Protective effect of whey protein hydrolysates on H2O2-induced PC12 cells oxidative stress via a mitochondria-mediated pathway

Whey protein hydrolysates (WPHs) were prepared with pepsin and trypsin. A PC12 cell model was built to observe the protective effect of WPHs against H₂O₂-induced oxidative stress. The results indicated that WPHs reduced apoptosis by 14% and increased antioxidant enzyme activities. Flow cytometry was used to assess the accumulation of reactive oxygen species (ROS), Ca²⁺ levels and the mitochondrial membrane potential (MMP). The results showed that WPHs suppressed ROS elevation and Ca²⁺ levels and stabilised MMP by 16%. The anti-apoptosis/pro-apoptosis proteins Bcl-2/Bax and poly (ADP-ribose) polymerase (PARP) were investigated by Western-blot analysis, which indicated that WPHs increased the expression of Bcl-2 while inhibiting the expression of Bax and the degradation of PARP. WPHs also blocked Caspase-3 activation by 62%. The results demonstrate that WPHs can significantly protect PC12 cells against oxidative stress via a mitochondria-mediated pathway. These findings indicate the potential benefits of WPHs as valuable food antioxidative additives.     

Oryzadine,a new alkaloid of Oryza sativa cv. Heugjinjubyeo, attenuates oxidative stress-induced cell damage via a radical scavenging effect

The antioxidant properties of oryzadine, a new alkaloid, obtained from Oryza sativa cv. Heugjinjubyeo was investigated by applying various methods based on cell-free and cell experiments. Oryzadine showed scavenging effects on the hydroxyl radical, superoxide radical, and intracellular reactive oxygen species (ROS). Oryzadine inhibited H₂O₂-induced DNA damage, which was demonstrated by DNA tail formation, lipid peroxidation which was demonstrated by the formation of thiobarbituric acid reactive substance (TBARS), and protein oxidation which was demonstrated by protein carbonyl formation. Therefore, oryzadine protected H₂O₂-induced cell damage. Our results show that the cytoprotective effects of oryzadine stem from its ability to inhibit H₂O₂-induced apoptosis, as demonstrated by a decrease in apoptotic body formation and the inhibition of mitochondrial membrane potential (ΔΨ) loss. Taken together, these findings suggest that oryzadine protected cells against H₂O₂-induced cell damage via ROS scavenging effect. Therefore, oryzadine could be considered a significant natural source of antioxidant.     

Antioxidant Activity of Oxygen Evolving Enhancer Protein 1 Purified from Capsosiphon fulvescens

This study was conducted to determine the antioxidant activity of a protein purified from Capsosiphon fulvescens. The purification steps included sodium acetate (pH 6) extraction and diethylaminoethyl‐cellulose, reversed phase Shodex C4P‐50 column chromatography. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis indicated that the molecular weight of the purified protein was 33 kDa. The N‐terminus and partial peptide amino acid sequence of this protein was identical to the sequence of oxygen evolving enhancer (OEE) 1 protein. The antioxidant activity of the OEE 1 was determined in vitro using a scavenging test with 4 types of reactive oxygen species (ROS), including the 2,2‐diphenyl‐1‐picrylhydrazyl radical, hydroxyl radical, superoxide anion, and hydrogen peroxide (H₂O₂). OEE 1 had higher H₂O₂ scavenging activity, which proved to be the result of enzymatic antioxidants rather than nonenzymatic antioxidants. In addition, OEE 1 showed less H₂O₂‐mediated ROS formation in HepG2 cells. In conclusion, this study demonstrates that OEE 1 purified from C. fulvescens is an excellent antioxidant.     

Essential rosemary oil protects testicular cells against DNA-damaging effects of H2O2 and DMNQ

Rosemary oil (RO) is popular in the Mediterranean region as a culinary additive which has the ability to protect delicate organs such as liver, brain and heart. We examined the effect of RO consumption on resistance of rat testicular cells (TCs) against DNA-damaging effects of the oxidative agents H₂O₂ and DMNQ and on the activity of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD). DNA lesions were detected by conventional and modified comet assay and the activities of GSH-Px and SOD were measured spectrophotometrically. Since TCs represent a mixture of haploid, diploid and tetraploid cells, we used flow cytometry for their differentiation and calculation of DNA-damaging effects of H₂O₂ and DMNQ in cells of different ploidy. The results showed that the oxidative DNA lesions were significantly reduced in TCs from rats administered RO; however, the activity of antioxidant enzymes did not differ in TCs from control and RO-supplemented rats.     

Chemoprotective action of l-(+)-selenomethionine on the modulation of genes involved in oxidative stress and in the UPR pathway

The installation of oxidative process arises from an imbalance between oxidants and antioxidants compounds, in favor of excessive generation of free radicals. This process can affect cellular components, like endoplasmic reticulum (ER). The ER is extremely sensitive to changes in homeostasis, where, on different stimuli, may result in adaptation to survival or induction of apoptosis (unfolded protein response, “UPR” pathway). The oxidative damage caused by endogenous or exogenous agents or a dysregulation of the UPR pathway can lead to adverse conditions, whose chronicity has important implications for the etiology of chronic diseases, including cancer. Therefore, it becomes important to search for chemoprotective agents aiming their role in preventive medicine. Between these substances, selenium’s antioxidant activity seems to be effective in treating diseases which have the oxidative stress as its development. We evaluated the modulating action of l-(+)-selenomethionine (SeMet) in HepG2 cells against cellular stress induced by H₂O₂ through MTT assay, comet assay, and gene expression by qRT-PCR of genes related to oxidative stress, UPR pathway, and apoptosis. In MTT assay, the lower concentrations of SeMet showed a cytoprotective action against the damage caused by H₂O₂. Likewise, it was verified in the comet assay that the concentration of 50 ng/mL reduced the genotoxic damage caused by H₂O₂. SeMet at 50 ng/mL regulated the genes tested in the qRT-PCR, showing an antiapoptotic and antioxidant effect. These results suggest that SeMet positively modulates the genes of oxidative stress and ER, leading to a chemoprotective and antioxidant effect, becoming an alternative in preventive medicine.     

A proposed mechanism of action of textile/Al2O3–TiO2 bimetal oxide nanocomposite as an antimicrobial agent

The textile/Al₂O₃–TiO₂ bimetal oxide nanocomposite has been developed as an antimicrobial agent, but its antimicrobial mechanism has still not been clarified. The Al₂O₃–TiO₂ bimetal oxide nanoparticle has been reported as a radical scavenger. This study focuses on investigating the antimicrobial mechanism of the textile/Al₂O₃–TiO₂ bimetal oxide nanocomposite against Escherichia coli and their interaction with cell envelope biomolecules. L-α-Phosphatidyl ethanolamine (PE) is used as a model of bacteria to investigate the antimicrobial mechanism of this nanocomposite. The antimicrobial activity of the textile/Al₂O₃–TiO₂ bimetal oxide nanocomposite was investigated by using attenuated total reflectance/Fourier transform infrared (ATR-FTIR) and UV–Vis, while the toxicity of this nanocomposite was also examined through tissue culture test against a fibroblast skin cell. The ATR-FTIR used was able to confirm the destroyed cell envelope of the bacteria. The amounts of reactive oxygen species (ROS) were analyzed by UV–Vis spectroscopy and the toxicity of this nanocomposite was also examined by reacting tissue culture against the fibroblast skin cell. Overall, the antimicrobial mechanism of this textile nanocomposite was first by the attachment of this nanocomposite through the attachment of this nanoparticle to the surface of PE (as the model of bacteria) by hydrogen binding, and then nanoparticles can destroy the cell wall of bacteria through oxidation reaction by the produced ROS. Finally, these nanoparticles scavenged the ROS free radical. Therefore, the attached nanoparticles attached on textile can kill bacteria without any ROS free radical remaining in human body. These results suggest that the antimicrobial mechanism of this nanocomposite is mostly different due to the scavenger ability of this nanocomposite and its lower toxicity.     

Cytoprotective effect of white tea against H2O2-induced oxidative stress in vitro

The protective effect of water extracts of white tea (WEWT) on oxidative stress in vitro is investigated. WEWT, like water extracts of green tea (WEGT) and water extracts of Pu-erh tea (WEPT), demonstrates a marked inhibition of the oxidation of liposome, albumin and LDLmodel systems. WEWT protects against H₂O₂-induced cytotoxicity, in a dose-dependent manner. The inhibition of ROS generation and MDA formation by WEWT in H₂O₂-induced Clone 9 cells parallels the effects on cell viability. Moreover, GSH and antioxidant enzymes may play an important role in the protective effect that is associated with H₂O₂-induced oxidative stress. The HPLC-DAD and HPLC–MS/MS analysis, shows that sixteen bioactive compounds are present in WEWT, which may partially account for its protective effect against oxidative insult. These results suggest that the mechanism of the protective actions of WEWT is related to its antioxidant potential and the maintenance of the normal redox status of the cell.     

Cytoprotective effect of fucoxanthin isolated from brown algae <Emphasis Type="Italic">Sargassum siliquastrum</Emphasis> against H<Subscript>2</Subscript>O<Subscript>2</Subscript>-induced cell damage

In this study, the cytoprotective effect of fucoxanthin, which was isolated from Sargassum siliquastrum, against oxidative stress induced DNA damage was investigated. Fucoxanthin, a kind of carotenoid, was pretreated to the medium and the protective effect was evaluated via 2′,7′-dichlorodihydrofluorescein diacetate, 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide, and comet assays. Intracellular reactive oxygen species were over produced by addition of hydrogen peroxide (H₂O₂), but the production was significantly reduced by the treatment with fucoxanthin. The fucoxanthin strongly enhanced cell viability against H₂O₂ induced oxidative damage and the inhibitory effect of cell damage was a dose-dependent manner. Furthermore, a protective effect against oxidative stress-induced cell apoptosis was also demonstrated via nuclear staining with Hoechst dye. These results clearly indicate that fucoxanthin isolated from S. siliquastrum possesses prominent antioxidant activity against H₂O₂-mediated cell damage and which might be a potential therapeutic agent for treating or preventing several diseases implicated with oxidative stress.     

Antioxidant activity of cyanidins isolated from Ogapy (Acanthopanax divaricatus var. albeofructus) fruits in U937 macrophages

Reactive oxygen species (ROS) are continuously produced in aerobic organisms. Overproduction of ROS is known to play a crucial role in the pathogenesis of many cardiovascular diseases, including atherosclerosis and hypertension. Superoxide dismutase (SOD) and catalase (CAT) play critical roles on the removal of excess ROS. In the present study, we investigated the antioxidant activity of cyanidins from ogapy (Acanthopanax divaricatus var. albeofructus, ADA) fruits against oxidative stress in hydrogen peroxide (H₂O₂)-pretreated U937 macrophages, and explored the plausibility of the therapeutic effect of cyanidins on atherosclerosis. As a result, H₂O₂ generation and lipid peroxidation induced by H₂O₂-pretreatment was decreased by the treatment of cyanidins in U937 macrophages. In addition, increased activity of SOD and CAT was shown in H₂O₂-pretreated cells when treated with cyanidins. Overall, the results obtained in this study showed that cyanidin 3-galactoside and cyanidin 3-lathyroside from ADA fruits could protect macrophages against the damaging effects of H₂O₂ treatment.     

Effects of anthocyanins and other phenolics of boysenberry and blackcurrant as inhibitors of oxidative stress and damage to cellular DNA in SH‐SY5Y and HL‐60 cells

There is growing interest both from consumers and researchers in the role that berries play in human health. The objective of this study was to investigate whether anthocyanins and other phenolics present in boysenberries and blackcurrants are effective in protecting cells against the oxidative damage induced by hydrogen peroxide (H₂O₂). The concentrations of polyphenols used were within the human physiological range. The data showed that SH‐SY5Y human neuroblastoma cells were protected against H₂O₂‐induced toxicity by the anthocyanins and phenolic fractions. The concurrent addition of either fractions of these berries with H₂O₂ significantly inhibited the increase in intracellular reactive oxygen species (ROS) production. Pre‐incubation of cells with the same concentrations had no effect on the ROS level—a result that may be due to the metabolic conversion to inactive compounds. Anthocyanins and phenolic fractions of blackcurrant were better at protecting DNA of HL‐60 human promyelocytic cells from damage than similar fractions from boysenberry. The phenolic extract of blackcurrant demonstrated the highest protective effect against H₂O₂‐induced neurotoxicity, oxidative stress and DNA damage and may be a good candidate for inclusion into a processed functional food. Copyright © 2006 Society of Chemical Industry     

Effect of polyphenolic compounds from Coriandrum sativum on H2O2-induced oxidative stress in human lymphocytes

Polyphenolic compounds are widely distributed in plants and known to be excellent antioxidants in vitro. They have the capacity to reduce free-radical formation by scavenging free radicals and protecting antioxidant defences. The present study evaluated the antioxidant potencies of polyphenolic compounds from a spice, Coriandrum sativum against hydrogen peroxide-induced oxidative damage in human lymphocytes. Pretreatment with polyphenolic rich fractions protected human lymphocytes against H₂O₂-induced oxidative damage. H₂O₂ treatment significantly decreased the activities of antioxidant enzymes, such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-S-transferase and caused decreased glutathione content and increased thiobarbituric acid-reacting substances (TBARS). Treatment with polyphenolic fractions (50 μg/ml) increased the activities of antioxidant enzymes and glutathione content and reduced the levels of TBARS significantly. Observed reduction in the level of lipid peroxides showed a decreased tendency of peroxidative damage. We conclude that, under these experimental conditions, polyphenolic compounds effectively suppress hydrogen peroxide-induced oxidative stress.     

葡萄籽粗多糖的体内外抗氧化作用

为了研究葡萄籽粗多糖(crude polysaccharides from grape seeds,GSCPs)体外抗氧化作用及对秀丽隐杆线虫的体内抗氧化作用,采用水提醇沉法提取GSCPs,检测GSCPs对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基和羟自由基的清除作用及对DNA氧化损伤的抑制作用;建立RAW 264.7巨噬细胞氧化损伤模型,在细胞水平探讨GSCPs的抗氧化能力;同时利用秀丽隐杆线虫研究GSCPs的体内抗氧化功能。体外实验结果表明:GSCPs可有效清除自由基,抑制DNA的氧化损伤,质量浓度为0.4mg/mL的GSCPs对DPPH自由基的清除率达84%,对羟自由基清除率为89%;GSCPs可以下调H₂O₂诱导的RAW 264.7巨噬细胞活性氧(reactive oxygen species,ROS)水平,正向调节细胞内超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性,质量浓度为0.2mg/mL的GSCPs处理组可使细胞内SOD活性从H₂O₂处理组的81.1%升至96.3%,GSH-Px活性从H₂O₂处理组的92.1%升至99.6%。此外,GSCPs可延长秀丽隐杆线虫寿命,提高其对抗急性氧化应激的能力,有效清除秀丽隐杆线虫体内的ROS。质量浓度为0.8mg/mL GSCPs处理组秀丽隐杆线虫的平均寿命较对照组延长27.67%,将急性氧化应激的秀丽隐杆线虫平均存活时间延长33.58%,秀丽隐杆线虫体内ROS生成量较对照组可降低56.33%。因此,葡萄籽粗多糖在体内外均表现出良好的抗氧化性,可用于抗氧化功能产品的开发。     

Protective effect of two edible mushrooms against oxidative cell damage and their phenolic composition

The aim of the study was to investigate phenolic composition, antioxidative, protective and cytotoxic effects of Pleurotus eryngii and Auricularia auricula-judae. Analysis of phenolic compounds in these edible mushrooms species has been carried out by high-performance liquid chromatography (HPLC). Protective effect of these mushrooms on H₂O₂ induced oxidative cell damage was determined by using MTT (3-(4,5-Dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide, a yellow tetrazole) assay. Antioxidant activities of the mushrooms extracts were evaluated by using complementary in vitro assays. In addition, the measurement of total antioxidant compounds in the extracts was carried out. All the extracts exhibited protective effect against H₂O₂ induced oxidative cell damage but the highest activity was observed for A. auricula-judae aqueous extract (89.5 ± 1.8% cell viability at 0.1 mg/ml). P. eryngii methanolic extract showed the highest ferrous iron chelating ability (IC₅₀ = 0.42 ± 0.03 mg/ml). A. auricula-judae extracts (at concentration of 0.025–0.100 mg/ml) were not toxic to baby hamster kidney fibroblast cell line (BHK 21). These results suggest that these mushrooms may be used as a potential source of natural antioxidants for food supplementation or in the development of nutraceuticals.     

Glycine betaine improves oxidative stress tolerance and biocontrol efficacy of the antagonistic yeast Cystofilobasidium infirmominiatum

The effect of H₂O₂-induced oxidative stress on the viability of the yeast antagonist, Cystofilobasidium infirmominiatum, as well as the effect of exogenous glycine betaine (GB) on yeast viability under oxidative stress, was determined. GB treatment improved the tolerance of C. infirmominiatum to oxidative stress. Compared to untreated control yeast cells, GB-treated cells showed less accumulation of reactive oxygen species (ROS) and a lower level of protein oxidation in response to oxidative stress. Additionally, GB-treated yeast exhibited greater biocontrol activity against Penicillium expansum and a faster growth in wounds of apple fruits stored at 25 °C compared to the performance of untreated yeast. The activities of antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPX) of C. infirmominiatum were elevated by GB treatment. Results indicate that the elicitation of antioxidant response by GB may contribute to improvements in oxidative stress tolerance, population growth in apple wounds, and biocontrol activity of C. infirmominiatum.     

Antioxidant activities of phlorotannins purified from Ecklonia cava on free radical scavenging using ESR and H2O2-mediated DNA damage

The potential antioxidant activities of three phlorotannins (phloroglucinol, eckol and dieckol) purified from Ecklonia cava collected in Jeju Island were investigated to evaluate their potential value as the natural products for foods or cosmetic application. In this study, antioxidant activities were measured by electron spin resonance spectrometry (ESR) technique for scavenging effects of free radicals such as 1,1-diphenyl-2-picrylhydrazyl (DPPH), alkyl, hydroxyl (HO^•) and superoxide anion radical (O₂ ^•−) and by comet assay for protecting effects against H₂O₂-mediated DNA damage. The results show that all the phlorotannins have the potential DPPH, alkyl, hydroxyl and superoxide radical scavenging activities. Especially, eckol samples scavenged around 93% of DPPH at 0.25, 0.5, 1 mg/mL of concentrations and were higher than the other phlorotannins, such as phloroglucinol and dieckol samples. Also, protecting effects of the phlorotannins against H₂O₂-mediated DNA damage increased with increased concentrations of the samples in the L5178 mouse T-cell lymphoma cell lines (L5178Y-R). In conclusion, these results suggest that the three phlorotannins purified from E. cava have the potential inhibitory effect on H₂O₂-mediated DNA damage and harmful free radicals and can be used as antioxidants in cosmetic, foods and drug industry.     

Carnosic acid protects against ROS/RNS-induced protein damage and upregulates HO-1 expression in RAW264.7 macrophages

The overproduction of reactive oxygen species (ROS) and reactive nitrogen species (RNS) is a deleterious process that can be an important mediator of damage to cell structures including protein, lipid and DNA, and consequently leads to various disease states and senescence. In the present study, the protective effects of carnosic acid (CA) on ROS/RNS-induced protein damages were examined. CA dose-dependently decreased the fragmentation, carbonylation, and nitration of bovine serum albumin (BSA) in AAPH and Hemin/nitrite/H₂O₂ incubation systems, respectively. Moreover, CA effectively attenuated protein carbonylations in the radical-treated RAW264.7 cells. Furthermore, after pretreatments of RAW264.7 macrophages with CA, the generation of ROS and NO induced by AAPH and H₂O₂ or LPS were significantly suppressed, and heme oxygenase-1 (HO-1) protein expression was time- and dose-dependently upregulated. Hence, our results indicated that CA might be beneficial for cellular proteins in oxidative stress or inflammation conditions by alleviating ROS/RNS generation and inducing the expression of antioxidant enzyme HO-1.     

Protective effect of Centella asiatica extract and powder on oxidative stress in rats

The effect of Centella asiatica extract and powder in reducing oxidative stress in SpraqueDawley rats was evaluated. Lipid peroxidation was monitored by measuring malonaldehyde (MDA) level in blood. Activities of free radical-scavenging enzymes (superoxide dismutase and catalase) were determined using H₂O₂ decomposition and nitrobluetetrazolium reduction, respectively. Results showed that administration of H₂O₂ (0.1%) in drinking water of the rats, for 25 weeks, increased the malonaldehyde levels in erythrocytes of all the rats. However, rats receiving C. asiatica extract, powder and α-tocopherol had lower MDA levels than did the other rats, which indicates, decrease lipid peroxidation in these rats. Increase in catalase activity of the rats appears to be a response to H₂O₂ accumulation. The decrease in the activity of superoxide dismutase in C. asiatica- and α-tocopherol supplemented rats suggested a lower requirement for the enzyme and this indicates the protective effect of the plant in combating oxidative stress undergone by the rats. Results revealed that C. asiatica extract and powder may ameliorate H₂O₂-induced oxidative stress by decreasing lipid peroxidation via alteration of the antioxidant defence system of the rats.     

Free radical scavenging activity and comparative proteomic analysis of antioxidative protein against H2O2-induced oxidative stress in neuronal cells

Artemisia annua was enzymatically hydrolyzed by five proteases and seven carbohydrases. All enzymatic extracts scavenged DPPH, hydroxyl and alkyl radicals. Especially, the Protamex among the various proteases and Maltogenase among the various carbohydrases extracts exhibited the highest scavenging activity on hydroxyl radical. The extracts of A. annua clearly reduced neuronal cell death from H₂O₂-induced damage. In addition, a proteomic analysis, two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionisation-time of flight/time of flight (MALDI-TOF/TOF) was used to identify the proteins of the neuronal cells whose expressions were or were not altered by the treatment of the Maltogenase extracts which showed the highest hydroxyl radical scavenging activity among all enzymatic extracts for 24 h. The protein characterisation revealed that translation elongation factor Tu (EF-Tu), Immunoglobulin E (IgE) and voltage-dependent anion channel 1 (VDAC-1) were involved in the cell survival effects against H₂O₂-induced apoptosis. These results suggest that EF-Tu, IgE and VDAC-1 have an important role in the reduction of neuronal apoptosis by oxidative stress, and the enzymatic extracts of A. annua shows potent antioxidative activities by regulating EF-Tu, IgE and VDAC-1.     

Kinetics of protein physicochemical changes induced by heating in meat using mimetic models: (2) Effects of fibre type,peroxides and antioxidants

Heating-induced changes in meat proteins were investigated using models made of aqueous suspensions of myofibrils according to muscle fibre types and cellular compounds (oxidants and antioxidants). These changes were evaluated by measurements of carbonyl groups and protein surface hydrophobicity. Model results were compared to trial results obtained on pork meat (M. Longissimus dorsi) heated under the same conditions (45 and 75 °C, from 5 to 120 min). Myofibrillar proteins from α-white fibres were more sensitive to oxidation and thermal denaturation than those from β-red fibres. At 45 °C, there were negligible differences due to peroxide or antioxidant types. At 75 °C, organic peroxides (ROOH) were less oxidative than hydrogen peroxide (H₂O₂), and antioxidant enzymes were less efficient than vitamin E and carnosine at protecting proteins against oxidation. Protein oxidation observed in meat is lower than in the mimetic models and the increase in hydrophobicity remained limited in meat.