Antioxidant effect derived from bioaccessible fractions of fruit beverages against H2O2-induced oxidative stress in Caco-2 cells

This work evaluates the effect of bioaccessible fractions from fruit beverages against oxidative stress (OS) in Caco-2 cells. A fruit beverage (grape + orange + apricot) (with/without milk and/or iron/zinc) was subjected to in vitro gastrointestinal digestion, and bioaccessible fractions were incubated with Caco-2 cell cultures. Following preincubation, OS was induced with 5 mM H₂O₂. Intracellular reactive oxygen species (ROS), mitochondrial potential (Δψ_m), mitochondrial metabolism (MTT test), intracellular reduced glutathione (GSH) and superoxide dismutase activity (SOD) were measured. The data evidenced viable cultures with increased mitochondrial metabolism and GSH-Rd activities, without alteration in SOD activity. Accordingly, more preserved mitochondrial integrity was also evidenced, allowing the action of antioxidant systems in preincubated cultures. Based on these data, we can conclude that a cytoprotective effect is derived from bioaccessible fractions of fruit beverages, though this effect failed to prevent intracellular ROS accumulation in Caco-2 cell cultures exposed to 5 mM H₂O₂.     

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.     

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.     

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.     

英国红芸豆抗氧化肽组分对H2O2诱导PC12细胞氧化应激损伤的保护作用

为了探究英国红芸豆抗氧化肽组分的抗氧化作用,采用超滤、葡聚糖凝胶G-15层析技术对碱性蛋白酶酶解英国红芸豆抗氧化蛋白质制备的抗氧化产物进行分级,研究抗氧化活性较高的组分对H₂O₂损伤的PC12细胞的影响。结果表明:抗氧化肽组分BRKBAPC-2能缓解H₂O₂对PC12造成的细胞生长抑制作用,随着肽浓度的提高,细胞内活性氧（ROS）水平,丙二醛（MDA）含量及乳酸脱氢酶（LDH）胞外活力均有所降低,超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活力以及还原型谷胱甘肽（GSH）含量均具有所提高,尤其BRKBAPC-2浓度为200 μg/mL时,ROS水平为32.53%,MDA含量为0.96 nmol/μg prot,LDH胞外活力为202.12 U/L,与模型组相比,均极显著降低（P<0.01）;SOD活力为（555.48±3.64）U/mg prot,CAT活力为（14.77±1.30）U/mL,GSH含量为（140.88±8.19）μmol/g prot,与模型组相比,均极显著提高（P<0.01）,此外,其可抑制线粒体膜电位的降低,改善细胞周期的阻滞情况,阻止细胞凋亡关键酶Caspase-3和Caspase-9的激活。可见,抗氧化肽组分BRKBAPC-2对H₂O₂引起的PC12细胞氧化损伤具有一定的保护作用。     

Antioxidant properties of peptide fractions from silver carp (Hypophthalmichthys molitrix) processing by-product protein hydrolysates evaluated by electron spin resonance spectrometry

Silver carp processing by-product protein is usually discarded as an industrial solid waste. In this study the protein was recovered using a pH-shift method, after which seven commercial proteases were separately employed to prepare antioxidative hydrolysates. Among the hydrolysates, pepsin hydrolysates, which had the highest free radical-scavenging activity, were further separated into five peptide fractions, SCPH-I (>10 kDa), SCPH-II (5–10 kDa), SCPH-III (3–5 kDa), SCPH-IV (1–3 kDa), and SCPH-V (<1 kDa), by using ultrafiltration. The antioxidative properties of the peptide fractions were investigated, using a free radical-scavenging assay, by electron spin resonance. The results show that SCPH-V had the highest scavenging effects on DPPH (1,1-diphenyl-2-picrylhydrazyl), hydroxyl and superoxide anion radicals. SCPH-V had potent antioxidant activity in the prevention of the peroxidation of linoleic acid and alleviation of H₂O₂-induced oxidative stress in human intestinal epithelial caco-2 cells. The results indicated that the antioxidant capacity of silver carp by-product hydrolysates could be enhanced by ultrafiltration.     

婴幼儿配方奶粉对人结肠腺癌系Caco-2细胞和SD大鼠腹腔巨噬细胞自由基水平的影响

为探究婴幼儿配方奶粉对消化道上皮细胞及黏膜免疫系统自由基水平调节能力的影响,随机选取5种市售一段婴幼儿配方奶粉,以SD大鼠腹腔巨噬细胞和人结肠腺癌系Caco-2细胞为检测模型,测定配方奶粉对细胞增值率影响的质量浓度范围,并采用经部分修改的细胞内抗氧化活性测定方法(CAA)评价配方奶粉的胞内抗氧化能力.在无外源性自由基诱...     

Carotenoid compositions of coloured tomato cultivars and contribution to antioxidant activities and protection against H2O2-induced cell death in H9c2

The carotenoid compositions, antioxidant activities and the potential cardio-protective role of 13 tomato cultivars with distinct colour were studied. Colour coordinates were evaluated by colorimeter and the carotenoid compositions were analysed by UPLC. Red tomatoes had the highest total carotenoid contents (TCC) and antioxidant activities, followed by purple, orange, pink and yellow ones. The TCC were 120.5–278.0 μg/g DW, and the antioxidant activities were 21.32–40.07 μmol TE/g DW (PCL), 64.42–89.98% (DPPH) and 10.47–13.76 μmol TE/g DW (ORAC), respectively. The lipophilic extracts were also found to prevent cell death in a cell-based model system using cardiac H9c2 cells and H₂O₂, via attenuation of the caspase-3 and matrix metalloproteinase-2 activities. The extracts of different tomatoes showed strong but different antioxidant activities. Roles of total and individual carotenoids in the antioxidant activities were studied and lycopene showed the highest correlation. Results of this study can be used to guide the development of new tomato cultivars and functional foods, and benefit the consumers.     

Hepatoprotective peptides purified from Corbicula fluminea and its effect against ethanol-induced LO2 cells injury

This study was focused on the purification, identification and mechanism of hepatoprotective peptides from Corbicula fluminea by using Sephadex G-15 chromatography, UPLC–MS/MS, oxygen radical absorbance capacity (ORAC) assay, cell experiment and molecular docking. Six identified peptides were synthesised chemically and subjected to evaluate hepatoprotective effect. ORAC value and hepaprotective effect against ethanol-induced LO2 cell injury in vitro were determined. The results showed that Tyr-Phe-Leu-Pro (YP-4) and Leu-Val-Tyr-Pro (LP-4) exhibited the strongest antioxidant activity and significantly increased the viability of ethanol-injured LO2 cells. These two peptides significantly reduced ROS levels and efficiently inhibited the decrease of mitochondrial membrane potential in ethanol-injured LO2 cells. Molecular docking revealed that YP-4 and LP-4 possess potential inhibitory activity on CYP2E1 and thus reduce ethanol-induced oxidative stress. It is suggested that YP-4 and LP-4 have good hepatoprotective effect against alcoholic liver injury.     

金银花叶黄酮体外抗氧化能力及对H2O2诱导RAW264.7巨噬细胞损伤的保护作用

目的：研究金银花叶黄酮的体外抗氧化能力和对H2O2诱导RAW264.7巨噬细胞损伤的保护作用。方法：金 银花叶粉经提取纯化后得到金银花叶黄酮粉,以抗坏血酸为阳性对照,测定金银花叶黄酮的总还原力,对羟自由 基、超氧阴离子自由基及1,1-二苯基-2-三硝基苯肼自由基的清除能力。体外培养RAW264.7巨噬细胞,实验分为空 白组、模型组、对照组和金银花叶黄酮低、中、高剂量组,用H2O2诱导损伤RAW264.7细胞,噻唑蓝法测定细胞存 活率,试剂盒法测定细胞和细胞培养液中丙二醛、谷胱甘肽含量及超氧化物歧化酶、乳酸脱氢酶活力。结果：金银 花叶黄酮的总还原力及对各自由基的清除能力较强,并在足够质量浓度下等同于对照品抗坏血酸。金银花叶黄酮呈 剂量依赖性保护H2O2引起的RAW264.7细胞的损伤,降低细胞及细胞培养液中丙二醛含量,提高超氧化物歧化酶活力 及谷胱甘肽含量,提高细胞内乳酸脱氢酶活力。结论：金银花叶黄酮抗氧化能力较强,可修复H2O2诱导的RAW264.7 巨噬细胞的损伤,其作用可能与调节细胞氧化还原系统、清除自由基、提高细胞内抗氧化酶系的活力有关。     

Protective effects of the crude extracts from yam (Dioscorea alata) peel on tert-butylhydroperoxide-induced oxidative stress in mouse liver cells

Researchers have shown that yam extracts contain antioxidative activity; however, there are few reports regarding the antioxidant activities of yam peel. The effects of water and 50% ethanolic extracts from Darsan yam (Dioscorea alata) peel on the oxidative status of tert-butylhydroperoxide (t-BHP)-treated mouse Hepa 1–6 and FL83B liver cell lines were investigated. The cytosols were analysed for H₂O₂ and malondialdehyde (MDA) levels and antioxidative enzymes activities, including superoxide dismutase, glutathione peroxidase (GPx) and catalase activities. Both water and 50% ethanolic extracts from yam peel did not affect cellular MDA level in t-BHP-treated cells, but they altered the level of H₂O₂. Water extract from yam peel amplified the t-BHP-induced cytotoxicity in Hepa 1–6 whilst the ethanolic extract showed protection in FL83B cells. GPx activity might play an important role in the protective effect associated with t-BHP-induced oxidative stress.     

Antioxidant activity of sugarcane molasses against 2,2′-azobis(2-amidinopropane) dihydrochloride-induced peroxyl radicals

Sugarcane molasses is a rich source of antioxidant materials with peroxyl radical scavenging effects. To explore the potent antioxidant activity of sugarcane molasses against 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced peroxyl radicals, 7 methanolic fractions of sugarcane molasses (F1–F7) were separated via bioassay-guided fractionation and evaluated by oxygen radical absorbance capacity (ORAC), cellular antioxidant activity (CAA), and oxidative DNA damage protective activity assays. The ORAC values of sugarcane molasses fractions ranged from 4399 to 6266 μmol TE/g, whilst the EC₅₀ values for CAA ranged from 3.7 to 5.9 μg/ml. Moreover, it was found that sugarcane molasses fractions, particularly F6 and F7, could protect against oxidative DNA damage caused by peroxyl radicals at an effective concentration of 100 μg/ml. Ten phenolic constituents were identified in the fractions, including known antioxidative compounds, viz., schaftoside, isoschaftoside, ferulic acid, p-coumaric acid, and p-hydroxybenzaldehyde.     

Napiergrass (Pennisetum purpureum S.) protects oxidative damage of biomolecules and modulates antioxidant enzyme activity

The effects of water extract of napiergrass (Pennisetum purpureum S.) (WEN) on oxidative damage of biomolecules and modulation of antioxidant enzyme activity were investigated. The results showed that WEN displayed marked free radical scavenging, reducing power, as well as ferrous ions chelating effects. WEN has a dose-dependent response for protective action on oxidation of phospholipid, deoxyribose and low-density lipoprotein (LDL) in the range of 0–0.5 mg/ml, indicating that WEN had in vitro protective action on oxidative damage of biomolecules. Oxidative stress induced by H₂O₂ significantly decreased the viability of BNL cells. However, addition of WEN in the medium protected cells from H₂O₂-induced cytotoxicity. Furthermore, treatment of cells with WEN in the range of 0–0.2 mg/ml displayed protective effect from H₂O₂ induced oxidation in a concentration dependent manner. With respect to the effect of WEN on antioxidant enzymes, the results showed the WEN at 0.2 mg/ml enhanced activities of glutathione peroxidase (GPX), glutathione reductase (GR), glutathione transferase (GST) and catalase (CAT) in BNL cells by 2.93-, 35.8-, 4.23-, and 2.78-fold, respectively, compared to the control; WEN increased the GSH content by 3.2-fold, implying that WEN may up-regulate the levels of GSH and antioxidant enzymes in BNL cells. WEN scavenged NO generated by a NO donor, sodium nitroprusside (SNP) and suppressed NO production in lipopolysaccharide (LPS)-activated macrophage RAW 264.7 cells. The determination of ascorbic acid and total anthocyanins as well as HPLC analysis revealed that ascorbic acid, rutin, epicatechin, anthocyanins, p-coumaric acid, quercetin and catechin were present in WEN, which function as in vitro antioxidants by virtue of their ability to scavenge ROS and RNS. Overall, the results obtained showed that WEN is rich in antioxidant components and they can serve as an excellent potential for use as a natural phytochemicals source.     

Cranberries and Wild Blueberries Treated with Gastrointestinal Enzymes Positively Modify Glutathione Mechanisms in Caco‐2 Cells In Vitro

Beneficial health effects of cranberries (CBs) and wild blueberries (BBs), such as reduced levels of oxidative stress, have been demonstrated in feeding studies. These Vaccinium berries contain high levels of flavonoids; however, the bioavailability of flavonoids is generally low. We investigated the in vitro effects of these berries on intestinal cells, focusing on mitigating oxidative stress and associated reactive oxygen species (ROS). First, we simulated the passage of CB and BB through the gastrointestinal (GI) tract by treating berry homogenates to a battery of digestive enzymes. Then, Caco‐2 cells, a model of small intestine epithelial uptake, were exposed to these homogenates for 60 min. Using a cell‐free assay, we found that the antioxidant activity in CB homogenates was not affected by these enzymes, but that BB homogenates treated with gut enzymes had 43% lower free‐radical quenching activity (P < 0.05). However, both of the enzyme‐treated homogenates were still able to counteract the ROS‐generating ability of H₂O₂ added exogenously to Caco‐2 cells. Berry homogenates also increased mitochondrial metabolic rates at 60 min posttreatment, as measured by MTT assays. Enzyme‐treated CB (but not BB) homogenates increased the levels of reduced glutathione (GSH) relative to oxidized glutathione (GSSG), a critical indicator of the cellular redox state (P < 0.05). Our data suggest that CBs do not lose their antioxidant ability when passing through the GI tract, and specifically, digested CB may serve to enhance cytoprotective effects in intestinal cells by reducing potential damage caused by free radicals and ROS derived from other food sources.     

Antioxidant and antiproliferative activities of mango (Mangifera indica L.) flesh and peel

The antioxidant and antiproliferative properties of flesh and peel of mango (Mangifera indica L.) were investigated. The cytoprotective effect of mango flesh and peel extracts on oxidative damage induced by H₂O₂ in a human hepatoma cell line, HepG2, were determined, and the underlying mechanism was examined by a single-cell electrophoresis assay (comet assay). Treatment of HepG2 cell with mango peel extract prior to oxidative stress was found to inhibit DNA damage. The free radical scavenging activities of mango flesh and peel extracts were evaluated by electron spin resonance (ESR). The mango peel extract exhibited stronger free radical scavenging ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and alkyl radicals than mango flesh extract, regardless of ripeness. Similarly, peel extract exhibited significant antiproliferative effect against all tested cancer cell lines, compared to that of flesh extract, in a dose-dependent manner. The result also showed that the antiproliferative activity of mango flesh and peel extracts correlated with their phenolic and flavonoid contents. Thus, mango peel, a major by-product obtained during the processing of mango product, exhibited good antioxidant activity and may serve as a potential source of phenolics with anticancer activity.     

鲈鱼抗氧化肽的稳定性分析及其对细胞氧化损伤的保护作用

目的:探究鲈鱼蛋白酶解物（Lateolabrax maculatus protein hydrolysates,LPH）的体外抗氧化活性及稳定性。方法:采用化学实验测定LPH的抗氧化活性以及分析pH、温度、金属离子（K+、Ca2+、Cu2+、Zn2+）和模拟胃肠道消化对其稳定性的影响,并利用过氧化氢诱导的Caco-2细胞氧化损伤模型探究LPH对细胞氧化损伤的保护作用。结果:LPH具有较强的自由基清除活性,其DPPH和ABTS+自由基清除能力的IC₅₀值分别为2.13 mg/mL和31.53 μg/mL。LPH对pH（2.0~12.0）、温度（25~100 ℃）和K+（0.25~2 mmol/L）稳定,0.25~2 mmol/L的Ca2+和Cu2+能够提升其DPPH自由基清除率,而1 mmol/L以上的Zn2+会降低DPPH自由基清除率。此外,LPH具有良好的胃肠道稳定性。在过氧化氢诱导的Caco-2细胞模型中,LPH能够将氧化损伤的细胞存活率由58.02%显著增加至83.40%（P<0.05）,显著抑制细胞内活性氧的释放（P<0.05）并恢复细胞线粒体膜电位至正常水平。此外,LPH能够显著抑制细胞上清中乳酸脱氢酶水平至模型组的19.34%（P<0.05）并显著增加细胞内超氧化物歧化酶和过氧化氢酶活力（P<0.05）。结论:LPH具有较好的抗氧化活性及稳定性,在食品保健领域具有良好的应用前景。     

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.     

Antiproliferative effects of cherry juice and wine in Chinese hamster lung fibroblast cells and their phenolic constituents and antioxidant activities

Cherries are good sources of bioactive phenolic compounds that are widely considered to be potentially healthy. Here we investigated the protective activities of juice and wine products of tart and sweet cherries and their constituent anthocyanins (e.g., cyanidin 3-glucoside and cyanidin 3-rutinoside) against oxidative stress induced by hydrogen peroxide (H₂O₂) in Chinese hamster lung fibroblasts (V79-4). Total phenolics in the cherry juices and wines were 56.7–86.8 mg of gallic acid equivalents (GAE)/l and 79.4–149 mg GAE/l, respectively. Total anthocyanins in the cherry juices and wines were 7.9–50.1 mg of cyanidin 3-glucoside equivalents (CGE)/l and 29.6–63.4 mg CGE/l, respectively. Both cherry juices and wines exerted protective effects against oxidative stress induced by H₂O₂ on V79-4 cells and also enhanced the activities of antioxidative enzymes, such as superoxide dismutase and catalase, in a dose-dependent manner. The protection of V79-4 cells from oxidative stress by phenolics was mainly attributable to anthocyanins. The positive correlation between the protective effects against oxidative stress in V79-4 cells and the antioxidant enzyme activities was stronger for cyanidin 3-glucoside than for cyanidin 3-rutinoside.     

Antioxidant activity and hepatoprotective effect of Schizandra chinensis Baill. Extracts containing active components in alcohol-induced HepG2 cells

The antioxidant activities and hepatoprotective effects of Schizandra chinensis Baill. extracts (SCE) were evaluated. The contents of the active components schisandrin, schisandrin C, gomisin A, and gomisin N in SCE were 8.802±1.390, 1.011±0.203, 0.954±0.191, and 3.351±0.829 mg/g, respectively. The antioxidant activity of SCE was measured based on the DPPH free radical scavenging activity and the superoxide dismutase (SOD)-like activity. The IC₅₀ values for the DPPH radical scavenging activity and SOD-like activity were 18.1 and 44.2mg/mL, respectively. The protective effect of SCE against alcohol-induced oxidative injury in HepG2 cells was investigated. Cells pretreated with SCE (100–400 μg/mL) showed an increased resistance to oxidative stress in a dose-dependent manner. Expressions of Bcl-2 and pro-caspase-3 proteins were induced by SCE in HepG2 cells. SCE can be useful for management of antioxidant and hepatoprotective effects.