The Cytoprotective Effect of Petalonia binghamiae Methanol Extract against Oxidative Stress in C2C12 Myoblasts: Mediation by Upregulation of Heme Oxygenase-1 and Nuclear Factor-Erythroid 2 Related Factor 2

This study was designed to examine the protective effects of the marine brown algae Petalonia binghamiae against oxidative stress-induced cellular damage and to elucidate the underlying mechanisms. P. binghamiae methanol extract (PBME) prevented hydrogen peroxide (H₂O₂)-induced growth inhibition and exhibited scavenging activity against intracellular reactive oxygen species (ROS) induced by H₂O₂ in mouse-derived C2C12 myoblasts. PBME also significantly attenuated H₂O₂-induced comet tail formation in a comet assay, histone γH2A.X phosphorylation, and annexin V-positive cells, suggesting that PBME prevented H₂O₂-induced cellular DNA damage and apoptotic cell death. Furthermore, PBME increased the levels of heme oxygenase-1 (HO-1), a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2 related factor 2 (Nrf2). However, zinc protoporphyrin IX, a HO-1 competitive inhibitor, significantly abolished the protective effects of PBME on H₂O₂-induced ROS generation, growth inhibition, and apoptosis. Collectively, these results demonstrate that PBME augments the antioxidant defense capacity through activation of the Nrf2/HO-1 pathway.     

An Ethanol Extract Derived from Bonnemaisonia hamifera Scavenges Ultraviolet B (UVB) Radiation-Induced Reactive Oxygen Species and Attenuates UVB-Induced Cell Damage in Human Keratinocytes

The present study investigated the photoprotective properties of an ethanol extract derived from the red alga Bonnemaisonia hamifera against ultraviolet B (UVB)-induced cell damage in human HaCaT keratinocytes. The Bonnemaisonia hamifera ethanol extract (BHE) scavenged the superoxide anion generated by the xanthine/xanthine oxidase system and the hydroxyl radical generated by the Fenton reaction (FeSO₄ + H₂O₂), both of which were detected by using electron spin resonance spectrometry. In addition, BHE exhibited scavenging activity against the 1,1-diphenyl-2-picrylhydrazyl radical and intracellular reactive oxygen species (ROS) that were induced by either hydrogen peroxide or UVB radiation. BHE reduced UVB-induced apoptosis, as shown by decreased apoptotic body formation and DNA fragmentation. BHE also attenuated DNA damage and the elevated levels of 8-isoprostane and protein carbonyls resulting from UVB-mediated oxidative stress. Furthermore, BHE absorbed electromagnetic radiation in the UVB range (280–320 nm). These results suggest that BHE protects human HaCaT keratinocytes against UVB-induced oxidative damage by scavenging ROS and absorbing UVB photons, thereby reducing injury to cellular components.     

Antioxidative and Anticancer Activity of Extracts of Cherry (<Emphasis Type="Italic">Prunus serrulata</Emphasis> var. <Emphasis Type="Italic">spontanea</Emphasis>) Blossoms

Organic solvent (methanol, ethanol, and acetone) extracts and water extracts of cherry (Prunus serrulata var. spontanea) blossoms were prepared, and antioxidant activities of the extracts were evaluated. Methanolic CBE (100 μg/ml) showed the highest total phenol content (104.30 μM), radical scavenging activity (34.2%), and reducing power (0.391). The effect of CBE on DNA damage induced by H₂O₂ in human leukocytes was evaluated by Comet assay. All CBE was a potent dose dependent inhibitor of DNA damage induced by 200 μM of H₂O₂, methanolic CBE showed the most strong inhibition activity. The methanolic CBE of 500 μg/ml showed 38.8% inhibition against growth of human colon cancer cell line HT-29. These results indicated that cherry blossoms could provide valuable bioactive materials.     

A Search for Hepatoprotective Activity of Fruit Extract of Mangifera indica L. Against Oxidative Stress Cytotoxicity

Mango (Mangifera indica L.) and their components are commonly used in folk medicine for many curative effects. The protective effects of different concentrations of aqueous extract of Mangifera indica L. fruit (Mango Extract) (20, 50 and 100 μg/ml) and also gallic acid (100 μM) as a pure compound in the extract were examined against oxidative stress toxicity induced by cumene hydroperoxide (CHP) in isolated rat hepatocytes. The extracts and gallic acid (100 μM) protected the hepatocyte against all oxidative stress markers including cell lysis, ROS generation, lipid peroxidation, glutathione depletion, mitochondrial membrane potential decrease, lysosomal membrane oxidative damage and cellular proteolysis. Mango Extracts (20, 50 and 100 μg/ml) were more effective than gallic acid (100 μM) in protecting hepatocytes against CHP induced lipid peroxidation. On the other hand gallic acid (100 μM) acted more effective than Mango Extracts (20, 50 and 100 μg/ml) at preventing lysosomal membrane damage. In addition H₂O₂ scavenging effect of all extracts were determined in hepatocytes and compared with gallic acid (100 μM). There were no significance differences (P<0.05) between all plant extracts and gallic acid (100 μM) in H₂O₂ scavenging activity. These results suggest a hepatoprotective role for Mango Extract against liver injury associated with oxidative stress.     

Comparative Study on the Chemical Composition,Antioxidant Properties and Hypoglycaemic Activities of Two <Emphasis Type="Italic">Capsicum annuum</Emphasis> L. Cultivars (<Emphasis Type="Italic">Acuminatum</Emphasis> small and <Emphasis Type="Italic">Cerasiferum</Emphasis>)

The present study aimed to evaluate for the first time the phenols, flavonoids, carotenoids, capsaicin and dihydrocapsaicin content and the antioxidant and hypoglycemic properties of Capsicum annuum var. acuminatum small and C. annuum var. cerasiferum air-dried fruits. The ethanol extract of C. annuum var. acuminatum small, characterized by the major content of total poliphenols, flavonoids, carotenoids and capsaicinoids, showed the highest radical scavenging activity (IC₅₀ of 152.9 μg/ml). On the contrary, C. annuum var. cerasiferum showed a significant antioxidant activity evaluated by the β-carotene bleaching test (IC₅₀ of 3.1 μg/ml). The lipophilic fraction of both C. annuum var. acuminatum and C. annuum var. cerasiferum exhibited an interesting and selective inhibitory activity against α-amylase (IC₅₀ of 6.9 and 20.1 μg/ml, respectively).     

Antioxidant Activity and Phenylpropanoids of Phlomis lychnitis L.: A Traditional Herbal Tea

Phlomis lychnitis L. (Lamiaceae) is consumed as a traditional herbal tea in Spain. The antioxidant-protective effects as well as its phytoconstituents have never been established. The ability of the methanolic extract to protect cells from oxidative stress was evaluated in rat pheochromocytoma cells (PC12) using hydrogen peroxide as toxic agent. The viability of PC12 cells pre-treated with the methanolic extract of Phlomis lychnitis, determined by the MTT and LDH assays, was significantly improved at the highest dose (p?<?0.01). The antioxidant activity was confirmed evaluating the capacity of the plant to scavenge ABTS, DPPH, O₂ · ^? radicals and to inhibit XO. Bioassay guided fractionation led to antioxidant compounds. Qualitative HPLC/DAD/ESI/MS analysis reported phenylpropanoids, verbascoside being the major antioxidant constituent.     

Increased Anti-oxidative Potency of Garlic by Spontaneous Short-term Fermentation

Fundamental anti-oxidative properties of 80% ethanol extract from garlic fermented for the relatively short period of time (40 days at 60–70°C, 85–95% relative humidity) were examined. Superoxide dismutase (SOD)-like activity, scavenging activity against hydrogen peroxide and the polyphenol content of the garlic extract were increased 13-folds, more than 10-folds, and 7-folds, respectively, as compared with those of the control garlic extract. The results indicate that relatively short-term spontaneous fermentation potentiates anti-oxidative properties of garlic in fresh form, which is, at least in part, attributable to the increased level of polyphenols. Since superoxide is the primary upstream radical of the chain reaction with reactive oxygen species and hydrogen peroxide is generated from the scavenging reaction by SOD, the fermented- garlic is suggested to possess desirable anti-oxidative properties.     

Preparation and Characterization of Antioxidant Nanoparticles Composed of Chitosan and Fucoidan for Antibiotics Delivery

In this study, we developed novel chitosan/fucoidan nanoparticles (CS/F NPs) using a simple polyelectrolyte self-assembly method and evaluated their potential to be antioxidant carriers. As the CS/F weight ratio was 5/1, the CS/F NPs were spherical and exhibited diameters of approximately 230–250 nm, as demonstrated by TEM. These CS/F NPs maintained compactness and stability for 25 day in phosphate-buffered saline (pH 6.0–7.4). The CS/F NPs exhibited highly potent antioxidant effects by scavenging 1,1-diphenyl-2-picrylhydrazyl (DPPH), reducing the concentration of intracellular reactive oxygen species (ROS) and superoxide anion (O₂⁻) in stimulated macrophages. The DPPH scavenging effect of CS/F NPs primarily derives from fucoidan. Furthermore, these CS/F NPs activated no host immune cells into inflammation-mediated cytotoxic conditions induced by IL-6 production and NO generation. The MTT cell viability assay revealed an absence of toxicity in A549 cells after exposure to the formulations containing 0.375 mg NPs/mL to 3 mg NPs/mL. Gentamicin (GM), an antibiotic, was used as a model drug for an in vitro releasing test. The CS/F NPs controlled the release of GM for up to 72 h, with 99% of release. The antioxidant CS/F NPs prepared in this study could thus be effective in delivering antibiotics to the lungs, particularly for airway inflammatory diseases.     

Increased Level of Tetrahydro-β-Carboline Derivatives in Short-Term Fermented Garlic

In our previous study [1], we found that relatively short-term spontaneous fermentation (40 days at 60–70°C, 85–95% relative humidity) potentiates anti-oxidative properties of garlic, in which scavenging activity against hydrogen peroxide was included. Since tetrahydro-β-carboline derivatives (THβCs) that possess hydrogen peroxide scavenging activity have recently been identified in aged garlic extract, THβCs were quantitatively analyzed with liquid chromatography-mass spectrometry (LC-MS). (1R, 3S)-1-Methyl-1,2,3,4-tetrahydro-β-carboline-3-carboxylic acid (MTCC) and (1S, 3S)-MTCC were found in the fermented garlic extract whereas only trace levels of MTCCs were detected in the row garlic extract. Therefore, it is suggested that relatively short-term fermentation potentiates scavenging activity of garlic against hydrogen peroxide by forming THβCs, especially MTCCs.     

Antioxidant and Antiproliferative Activity of <Emphasis Type="Italic">Diospyros lotus</Emphasis> L. Extract and Isolated Compounds

The object of the study was to determine the chemical composition of Diospyros lotus L. extract and their antioxidant and antiproliferative properties. Eight compounds were isolated from D. lotus and identified as gallic acid, methylgallate, ellagic acid, kaempferol, quercetin, myricetin, myricetin 3-O-β-glucuronide, and myricetin-3-O-α-rhamnoside. D. lotus extract tested in different in vitro systems (DPPH, ABTS, FRAP, and Fe²⁺ chelating activity assay) showed significant antioxidant activity. The potential antiproliferative properties of D. lotus extract and isolated compounds against nine human cancer cell lines such as COR-L23, CaCo-2, C32, ACHN, A375, A549, Huh-7D12, MCF-7, and LNCaP were investigated in vitro by SRB assay. D. lotus extract demonstrated the highest inhibitory activity against COR-L23 with an IC₅₀ value of 12.2 μg/ml. Among identified hydrolysable tannins, ellagic acid evidenced strong antiproliferative activity against both C32 and A375 cells with IC₅₀ values of 0.8 and 4.1 μg/ml, respectively. Interesting results were observed, also, with gallic acid that showed the highest cytotoxic activity against CaCo-2 (IC₅₀ 2.6 μg/ml). Overall, the results of this study suggest that D. lotus displays a good antioxidant activity and has antiproliferative effects. Both activities are related to identified phenolic compounds.     

Studies on Indian Green Leafy Vegetables for Their Antioxidant Activity

To identify the potential of green leafy vegetables (GLV) as antioxidants, methanolic extracts of Amaranthus sp., Centella asiatica, Murraya koenigii and Trigonella foenum graecum were studied for their antioxidant activity in different systems at multiple concentrations. Total antioxidant activity assessed by phosphomolybdenum method, free radical scavenging activity by 1,1-diphenly-2-picryl hydrazyl (DPPH), reducing power and ferrous ion chelating activity were determined. The GLV were analyzed for ascorbic acid, total and β-carotene and total polyphenol contents. The ascorbic acid, total carotene, β-carotene and total phenolic content (tannic acid equivalents) of the GLV ranged between 15.18–101.36, 34.78–64.51, 4.23–8.84 and 150.0–387.50 mg/100 g GLV, respectively. The extracts were found to have significantly different levels of antioxidant activities in the systems tested. The total antioxidant activity was highest in Murraya koenigii (2,691.78 μmol of ascorbic acid/g sample) and least in Centella asiatica (623.78 μmol of ascorbic acid/g sample). The extract concentration causing 50% inhibition of DPPH (IC₅₀) was determined (M. koenigii < C.asiatica < Amaranthus sp. < T. graecum). The maximum DPPH scavenging activity and reducing power was exhibited by Murraya koenigii. Multiple regression analysis showed that the relationship of total antioxidant activity, free radical scavenging activity, and reducing power with polyphenol and total and β-carotene was highly significant. Paper awarded the Young Scientist Award in Experimental Nutrition at the Young Scientist Award Session of the 39th Annual Meet of the Nutrition Society of India, Hyderabad, India on 15–16th Dec 2007.     

Antioxidant Activities of Hydrolysates of Arca Subcrenata Prepared with Three Proteases

In order to get products with antioxidant activity from Arca subcrenata Lischke, the optimal hydrolase and hydrolysis conditions were investigated in the paper. Three proteases (neutrase, alcalase and papain) were applied to hydrolyze the homogenate of A. subcrenata. An orthogonal design was used to optimize hydrolysis conditions, and the pH-stat methods was used to determine the degree of hydrolysis. Viewed from the angle of reducing power, such as scavenging activities against α,α-diphenyl-β-picrylhydrazyl (DPPH) radical and hydrogen peroxide, the antioxidant activities of the alcalase hydrolysate (AH) were superior to neutrase hydrolysate (NH) and papain hydrolysate (PH), and its EC₅₀ values in DPPH radical and hydrogen peroxide scavenging effect were 6.23 mg/ml and 19.09 mg/ml, respectively. Moreover, compared with products hydrolyzed by neutrase and papain, the molecular mass of AH was lower and its content of amino acid of peptides was higher. Therefore, alcalase was selected as the optimal enzyme to produce active ingredients since its hydrolysate exhibited the best antioxidant activity among them and possessed large amount of potential active peptides.     

Polyphenolics Profile, Antioxidant and Radical Scavenging Activity of Leaves and Stem of Raphanus sativus L.

Aerial parts (leaves and stem) of Raphanus sativus, which are usually discarded were found to possess potent antioxidant and radical scavenging activity, as measured by standard antioxidant assays. Methanolic and acetone extracts of R. sativus leaves had total polyphenolic content of 86.16 and 78.77 mg/g dry extract, which were comparable to the traditional rich sources such as green tea and black tea. HPLC identification of polyphenolics indicated the presence of catechin, protocatechuic acid, syringic acid, vanillic acid, ferulic acid, sinapic acid, o-coumaric acid, myricetin, and quercetin in leaves and stem. Among the different extraction solvents, methanolic extract of leaves and stem showed potent reductive capacity, significantly inhibited linoleic acid peroxidation and displayed metal chelating activity. Further, they scavenged free radicals effectively with IC₅₀ (half maximal inhibitory concentration) of 31 and 42 μg/ml for DPPH radical, 23 and 52 μg/ml for superoxide radical, 67 and 197 μg/ml for hydrogen peroxide, and 56 and 62 μg/ml for nitric oxide, respectively. Leaves showed most potent antioxidant and radical scavenging activity as compared to stem, which may be accounted for the high polyphenolic content. Leaves and stem of R. sativus, often under-utilized part of this vegetable, thus possessed considerable amount of polyphenolics. Hence, it should be regarded as a potential source of natural antioxidants and could be effectively employed as an ingredient in health or in functional food.     

Valorization of the whole grains of Triticum aestivum L. and Triticum vulgare L. through the investigation of their biochemical composition and in vitro antioxidant,anti-inflammatory,anticancer and anticalpain activities

Two locally grown wheat species named Triticum aestivum L. and Triticum vulgare L. were studied for their phytochemical contents and their biological activities. T. vulgare presented the highest amounts of total phenolic compounds and ascorbic acids while T. aestivum was found to be rich in flavonoids, flavonols, proanthocynidins and ortho-diphenols. Eleven carotenoids were identified in T. vulgare where the most dominant compounds belongs to α-carotene and its derivatives while T. aestivum presented seven carotenoids. This later presented the highest DPPH radical scavenging activity and exhibited a strong reducing power in FRAP, phosphomolybdenum, hydrogen peroxide and reducing power assays. T. vulgare extract was found to be effective in metal chelating power and in scavenging nitric oxide radical. No significant differences in scavenging ABTS and hydroxyl radicals were noted between the two wheat species. T. aestivum inhibited xanthine oxidase and ROS production and showed the best cytotoxic effect while T. vulgare extract exhibited anti-calpain activity.     

Aged Garlic Extract Scavenges Superoxide Radicals

There is increasing evidence to suggest that many degenerative or pathological processes, such as aging, cancer, and coronary heart disease, are related to reactive oxygen species and radical-mediated reactions. We examined the effectiveness of aged garlic extract (AGE), a garlic preparation rich in water-soluble cysteinyl moieties, and its component for scavenging of superoxide by using the hypoxanthine-xanthine oxidase and human neutrophils. In the hypoxanthine-xanthine oxidase system, electron spin resonance showed that aged garlic extract scavenged superoxide radicals in a dose-dependent manner up to 54%. The EC₅₀ value of aged garlic extract for the superoxide radical scavenging effect was 0.80 mg/ml. N-α-(1-deoxy-D-fructos-1-yl)-L-arginine (25.9%) and (1S, 3S)-1-methyl-1,2,3,4-tetrahydro-β-carboline-1,3-dicarboxylic acid (20.8%), water-soluble moieties of AGE, also exerted superoxide scavenging effects. Phorbol 12-myristate 13-acetate-activated human neutrophils produced superoxide radical of 56.6 ± 9.27 nmol/min/10⁷ cells. Aged garlic extract (3 mg/ml) significantly inhibited superoxide production in comparison to the control. These data suggest that aged garlic extract may be useful for preventing diseases associated with reactive oxygen species.     

吲哚丁酸的体外抗氧化活性研究

研究吲哚丁酸的体外抗氧化活性,包括DPPH自由基、羟基自由基和过氧化氢的清除能力,以及金属螯合活性和还原力。结果表明,吲哚丁酸具有显著的抗氧化活性,且与吲哚乙酸相当,其对DPPH自由基和羟自由基清除作用、螯合力、还原能力均高于萘乙酸和2,4-二氯苯氧乙酸,而对Fe2+的络合能力与没食子酸无显著差异。     

Applicability of lignins from different sources as antioxidants based on the protective effects on lipid peroxidation induced by oxygen radicals

Increasing evidence suggest that excessive concentrations of reactive oxygen species in the human body are involved in a number of pathological events and that oxidative damage to cell components may play an important pathophysiological role in many types of human diseases. Several studies have shown the possible benefits of antioxidants from plant sources in altering, reversing or forestalling the negative effects of oxidative stress.The study was designed to evaluate whether lignins prevent H₂O₂-induced lipid peroxidation. The protective effect of industrial lignins obtained from different sources against hydrogen peroxide induced oxidation was investigated in normal human red blood cells (RBCs). RBCs, preincubated with different amounts of lignins and challenged with hydrogen peroxide were analyzed for lipid peroxidation. The high molecular weight can be considered as one of the main factors decreasing the radical scavenging activity. When comparing different lignins the inhibitory effects against lipid peroxidation were notably related to their molecular weights. In this sense bagasse with the lowest Mn was the greatest antioxidant and lignosulfonate with the highest Mn was the lowest antioxidant lignin.These results open the possibility of new applications of lignins. The potential health benefits and industrial applications of lignins make them a promising research resource.     

伏安极谱法测定槟榔提取物对过氧化氢自由基的清除活性

采用微分脉冲伏安法及循环伏安法测定不同溶剂(水、氯仿、正已烷、丙酮、乙酸乙酯)槟榔提取物对过氧化氢(H2O2)自由基的清除能力。结果显示,用Vc和咖啡酸作对照,不同溶剂槟榔提取物均有清除过氧化氢自由基的能力,其中乙酸乙酯提取物的最高,水提取物的次之,氯仿、正已烷和丙酮提取物的清除能力略低。表明伏安极谱法可用作评价天然植物提取物清除H2O2活性大小的一种快速检测的分析方法。     

Identifying Potential Antioxidant Properties from the Viscera of Sea Snails (Turbo cornutus)

Turbo cornutus, the horned turban sea snail, is found along the intertidal and basaltic shorelines of Jeju Island, Korea. T. cornutus feeds on seaweeds (e.g., Undaria sp., and Ecklonia sp.) composed of diverse antioxidants. This study identified potential antioxidant properties from T. cornutus viscera tissues. Diverse extracts were evaluated for their hydrogen peroxide (H₂O₂) scavenging activities. T. cornutus viscera protamex-assisted extracts (TVP) were purified by gel filtration chromatography (GFC), and potential antioxidant properties were analyzed for their amino acid sequences and its peroxidase inhibition effects by in silico molecular docking and in vitro analysis. According to the results, T. cornutus viscera tissues are composed of many protein contents with each over 50%. Among the extracts, TVP possessed the highest H₂O₂ scavenging activity. In addition, TVP-GFC-3 significantly decreased intracellular reactive oxygen species (ROS) levels and increased cell viability in H₂O₂-treated HepG2 cells without cytotoxicity. TVP-GFC-3 comprises nine low molecular bioactive peptides (ELR, VGPQ, TDY, ALPHA, PAH, VDY, WSDK, VFSP, and FAPQY). Notably, the peptides dock to the active site of the myeloperoxidase (MPO), especially TDY and FAPQY showed the MPO inhibition effects with IC₅₀ values of 646.0 ± 45.0 µM and 57.1 ± 17.7 µM, respectively. Altogether, our findings demonstrated that T. cornutus viscera have potential antioxidant properties that can be used as high value-added ingredients.