factor erythroid 2-related factor 2 (Nrf2). Furthermore, the FAS and Cytochrome P450 1A1 (CYP1A1) mRNA expression levels were increased in Caco-2 cells. These results suggested that PBDE-209 exerts intestinal toxicity effects in vivo and in vitro and inhibits the antioxidative defense gene expression in Caco-2 cells. This study provides an opportunity to advance the understanding of toxicity by the persistent environmental pollutant PBDE-209 to the intestine.

Ammonia impairs tight junction barriers by inducing mitochondrial dysfunction in Caco-2 cells.

Ammonia is one of the major metabolites produced by intestinal microorganisms; however, its role in intestinal homeostasis is poorly understood. The present study investigated the regulation of intestinal tight junction (TJ) proteins by ammonia and the underlying mechanisms in human intestinal Caco-2 cells. Ammonia (15, 30, and 60mM) increased the permeability of the cells in a dose-dependent manner, as indicated by reduced transepithelial electrical resistance and increased dextran flux. Immunoblot and immunofluorescence analyses revealed that the ammonia-induced increase in TJ permeability reduced the membrane localization of TJ proteins such as zonula occludens (ZO)1, ZO2, occludin, claudin-1, and claudin-3. DNA microarray analysis identified a biological pathway "response to reactive oxygen species" enriched by ammonia treatment, indicating the induction of oxidative stress in the cells. Ammonia treatment also increased the malondialdehyde content and decreased the ratio of reduced to oxidized glutathione. Meanwhile, ammonia treatment-induced mitochondrial dysfunction, as indicated by the downregulation of genes associated with the electron transport chain, reduction of the cellular ATP, NADH, and tricarboxylic acid cycle intermediate content, and suppression of the mitochondrial membrane potential. In contrast, N-acetyl cysteine reversed the ammonia-induced impairment of TJ permeability and structure without affecting the mitochondrial parameters. Collectively, ammonia impaired the TJ barrier by increasing oxidative stress in Caco-2 cells. A mitochondrial dysfunction is possibly an event preceding ammonia-induced oxidative stress. The findings of this study could potentially improve our understanding of the interplay between intestinal microorganisms and their hosts.

Cystine reduces tight junction permeability and intestinal inflammation induced by oxidative stress in Caco-2 cells

Intestinal oxidative stress produces pro-inflammatory cytokines, which increase tight junction (TJ) permeability, leading to intestinal and systemic inflammation. Cystine (Cys2) is a substrate of glutathione (GSH) and inhibits inflammation, however, it is unclear whether Cys2 locally improves intestinal barrier dysfunction. Thus, we investigated the local effects of Cys2 on oxidative stress-induced TJ permeability and intestinal inflammatory responses. Caco-2 cells were cultured in a Cys2-supplemented medium for 24 h and then treated with H2O2 for 2 h. We assessed TJ permeability by measuring transepithelial electrical resistance and the paracellular flux of fluorescein isothiocyanate–dextran 4 kDa. We measured the concentration of Cys2 and GSH after Cys2 pretreatment. The mRNA expression of pro-inflammatory cytokines was assessed. In addition, the levels of TJ proteins were assessed by measuring the expression of TJ proteins in the whole cells and the ratio of TJ proteins in the detergent-insoluble fractions to soluble fractions (IS/S ratio). Cys2 treatment reduced H2O2-induced TJ permeability. Cys2 did not change the expression of TJ proteins in the whole cells, however, suppressed the IS/S ratio of claudin-4. Intercellular levels of Cys2 and GSH significantly increased in cells treated with Cys2. Cys2 treatment suppressed the mRNA expression of pro-inflammatory cytokines, and the mRNA levels were significantly correlated with TJ permeability. In conclusion, Cys2 treatment locally reduced oxidative stress-induced intestinal barrier dysfunction possively due to the mitigation of claudin-4 dislocalization. Furthermore, the effect of Cys2 on the improvement of intestinal barrier function is related to the local suppression of oxidative stress-induced pro-inflammatory responses.

Exosomes Derived from EphB2-Overexpressing Bone Marrow Mesenchymal Stem Cells Regulate Immune Balance and Repair Barrier Function &lt;i&gt; in vitro&lt;/i&gt;

Background: Disruption of intestinal barrier function and an imbalance in intestinal immunity are crucial for the occurrence and development of ulcerative colitis. Because of their important roles in regulating inflammation and immunity, exosomes (Exos) released from bone marrow mesenchymal stem cells (BMSCs) may be useful for treating ulcerative colitis. The EphB/EphrinB signaling pathway plays a crucial role in the inflammatory process and the development and function of immune cells, and can mediate long-distance intercellular communication through extracellular vesicles. This study was conducted to explore the effects of pre-modified BMSC-Exos expressing EphB2 (EphB2-Exos) on immunoregulation in vitro. Method: sWe transfected a lentivirus vector encoding EphB2 into BMSCs and isolated EphB2-Exos from the culture supernatant. Inflammation and oxidative damage in the human colon adenocarcinoma cell line (Caco-2) were induced by dextran sulfate sodium/hydrogen peroxide. In addition, spleen CD4+ T lymphocytes of rats were sorted in vitro. We conducted a series of experiments to explore the biological functions of EphB2-Exos. Results: EphB2-Exos were successfully isolated and were found to significantly protect the activity, proliferation, and migration of Caco-2 cells that were inhibited by dextran sulfate sodium. EphB2-Exos alleviated inflammation and apoptosis and increased the activity of antioxidant enzymes while inhibiting oxidative stress in Caco-2 cells. EphB2-Exos restored barrier function by inhibiting the RhoA/ROCK pathway and regulated the polarization of CD4+T cells. Conclusion: EphB2-Exos enhanced intestinal barrier function and regulated the immune balance by inhibiting the RhoA/ROCK pathway in vitro. These findings suggest that EphB2-Exos can be applied as a cell-free therapy for ulcerative colitis.

The impact of boiling and in vitro human digestion of Solanum nigrum complex (Black nightshade) on phenolic compounds bioactivity and bioaccessibility

Solanum nigrum complex (Black nightshade) is a wild leafy vegetable with phenolic antioxidant compounds related to the reduction of oxidative stress. Changes in phenolics and bioactivity due to cooking and gastrointestinal digestion of black nightshade were compared to spinach. Predominant compounds of black nightshade were myricetin, quercetin-3-O-robinoside, 3,4-dicaffeoylquinic acid, 3-caffeoylquinic acid, and rutin, which were improved after boiling but reduced after in vitro digestion. Phenolics were reduced after digestion of black nightshade and spinach; however, bioactivity was still retained, especially in preventing oxidative stress in Caco-2 cells. Hence, indicating their potential to reduce oxidative stress related diseases of the digestive tract.

Biotechnological re-cycling of apple by-products: A reservoir model to produce a dietary supplement fortified with biogenic phenolic compounds

This study is an example of apple by-products (AP) recycling through a designed fermentation by selected autochthonous Lactobacillus plantarum AFI5 and Lactobacillus fabifermentans ALI6 used singly or as binary cultures with the selected Saccharomyces cerevisiae AYI7. Compared to Raw-, Unstarted- and Chemically Acidified-AP, Fermented-AP promoted the highest levels of total and insoluble dietary fibers, DPPH scavenging capacity, and free phenolics. The binary culture of L. plantarum AFI5 and S. cerevisiae AYI7 had the best effect on the bioavailability phenolic compounds as resulted by the LC-MS/MS validated method. The accumulation of phenolic acids derivatives highlighted the microbial metabolism during AP fermentation. Bio-converted phenolics were likely responsible for the increased DPPH scavenging capacity. The potential health-promoting effects of Fermented-AP were highlighted using Caco-2 cells. With variations among single and binary cultures, fermented-AP counteracted the inflammatory processes and the effects of oxidative stress in Caco-2 cells, and preserved the integrity of tight junctions.

Toxic effects and mechanisms of three commonly used fungicides on the human colon adenocarcinoma cell line Caco-2

Fungicides, usually refer to the chemical agents that can effectively control or kill the pathogenic microorganisms. Here, we revealed the effects of three different fungicides, imazalil (IMZ), chlorothalonil (CTL) and carbendazim (CBZ), which are typical broad-spectrum fungicides that are detected at high levels in the natural environment, on heterogeneous human epithelial colorectal cells (Caco-2 cells). All three fungicides had the potential to induce different degrees of toxicity, cause apoptosis, reactive oxygen species (ROS) and even change the cell cycle in the cells. The half maximal inhibitory concentration (IC50) of CTL is the lowest among these three fungicides, suggesting that it may have the highest exposure risk, followed by IMZ, and CBZ. The results of the real-time PCR, Western blotting, and mitochondrial membrane potential (MMP) assays and the activities of key enzymes suggested that CTL induced apoptosis in Caco-2 cells via a mitochondrial-dependent pathway, as indicated by the upregulation of the expression of the apoptotic p53 and bax genes, the increase of the apoptosis marker cytochrome-c, the decrease of mRNA level of bcl-2 gene, and the decrease in the MMP. Exposure to two other fungicides also upregulated the transcriptional level of bax and the expression of cytochrome-c, but the mRNA level of bcl-2 was increased (IMZ) or unchanged (CBZ), suggesting that other pathways may be involved in the induction of cellular apoptosis by these two fungicides. In addition, all three of the fungicides could induce oxidative stress in Caco-2 cells. Our data showed that the three different kinds of fungicides all caused toxic effects in Caco-2 cells through various pathways.

A Novel Pectic Polysaccharide of Jujube Pomace: Structural Analysis and Intracellular Antioxidant Activities.

After extraction from jujube pomace and purification by two columns (DEAE-Sepharose Fast Flow and Sepharcyl S-300), the structure of SAZMP4 was investigated by HPGPC, GC, FI-IR, GC-MS, NMR, SEM, and AFM. Analysis determined that SAZMP4 (Mw = 28.94 kDa) was a pectic polysaccharide mainly containing 1,4-linked GalA (93.48%) with side chains of 1,2,4-linked Rha and 1,3,5-linked Ara and terminals of 1-linked Rha and 1-linked Ara, which might be the homogalacturonan (HG) type with side chains of the RG-I type, corresponding to the results of NMR. In AFM and SEM images, self-assembly and aggregation of SAZMP4 were respectively observed indicating its structural features. The antioxidant activity of SAZMP4 against H2O2-induced oxidative stress in Caco-2 cells was determined by activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) as well as malondialdehyde (MDA) and reactive oxygen species (ROS) levels, indicating SAZMP4 can be a natural antioxidant. Also, a better water retention capacity and thermal stability of SAZMP4 was observed based on DSC analysis, which could be applied in food industry as an additive.

Transcriptomic responses of Caco-2 cells to Lactobacillus rhamnosus GG and Lactobacillus plantarum J26 against oxidative stress

Oxidative stress is the basic reason for aging and age-related diseases. In this study, we investigated the protective effect of 2 strains of lactic acid bacteria (LAB), Lactobacillus rhamnosus GG and L. plantarum J26, against oxidative stress in Caco-2 cells, and gave an overview of the mechanisms of lactic acid bacteria antioxidant activity using digital gene expression profiling. The 2 LAB strains provided significant protection against hydrogen peroxide (H2O2)-induced reduction in superoxide dismutase activity and increase in glutathione peroxidase activity in Caco-2 cells. However, inactive bacteria had little effect on alleviating oxidation stress in Caco-2 cells. Eight genes related to oxidative stress-FOSB, TNF, PPP1R15A, NUAK2, ATF3, TNFAIP3, EGR2, and FBN2-were significantly upregulated in H2O2-induced Caco-2 cells compared with untreated Caco-2 cells. After incubation of the H2O2-induced Caco-2 cells with L. rhamnosus GG and L. plantarum J26, 5 genes (TNF, EGR2, NUAK2, FBN2, and TNFAIP3) and 2 genes (NUAK2 and FBN2) were downregulated, respectively. In addition, the Kyoto Encyclopedia of Genes and Genomes indicated that some signaling pathways associated with inflammation, immune response, and apoptosis, such as Janus kinase/signal transducers and activators of transcription (Jak-STAT), mitogen-activated protein kinase (MAPK), nuclear factor-κB, and tumor necrosis factor, were all negatively modulated by the 2 strains, especially L. rhamnosus GG. In this paper, we reveal the mechanism of LAB in relieving oxidative stress and provide a theoretical basis for the rapid screening and evaluation of new LAB resources.

Antioxidant activity and protective effects of Saccharomyces cerevisiae peptide fractions against H2O2-induced oxidative stress in Caco-2 cells

Oxidative stress is one of the most causes of some severe diseases and the spoilage of foods. Protein hydrolysates containing antioxidant peptides are a suitable candidate to replace synthetic antioxidant. In the present study, the in vitro and cellular antioxidant properties of Saccharomyces cerevisiae protein hydrolysate and the peptide fractions have been reported. The peptide protective role was evaluated in H2O2-stimulated Caco-2 cells to consider cell viability, cellular lipid and protein oxidation, and the level of cellular antioxidant enzymes such as Catalase (CAT) and Glutathione-S-transferase (GST). The peptide fractions showed a significant ferric reducing antioxidant power (67.10–93.52 µm FeSO4/mg protein), and < 3 kDa peptide fraction with 59.5% inhibitory effect on the 7th day, exhibited the most inhibitory activity toward linoleic acid peroxidation. Ultrafiltered fractions ( < 3 kDa and 3–5 kDa) significantly (P ≤ 0.05) decreased the levels of malondialdehyde (MDA) and protein carbonyl and the production of CAT and GST enzymes as protective responses of cells under oxidative stress by H2O2. This study confirms the antioxidant activity of yeast protein hydrolysate and peptide fractions and their potential to reduce cellular oxidative stress and thus validates their potential use as a valuable ingredient of functional foods.

Reduced nitrogen supply enhances the cellular antioxidant potential of phenolic extracts through alteration of the phenolic composition in lettuce (Lactuca sativa L.).

Nitrogen availability is an important environmental factor that determines the production of phenolic compounds in vegetables, but the relationship between low nitrogen-induced alterations of phenolic compounds in vegetable crops and the cellular antioxidant activities of these compounds remains unclear. This study investigated the effect of reduced nitrogen supply (0.05 mmol L-1 nitrate) on phenolic metabolism in lettuce and the protective role of phenolic extracts against H2 O2 -induced oxidative stress in Caco-2 cells by determining cell damage, reactive oxygen species (ROS) content and antioxidant enzyme activities. Reduced nitrogen supply significantly improved the accumulation of phenolic compounds in lettuce, which was partially correlated with the upregulation of genes related to the phenolic synthesis pathway. Phenolic extracts from lettuce cultivated in low-nitrogen medium exhibited a better protective effect against H2 O2 -induced oxidative damage in Caco-2 cells than those from lettuce cultivated with adequate nitrogen. These extracts act by increasing the activities of antioxidant enzymes and, subsequently, by inhibiting ROS overproduction, which leads to a decrease in mitochondrial membrane and DNA damage. The results of HPLC and correlation analyses implied that the improvement in the protective capacity of lettuce extracts after low-nitrogen treatment may be related, not only to the increased content of phenolic compounds, but also to the increased percentage contribution of chlorogenic acid and quercetin derivatives to the total phenolic content. Reduction in nitrogen supply can be a powerful strategy to modify phenolic metabolism and composition in lettuce and, consequently, to improve their antioxidant capacity. © 2019 Society of Chemical Industry.

Raspberry extract prevents NLRP3 inflammasome activation in gut epithelial cells induced by pathogenic Escherichia coli

Abstract Raspberry extract is known for its anti-inflammatory effects, but its effectiveness against pathogenic Escherichia coli-induced inflammasome activation in epithelial cells is largely unknown. This study showed that raspberry extract treatment prevented oxidative stress in Caco-2 cells infected by E. coli O157:H7 and had prophylactic effects against E. coli O157:H7-induced NLRP3 inflammasome activation. In addition, raspberry extract mitigated reduction of nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2), a key transcription factor regulating antioxidant protein expression and nitric oxide production in Caco-2 cells upon E. coli O157:H7 infection. However, raspberry extract failed to restore NRF2 nuclear content or nitric oxide production in Caco-2 cells in the presence of aminoguanidine hydrochloride, an iNOS inhibitor. Collectively, raspberry extract treatment suppressed NLRP3 mediated inflammatory response and oxidative stress in Caco-2 cells induced by E. coli O157:H7 infection. These protective effects were partially mediated by enhanced NRF2 signaling pathway and nitric oxide production.

In vitro antioxidant activities of Rhodobacter sphaeroides and protective effect on Caco-2 cell line model.

The present study aimed to evaluate the in vitro antioxidant activities and the protective effect of Rhodobacter sphaeroides on H2O2-induced oxidative stress in Caco-2 cells. The results showed that the antioxidant action of R. sphaeroides varied with different cell concentrations and treatments. Also, the intact cells and intracellular cell-free extracts showed better antioxidant activities. Caco-2 cell-based oxidative stress model was developed by optimizing H2O2 concentration and culture time with the half lethal dose and methyl thiazolyl tetrazolium. By increasing the activity of endogenous antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase, upregulating the antioxidant ability of the anti-superoxide anion and anti-hydroxyl radical, R. sphaeroides, especially the mutant strain R. sphaeroides (CGMCC No. 8513), exhibited significant protective activity against H2O2-induced oxidative stress in Caco-2 cells. Taken together, R. sphaeroides (CGMCC No. 8513) exhibits strong antioxidant activities and is a candidate to be investigated as a potential probiotic in the future.