H2O2-induced Lipid Peroxidation Research Articles

Antihemolytic activity of flavonoids from butanolic extract of Carica papaya L. cultivar ‘Red Lady’ leaf

Flavonoids are well-known phytochemicals with antioxidant properties. They are an essential component of various medicinal, pharmaceutical, nutraceutical, and cosmetic products. In the current study we evaluated the erythrocyte membrane protection efficacy of flavonoid rich butanolic extract of Carica papaya L. (FBC) cultivar Red Lady young leaf against H2O2 induced hemolysis. The flavonoid fraction of C. papaya is a combination of eleven flavonoids viz., quercetin, quercetin dimer, kaempferol, luteolin and 2-(3,4-Dihydroxy-5-methoxyphenyl)- 3,5,7- trihydroxy-6-metoxy-2,3 dihydro-4H-chromen-4-one, rutin, isoquercetin, manghaslin, nictoflorin, astragalin and mauritianin. The result of the study showed that FBC significantly inhibited H2O2 induced hemolysis and lipid peroxidation in a concentration dependent manner (0.2 mg/mL, 0.4 mg/mL and 0.8 mg/mL). FBC at concentrations 0.2 mg/mL, 0.4 mg/mL and 0.8 mg/mL inhibited the hemolysis to 76 %, 85 % and 87 % respectively. Lipid peroxidation caused by H2O2 was reduced to 35 %, 45 % and 65 % with respect to increasing concentration of FBC. Osmotic fragility test of erythrocytes in hypotonic solutions showed that as compared to negative and positive controls, FBC pretreated (0.4 mg/mL) erythrocytes shifted the OFT curve towards left indicates the protection of erythrocytes from osmotic fragility. The membrane protection role of flavonoid extracts in erythrocytes were further confirmed by scanning electron microscopy (SEM), which was marked by the reduction in erythrocyte membrane blebbing and echinocyte formation

Rainbow Trout (Oncorhynchus mykiss) as Source of Multifunctional Peptides with Antioxidant, ACE and DPP-IV Inhibitory Activities.

The present study aimed at characterizing the possible biological activities of the multifunctional low molecular weight fractions (<3 kDa) peptides isolated from rainbow trout (Oncorhynchus mykiss) obtained by enzymatic hydrolysis. The fish protein hydrolysate (FPH) was tested for its antioxidant property along with its angiotensin converting enzyme (ACE) and dipeptidyl peptidase IV (DPP-IV) inhibitory activities. In particular, the 2,2-diphenyl-1-picrylhydrazyl (DPPH), the ferric reducing antioxidant power (FRAP), the oxygen radical absorbance capacity (ORAC) assay and the 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays were carried out for the evaluation of the in vitro antioxidant activity. The cell-free ACE and DPP-IV inhibitory activity assays were also estimated, showing a dose-dependent inhibition. These biological properties were additionally quantified at the cellular level using human intestinal Caco-2 cells. Namely, the antioxidant activity was determined by evaluating the capability of the hydrolysate to reduce the H2O2-induced reactive oxygen species (ROS) and lipid peroxidation levels, and the DPP-IV activity assays show a reduction of enzyme activity of up to 27.57 ± 3.7% at 5 mg/mL. The results indicate that Oncorhynchus mykiss-derived peptides may have potential employment as health-promoting ingredients.

Valorization of the Antioxidant Effect of Mantua PGI Pear By-Product Extracts: Preparation, Analysis and Biological Investigation.

For improving the management of the production chain of PGI Mantua pears (which comprises many varieties, including Abate Fetel), applying the cardinal principles of circular economy and sustainability, the fruits with diseases or defects were recovered for producing dried rounds of pears from the Abate Fetel cultivar, a new product with high nutritional value that extends the remaining life. This process led to the production of secondary and residual by-products, which are mainly composed of the highest and lowest part of the fruits, comprising seeds, pulps, peels and petioles. Hence, this study was focused on the valorization of these secondary by-products of Abate Fetel pears through the production of pear extracts using traditional and "green" extraction methods that involve the use of supercritical CO2 fluid extraction. The produced extracts, together with a reference solvent-derived extract, were analyzed by HPLC-ESI-MS, and in parallel, their direct and cellular antioxidant activity were assessed. Evidence has indicated that all the tested extracts reduced the H2O2-induced reactive oxygen species (ROS), lipid peroxidation and nitric oxide (NO) levels, respectively, in human intestinal Caco-2 cells. Hence, this study clearly suggests that extracts obtained from Mantuan PGI pear by-products may be used as valuable sources of bioactive upcycled phytocomplex for the development of dietary supplements and/or functional foods.

Potential anti-inflammatory activity of the Anastatica hierochuntica essential oil

ABSTRACT The aims of this study were to evaluate the anti-inflammatory activity and the possible anti-inflammatory mechanisms of Anastatica hierochuntica essential oil (EOAH). The essential oils were obtained by hydrodistillation of the aerial parts of the plant, and analyzed using gas chromatography and mass spectrometry (GC/MS). Their anti-inflammatory activity was evaluated using classical model of inflammation (carrageenan-induced paw edema and peritonitis). Nitric oxide radical-scavenging activity and lipid peroxidation were determined to assess the antioxidant activity of the essential oil. The EOAH mainly comprised Cuminaldehyde (33.8%), trans-β-Caryophyllene (32.5%), and Linalool (8.2%). EOAH significantly reduced edema, peritonitis, myeloperoxidase activity, and NOx-peritoneal lavage concentration induced by carrageenan. In addition, nitric oxide radical generated from sodium nitroprusside was significantly inhibited by the essential oil. Furthermore, EOAH was able to prevent Fe+2- or Fe+2 plus H2O2-induced lipid peroxidation. In conclusion, the EOAH exerts anti-inflammatory activity that could be related to its antioxidant capacity.

Exploitation of Olive (Olea europaea L.) Seed Proteins as Upgraded Source of Bioactive Peptides with Multifunctional Properties: Focus on Antioxidant and Dipeptidyl-Dipeptidase-IV Inhibitory Activities, and Glucagon-like Peptide 1 Improved Modulation.

Agri-food industry wastes and by-products include highly valuable components that can upgraded, providing low-cost bioactives or used as an alternative protein source. In this context, by-products from olive production and olive oil extraction process, i.e., seeds, can be fostered. In particular, this work was aimed at extracting and characterizing proteins for Olea europaea L. seeds and at producing two protein hydrolysates using alcalase and papain, respectively. Peptidomic analysis were performed, allowing to determine both medium- and short-sized peptides and to identify their potential biological activities. Moreover, an extensive characterization of the antioxidant properties of Olea europaea L. seed hydrolysates was carried out both in vitro by 2,2-diphenyl-1-picrylhydrazyl (DPPH), by ferric reducing antioxidant power (FRAP), and by 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assays, respectively, and at cellular level by measuring the ability of these hydrolysates to significant reduce the H2O2-induced reactive oxygen species (ROS) and lipid peroxidation levels in human intestinal Caco-2 cells. The results of the both hydrolysates showed significant antioxidant properties by reducing the free radical scavenging activities up to 65.0 ± 0.1% for the sample hydrolyzed with alcalase and up to 75.7 ± 0.4% for the papain hydrolysates tested at 5 mg/mL, respectively. Moreover, similar values were obtained by the ABTS assays, whereas the FRAP increased up to 13,025.0 ± 241.5% for the alcalase hydrolysates and up to 12,462.5 ± 311.9% for the papain hydrolysates, both tested at 1 mg/mL. According to the in vitro results, both papain and alcalase hydrolysates restore the cellular ROS levels up 130.4 ± 4.24% and 128.5 ± 3.60%, respectively, at 0.1 mg/mL and reduce the lipid peroxidation levels up to 109.2 ± 7.95% and 73.0 ± 7.64%, respectively, at 1.0 mg/mL. In addition, results underlined that the same hydrolysates reduced the activity of dipeptidyl peptidase-IV (DPP-IV) in vitro and at cellular levels up to 42.9 ± 6.5% and 38.7 ± 7.2% at 5.0 mg/mL for alcalase and papain hydrolysates, respectively. Interestingly, they stimulate the release and stability of glucagon-like peptide 1 (GLP-1) hormone through an increase of its levels up to 660.7 ± 21.9 pM and 613.4 ± 39.1 pM for alcalase and papain hydrolysates, respectively. Based on these results, olive seed hydrolysates may represent new ingredients with antioxidant and anti-diabetic properties for the development of nutraceuticals and functional foods for the prevention of metabolic syndrome onset.

Hempseed (Cannabis sativa) Peptides WVSPLAGRT and IGFLIIWV Exert Anti-inflammatory Activity in the LPS-Stimulated Human Hepatic Cell Line.

WVSPLAGRT (H2) and IGFLIIWV (H3) are two transepithelial transported intestinal peptides obtained from the hydrolysis of hempseed protein with pepsin, which exert antioxidant activity in HepG2 cells. Notably, both peptides reduce the H2O2-induced reactive oxygen species, lipid peroxidation, and nitric oxide (NO) production levels in HepG2 cells via the modulation of the nuclear factor erythroid 2-related factor 2 and the inducible nitric oxide synthase (iNOS) pathways, respectively. Due to the close link between inflammation and oxidative stress and with the objective of fostering the multifunctional behavior of bioactive peptides, in this study, the molecular characterization of the anti-inflammatory and immunomodulatory properties of H2 and H3 was carried out in HepG2 cells. In fact, both peptides were shown to modulate the production of pro (IFN-γ: −33.0 ± 6.7% H2, p = 0.011; −13.1 ± 2.0% H3, p = <0.0001; TNF: −17.6 ± 1.7% H2, p = 0.0004; −20.3 ± 1.7% H3, p = <0.0001; and IL-6: −15.1 ± 6.5% H3, p = 0.010)- and anti (IL-10: +9.6 ± 3.1% H2, p = 0.010; +26.0 ± 2.3% H3, p = < 0.0001)-inflammatory cytokines and NO (−9.0 ± 0.7% H2, p = <0.0001; −7.2 ± 1.8% H3, p = <0.0001) through regulation of the NF-κB and iNOS pathways, respectively, in HepG2 cells stimulated by lipopolysaccharides.

Investigation of the intestinal trans-epithelial transport and antioxidant activity of two hempseed peptides WVSPLAGRT (H2) and IGFLIIWV (H3)

A preceding paper has shown that a hempseed peptic hydrolysate displays a cholesterol-lowering activity with a statin-like mechanism of action in HepG2 cells and a potential hypoglycemic activity by the inhibition of dipeptidyl peptidase-IV in Caco-2 cells. In the framework of a research aimed at fostering the multifunctional behavior of hempseed peptides, we present here the identification and evaluation of some antioxidant peptides from the same hydrolysate. After evaluation of its diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, a trans-epithelial transport experiment was performed using differentiated Caco-2 cells that permitted the identification of five transported peptides that were synthesized and evaluated by measuring the oxygen radical absorbance capacity (ORAC), the ferric reducing antioxidant power (FRAP), and the 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid (ABTS), and diphenyl-2-picrylhydrazyl radical DPPH assays. The most active peptides, i.e. WVSPLAGRT (H2) and IGFLIIWV (H3), were then tested in cell assays. Both peptides were able to reduce the H2O2-induced reactive oxygen species (ROS), lipid peroxidation, and nitric oxide (NO) production levels in HepG2 cells, via the modulation of Nrf-2 and iNOS pathways, respectively.

Alendronate inhibits triglyceride accumulation and oxidative stress in adipocytes and the inflammatory response of macrophages which are associated with adipose tissue dysfunction.

Alendronate, a bisphosphonate used to prevent osteoporosis, stimulates osteogenesis but impairs adipogenesis. Different clinical trials suggest that the incidence of diabetes may be lower in patients treated with alendronate. Taking into account the importance of adipocytes and macrophages of adipose tissue in insulin resistance and type 2 diabetes, it is necessary to evaluate the effect of alendronate in both cell types. In this paper, we investigated the effect of alendronate on the differentiation to adipocytes of 3T3-L1 fibroblasts, the cell line most used to study adipogenesis, and also its effect on lipid content and oxidative stress in mature adipocytes as well as on the inflammatory response of macrophages. We found that alendronate inhibits differentiation of 3T3-L1 fibroblasts to adipocytes in keeping with reports in other cell lines. On the other hand, treatment of 3T3-L1 adipocytes with alendronate was able to decrease triglyceride content and to prevent H2O2-induced lipid peroxidation which was evaluated as an indicator of oxidative stress. In addition, it was found that activation of RAW 264.7 macrophages to a pro-inflammatory M1 type is inhibited by this bisphosphonate. These results suggest that alendronate may contribute to prevent adipocyte excessive enlargement and the induction of oxidative stress in 3T3-L1 adipocytes as well as the activation of macrophages to a pro-inflammatory M1 type, which are events associated with adipose tissue dysfunction and insulin resistance. In this study, we unraveled the underlying mechanisms of events that were previously observed in clinical trials.

A novel Keap1 inhibitor iKeap1 activates Nrf2 signaling and ameliorates hydrogen peroxide-induced oxidative injury and apoptosis in osteoblasts

An ultra-large structure-based virtual screening has discovered iKeap1 as a direct Keap1 inhibitor that can efficiently activate Nrf2 signaling. We here tested its potential effect against hydrogen peroxide (H2O2)-induced oxidative injury in osteoblasts. In primary murine and human osteoblasts, iKeap1 robustly activated Nrf2 signaling at micromole concentrations. iKeap1 disrupted Keap1-Nrf2 association, causing Nrf2 protein stabilization, cytosol accumulation and nuclear translocation in murine and human osteoblasts. The anti-oxidant response elements (ARE) activity and transcription of Nrf2-ARE-dependent genes (including HO1, NQO1 and GCLC) were increased as well. Significantly, iKeap1 pretreatment largely ameliorated H2O2-induced reactive oxygen species production, lipid peroxidation and DNA damage as well as cell apoptosis and programmed necrosis in osteoblasts. Moreover, dexamethasone- and nicotine-induced oxidative injury and apoptosis were alleviated by iKeap1. Importantly, Nrf2 shRNA or CRISPR/Cas9-induced Nrf2 knockout completely abolished iKeap1-induced osteoblast cytoprotection against H2O2. Conversely, CRISPR/Cas9-induced Keap1 knockout induced Nrf2 cascade activation and mimicked iKeap1-induced cytoprotective actions in murine osteoblasts. iKeap1 was ineffective against H2O2 in the Keap1-knockout murine osteoblasts. Collectively, iKeap1 activated Nrf2 signaling cascade to inhibit H2O2-induced oxidative injury and death of osteoblasts.

Pinus thunbergii Parl. Extracts Reduce Acute Inflammation by Targeting Oxidative Stress

Pinus thunbergii Parl. (PTP) has traditionally been used for edible and medicinal purposes to treat several disorders, including diabetes and neuralgia. Therefore, this study sought to evaluate the inhibitory effects of PTP leaf ethanol extracts on acute inflammation. Moreover, the reactive oxygen species (ROS) scavenging activity, superoxide dismutase (SOD) activity, lipopolysaccharide (LPS)-induced nitric oxide (NO) generation, and H2O2-induced lipid peroxidation capacity of PTP were assessed in vitro in RAW 264.7 macrophages. Our results suggest that PTP prevents cell damage caused by oxidative free radicals and downregulates the expression of LPS-induced inflammation-associated factors including inducible nitric oxidase synthetase (iNOS), cyclooxygenase-2 (COX-2), and prostaglandin E2 (PGE2). PTP inhibited NO production by 53.5% (P < 0.05) and iNOS expression by 71.5% (P < 0.01) at 100 µg/mL. PTP at 100 µg/mL also inhibited ROS generation by 58.2% (P < 0.01) and SOD activity by 29.3%, as well as COX-2 expression by 83.3% (P < 0.01) and PGE2 expression by 98.6% (P < 0.01). The anti-inflammatory effects of PTP were confirmed in vivo using an arachidonic acid (AA)-induced ear edema mouse model. Ear thickness and myeloperoxidase (MPO) activity were evaluated as indicators of inflammation. PTP inhibited edema formation by 64.5% (P < 0.05) at 1.0 mg/ear. A total of 16 metabolites were identified in PTP extracts and categorized into subgroups, including two phenolic acids (mainly quinic acid), seven flavonoids, five lignans, one sesquiterpenoid, and one long-chain fatty acid. Therefore, our results suggest that PTP possesses anti-inflammatory properties.

Preservation of hydrogen peroxide-induced oxidative damage in HepG-2 cells by rice protein hydrolysates pretreated with electron beams

In this paper, electron beam irradiated rice protein hydrolysates (ERPHs) were assessed for their ability to prevent hydrogen peroxide-induced oxidative stress in human HepG-2 cells. The related mechanism was also studied by analyzing the structural changes. Cytotoxicity experiments showed that rice protein hydrolysates pretreated with electron beam irradiation (EBI) were not toxic to cells if appropriate concentrations were applied. Cell viability markedly increased when the cells were treated with ERPHs before H2O2 induction. Furthermore, the ERPHs effectively suppressed H2O2-induced ROS production and lipid peroxidation and increased the protein expression levels of the intracellular antioxidant enzymes SOD, GSH-Px and CAT in H2O2-stressed HepG-2 cells. Consequently, the loss of mitochondrial membrane potential and cell apoptosis was alleviated. Circular dichroism analysis showed that pretreatment of rice protein with EBI significantly changed the secondary structure (the conversion of α-helices to random coils), which is beneficial to the improvement of its antioxidative activity. ERPHs exhibited stronger antioxidative effects than those without irradiation, possibly because of the difference in molecular weight distribution and amino acid composition. These findings indicate an efficient way to produce peptides with better antioxidant activity.

Isolation of an antioxidant peptide from krill protein hydrolysates as a novel agent with potential hepatoprotective effects

Abstract Krill accounts for the highest abundant animal biomass on earth though it remains underexplored. During the present study, krill protein hydrolysates (KPH) were prepared using several proteases, optimizing reaction time for dose-response analysis of antioxidant activity. Bioassay-guided fractionation of selected KPH of pepsin, first by ultrafiltration ( 3 kDa), revealed the active fraction, 1–3 kDa, which was consecutively separated by ion-exchange chromatography and stepwise RP-HPLC. Three peptides were identified, sequenced and synthesized. The peptide P2 was effective in reducing H2O2-induced oxidative stress and lipid peroxidation by increasing antioxidant enzyme activities, including superoxide dismutase, catalase, and glutathione peroxidase. P2 reduced apoptosis in H2O2-stimulated hepatocytes by regulating the expression levels of Bcl-2/Bax and caspase-3. P2 pre-treatment increased nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) expression in cultured hepatocytes. Furthermore, P2 induced the activation of Nrf2/HO-1 by activating the ERK pathway.

Evaluation of the protective effects of quercetin and gallic acid against oxidative toxicity in rat’s kidney and HEK-293 cells

Quercetin and gallic acid are phytochemicals with interesting pharmacological properties. We herein investigated the protective effect of quercetin (QUE) in comparison with gallic acid (GAL) against exogenously-induced oxidative damage in rats’ kidney and human embryonic kidney (HEK-293) cell lines. Adult Wistar rats were treated with QUE and GAL (50 mg/kg) separately or in combination with di-n-butylphthalate (DnBP) for 14 days; and HEK-293 cells were treated with different concentrations of GAL (25−294 μM) or QUE (2−17 μM or 28−165.43 μM) singly or in combination with H2O2 (200 μM). After treatment, the kidney and cell extracts were processed for biochemical analysis and histopathology. We found that GAL but not QUE prevented DnBP-induced increase in lipid peroxidation (2.603 ± 0.25 vs. 3.65 ± 0.21 μmol/mL). Treatment with QUE but not GAL was associated with increased plasma creatinine (729.09 ± 55.68 vs. 344.25 ± 50.78 μmol/l) and tissue malondialdehyde (3.72 ± 0.62 vs. 1.67 ± 0.47 μmol/mL) concentrations, along with histo-pathological changes such as glomerular and tubular degenerations. However, QUE exhibited wider therapeutic concentration ranges than GAL at which it inhibits lipid peroxidation in HEK-293 cells, and was found to inhibit H2O2-induced lipid peroxidation even at the lowest concentration (2 μM) that was tested (0.607 ± 0.074 vs. 0.927 ± 0.106 μmol/l). These suggest that the in vivo dosages required for the antioxidant protective effects of QUE in renal tissues are low.

Pelargonidin ameliorates acetaminophen-induced hepatotoxicity in mice by inhibiting the ROS-induced inflammatory apoptotic response

The common analgesic acetaminophen (N-acetyl-p-aminophenol, APAP) is non-toxic to the liver at therapeutic doses. However, an overdose of APAP can lead to APAP-induced liver failure, which has emerged as a serious issue in the US and Europe. Pelargonidin is an anthocyanidin found in pomegranates, plums, and various berries. Pelargonidin has strong antioxidant effects, directly scavenging superoxide radicals and inhibiting H₂O₂-induced lipid peroxidation. Focusing on these effects, we studied the preventative effect of pelargonidin on APAP-induced hepatotoxicity and its underlying mechanisms in vivo. We observed that pelargonidin mitigates serum alanine aminotransferase and aspartate aminotransferase activity, which are strongly associated with APAP-induced hepatotoxicity. We also found that pelargonidin reduced APAP-induced hepatic necrosis by removing excessive ROS. Hepatic necrosis stimulates the release of molecular pathogens that induce inflammation, which increases cell stress and can lead to apoptosis. Therefore, pelargonidin was able to reduce levels of necrosis, inflammation, and hepatocyte apoptosis. These results indicate that the administration of pelargonidin protects against APAP-induced hepatotoxicity and that it could be a novel protective strategy against APAP-induced liver failure.

Protective Effect of 3-Bromo-4,5-Dihydroxybenzaldehyde from Polysiphonia morrowii Harvey against Hydrogen Peroxide-Induced Oxidative Stress In Vitro and In Vivo.

We investigated the protective effects of 3-bromo-4,5-dihydroxybenzaldehyde (BDB) from Polysiphonia morrowii Harvey against hydrogen peroxide (H2O2)-induced apoptosis in Vero cells. BDB exhibited scavenging activity for DPPH, hydroxyl, and alkyl radicals. BDB also inhibited H2O2-induced lipid peroxidation, cell death, and apoptosis in Vero cells by inhibiting the production of ROS. To evaluate the molecular mechanisms of apoptosis inhibition, the expression of Bax/Bcl-xL and NF-κB was assessed by western blot assay. BDB significantly suppressed the cleavage of caspase-9 and PARP and reduced Bax levels in H2O2-induced Vero cells. Besides, BDB suppressed the phosphorylation of NF-κB and the translocation of p65 in H2O2-induced cells. Furthermore, we evaluated the effect of BDB on ROS production, cell death, and lipid peroxidation in an H2O2-stimulated zebrafish embryo model. Taken together, these results indicated that ROS generation and cell death were significantly inhibited by BDB in zebrafish embryos, thereby proving that BDB exerts excellent antioxidant activity in vitro and in vivo.

Neuroprotective Effects of the Amylin Analog, Pramlintide, on Alzheimer's Disease Are Associated with Oxidative Stress Regulation Mechanisms.

Administration of the recombinant analog of the pancreatic amyloid amylin, Pramlintide, has shown therapeutic benefits in aging and Alzheimer's disease (AD) models, both on cognition and amyloid-β (Aβ) pathology. However, the neuroprotective mechanisms underlying the benefits of Pramlintide remain unclear. Given the early and critical role of oxidative stress in AD pathogenesis and the known reactive oxygen species (ROS) modulating function of amyloids, we sought to determine whether Pramlintide's neuroprotective effects involve regulation of oxidative stress mechanisms. To address this, we treated APP/PS1 transgenic mice with Pramlintide for 3 months, starting at 5.5 months prior to widespread AD pathology onset, and measured cognition (Morris Water Maze), AD pathology, and oxidative stress-related markers and enzymes in vivo. In vitro, we determined the ability of Pramlintide to modulate H2O2-induced oxidative stress levels. Our data show that Pramlintide improved cognitive function, altered amyloid-processing enzymes, reduced plaque burden in the hippocampus, and regulated endogenous antioxidant enzymes (MnSOD and GPx1) and the stress marker HO-1 in a location specific manner. In vitro, Pramlintide treatment in neuronal models reduced H2O2-induced endogenous ROS production and lipid peroxidation in a dose-dependent manner. Together, these results indicate that Pramlintide's benefits on cognitive function and pathology may involve antioxidant-like properties of this compound.

Copper and iron overload protect Escherichia coli from exogenous H2O2 by modulating membrane phospholipid composition

Development of green technologies for sustainable development requires growth of bacteria in medium containing H2O2 from 50 to 150 mM that may contain millimolar level of copper (Cu) and iron (Fe). However, the mechanism of oxidative stress involving high concentration of exogenous H2O2 in bacteria and the effect of Cu and Fe overload on the same remain controversial. In the present study, we investigated the effect of millimolar concentration of Cu and Fe on H2O2-induced oxidative stress using the enterotoxigenic Escherichia coli 12566 as a model system. Our results indicate that at millimolar concentration, Fe and Cu protect E. coli from H2O2-induced oxidative stress by (1) suppressing H2O2-induced growth inhibition, (2) reducing H2O2-induced lipid peroxidation, (3) decreasing catalase activity and (4) modulating H2O2 -induced release of cytosolic Fe content in a dose-dependent manner. A previous study by our group shows that H2O2 significantly alters the phospholipid (PL) composition in E. coli by a dose-dependent increase in cardiolipin (CL) accompanied by decrease in sum total of phosphatidyl ethanolamine (PE) and phosphatidyl glycerol (PG). In the present study, we show that Cu2+ and Fe3+ at 1 mM reduce H2O2-induced change in PL composition of E. coli leading to stabilization of its membrane components. For the first time, our results show that overloading of E. coli with Cu2+ and Fe3+ activates a protective mechanism against H2O2-induced oxidative damage by modulating its membrane PL composition. Our study reveals a pivotal regulatory role of these metal ions in the sectors of sustainable technologies such as heavy metal bioremediation, sewage treatment and bioelectrochemical reactors that require microbial growth at high concentration of H2O2.

Histopathological and Biochemical Effects of Thyme Essential Oil on H2O2 Stress in Heart Tissues

Thymus algeriensis (T. algeriensis) is traditionally used in Tunisia to treat many human diseases. The aim of the present study was to investigate whether terpenes extracted from the aerial parts of T. algeriensis are potent cardioprotective agents for hydrogen peroxide (H2O2)-induced cardiotoxicity in rats. Thirty (30) rats were divided into six groups as per the experimental design: control (n = 6); 0.1 mmol/L H2O2 (LD H2O2) (n = 6); 1 mmol/L H2O2 (HD H2O2) (n = 6); oily fraction of T. algeriensis (OFTS) (180 mg/kg b.wt) (n = 6); OFTS + 0.1 mmol/L H2O2 (n = 6); and OFTS + 1 mmol/L H2O2 (n = 6). The H2O2 demonstrated concentration-dependent cardiotoxic effects in vitro. While, exposure of rats to OFTS significantly depleted H2O2-induced protein oxidation and lipid peroxidation, it raised antioxidant defence enzymes, and protected against H2O2-induced histopathological alterations. The antioxidant potential of the thyme essence was assessed by both enzymatic and non-enzymatic antioxidants. In conclusion, OFTS may be a potential compound for the therapy of oxidative stress-induced heart disease.

Four-octyl itaconate activates Keap1-Nrf2 signaling to protect neuronal cells from hydrogen peroxide

BackgroundFour-octyl itaconate (OI), the itaconate’s cell-permeable derivative, can activate Nrf2 signaling via alkylation of Keap1 at its cysteine residues. The current study tested the potential neuroprotective function of OI in hydrogen peroxide (H2O2)-treated neuronal cells.MethodsSH-SY5Y neuronal cells and epigenetically de-repressed (by TSA treatment) primary murine neurons were treated with OI and/or H2O2. Nrf2 pathway genes were examined by Western blotting assay and real-time quantitative PCR analysis. Neuronal cell death was tested by the LDH and trypan blue staining assays. Apoptosis was tested by TUNEL and Annexin V assays.ResultsIn SH-SY5Y neuronal cells and primary murine neurons, OI activated Nrf2 signaling, causing Keap1-Nrf2 disassociation, Nrf2 protein stabilization and nuclear translocation, as well as expression of Nrf2-regulated genes (HO1, NQO1 and GCLC) and ninjurin2 (Ninj2). Functional studies showed that OI attenuated H2O2-induced reactive oxygen species (ROS) production, lipid peroxidation and DNA damage as well as neuronal cell death and apoptosis. shRNA-mediated knockdown, or CRISPR/Cas9-induced knockout of Nrf2 almost abolished OI-induced neuroprotection against H2O2. Keap1 is the primary target of OI. Keap1 knockout by CRISPR/Cas9 method mimicked and abolished OI-induced actions in SH-SY5Y cells. Introduction of a Cys151S mutant Keap1 in SH-SY5Y cells reversed OI-induced Nrf2 activation and anti-H2O2 neuroprotection.ConclusionsOI activates Keap1-Nrf2 signaling to protect SH-SY5Y cells and epigenetically de-repressed primary neurons from H2O2 in vitro.

Neuroligin-3 protects retinal cells from H2O2-induced cell death via activation of Nrf2 signaling

Intensified oxidative stress can cause severe damage to human retinal pigment epithelium (RPE) cells and retinal ganglion cells (RGCs). The potential effect of neuroligin-3 (NLGN3) against the process is studied here. Our results show that NLGN3 efficiently inhibited hydrogen peroxide (H2O2)-induced death and apoptosis in human RPE cells and RGCs. H2O2-induced reactive oxygen species (ROS) production, lipid peroxidation and DNA damage in retinal cells were alleviated by NLGN3. NLGN3 activated nuclear-factor-E2-related factor 2 (Nrf2) signaling, enabling Nrf2 protein stabilization, nuclear translocation and expression of key anti-oxidant enzymes (HO1, NOQ1 and GCLC) in RPE cells and RGCs. Further results demonstrate that NLGN3 activated Akt-mTORC1 signaling in retinal cells. Conversely, Akt-mTORC1 inhibitors (RAD001 and LY294002) reduced NLGN3-induced HO1, NOQ1 and GCLC mRNA expression. Significantly, Nrf2 silencing by targeted shRNAs reversed NLGN3-induced retinal cytoprotection against H2O2. We conclude that NLGN3 activates Nrf2 signaling to protect human retinal cells from H2O2. NLGN3 could be further tested as a valuable retinal protection agent.