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.     

Potential mechanism of kaempferol against Cu2+-induced oxidative stress through chelating activity and regulation of nuclear factorerythroid-2-related factor 2 signaling

Metal ions play important roles in various biological processes of living systems. However, the generation of reactive oxygen species (ROS) is also closely linked with the participation of redox-active metal ions such as copper, iron, chromium, and cobalt ions. Excessive production of ROS by redox-active metal ions can cause oxidative stress and further oxidative stress-related diseases. This study shows the results of the antioxidant activity of kaempferol in both ORAC_OH· and Cu²⁺-treated HepG2 cells. Preventive mechanism of kaempferol in Cu²⁺-treated HepG2 cells is also elucidated. These results suggest that both cellular Cu²⁺-chelating activity and expression of phase II detoxifying enzymes such as HO-1 and GSTA2 through activating Nrf2 are required for cellular antioxidant activity of kaempferol in Cu²⁺-treated HepG2 cells. Our findings provide the scientific evidence for the development of Nrf2 targeting dietary antioxidant to prevent oxidative stressrelated conditions.     

蜜柚果肉膳食多酚的结构鉴定及抗氧化机理

蜜柚果肉富含膳食多酚，极具保健价值，但其理论研究不足导致深加工产业受限。本实验以梅州蜜柚为原料，对其果肉膳食多酚进行粗提和表征，并研究其对H2O2诱导的HepG2细胞氧化应激的保护作用及其潜在作用机理。结果显示，在柚果肉膳食多酚粗提物中共鉴定出7 种可精确定量多酚单体：柚皮苷、芸香柚皮苷、野漆树苷、香草酸、甲基橙皮苷、新西兰牡荆苷和阿魏酸，其中柚皮苷含量为50.54 mg/g，显著高于其他6 种多酚含量。新西兰牡荆苷和野漆树苷为首次在蜜柚果肉中发现。质量浓度为20 μg/ mL和100 μg/mL的膳食多酚粗提物对H2O2诱导的HepG2肝癌细胞氧化损伤均具有保护作用，且能降低该细胞中乳酸脱氢酶的释放，降低活性氧的积累，促进核因子E2相关因子2（nuclear factor erythroid 2-related factor 2，Nrf2）从细胞质到细胞核的转运，并上调其靶基因GSTA2、HO-1、GCLC、NQO1的表达。综上，梅州蜜柚果肉膳食多酚可以作为预防氧化应激相关疾病潜在的抗氧化剂。     

Comparison of constituents, antioxidant potency, and acetylcholinesterase inhibition in Lentinus edodes, Sparassis crispa, and Mycoleptodonoides aitchisonii

This study evaluates three edible mushrooms: Lentinus edodes, Sparassis crispa, and Mycoleptodonoides aitchisonii, in terms of acetylcholinesterase (AChE) inhibition, antioxidant potency and constituents free amino acids and mineral. The concentration of essential amino acids was found to be 34.10 mg/g in M. aitchisonii, 26.25 mg/g in S. crispa, 25.99 mg/g in L. edodes. S. crispa displayed the highest DPPH scavenging activity and phenolic contents. The best results for AChE inhibition were obtained from M. aitchisonii. These results suggest that M. aitchisonii has high potential for cognitive improvement by AChE inhibition and antioxidant potency.     

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.     

Comparative antioxidant, antiproliferative and phase II enzyme inducing potential of sorghum (Sorghum bicolor) varieties

Epidemiological evidence has linked consumption of sorghum with reduced incidences of gastrointestinal cancer, especially cancer of esophagus. No information is available on how sorghum may effect the chemoprotective properties. We investigated in vitro potential of eight sorghum varieties to induce phase II detoxifying enzymes using the NAD(P)H:quinone oxidoreductase (NQO) enzyme assay, and also inhibit proliferation of esophageal, OE33, and colon, HT-29, carcinoma cells using the PicoGreen and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays; these properties were compared to phenolic profile and antioxidant activity of the sorghum. Black sorghum extract high in 3-deoxyanthocyanins was the most potent NQO inducer, doubling NQO activity at 5.0 μg/mL and maximally inducing the enzyme activity by 3.0 times. White sorghum was a moderately strong inducer, maximally increasing NQO activity by 80%; tannin-containing sorghums were non-inducers. On the other hand, the tannin-containing sorghum extracts had strongest antiproliferative activity against both OE33 and HT-29 cells (IC₅₀, 38-105 μg/mL); the white sorghum extract was the least potent (IC₅₀, 389->800 μg/mL). Antiproliferative activity correlated with antioxidant activity whereas NQO-inducer capacity did not. Sorghum extracts have strong chemoprotective potential which is partially independent of their antioxidant properties. They may thus be valuable health-promoting ingredients in whole-grain based products.     

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.     

槲皮素通过Nrf2通路对糖尿病大鼠胰腺氧化损伤的拮抗作用机制

目的：探讨槲皮素是否通过核因子E2相关因子2（nuclear factor erythroid 2-related factor 2, Nrf2）通路拮抗糖尿病大鼠胰腺氧化损伤。方法：选取SPF级SD大鼠48 只,随机选取10 只大鼠作为正常对照组,其余38 只大鼠饲喂高脂饲料联合注射30 mg/kg mb链脲佐菌素建立2型糖尿病（type 2 diabetes mellitus,T2DM）模型。以大鼠随机血糖浓度不低于16.7 mmol/L作为造模成功判断标准,按血糖浓度分层将大鼠随机分为T2DM模型组、低剂量槲皮素（L-QU）、高剂量槲皮素（H-QU）干预组,每组10 只,连续灌胃12 周。测定大鼠胰腺组织匀浆液的氧化损伤和抗氧化指标,运用Western Blot、实时定量聚合酶链式反应测定Nrf2、血红素氧合酶1（heme oxygenase 1,HO-1）、谷胱甘肽硫转移酶（glutathione S-transferase,GST）、NADP(H):醌氧化还原酶1（NADP(H):quinone oxido-reductase 1,NQO1）蛋白及其mRNA表达量。结果：与T2DM模型组相比,低、高剂量槲皮素干预显著降低了大鼠血糖浓度和胰腺丙二醛含量（P＜0.05）,减轻胰岛素抵抗,显著提高超氧化物歧化酶等抗氧化酶活力（P＜0.05）,Nrf2以及下游抗氧化酶HO-1、NQO1的蛋白含量和mRNA表达量均显著上升（P＜0.05）。结论：槲皮素可能是通过激活胰腺组织Nrf2信号通路,上调下游抗氧化酶表达,进而减轻T2DM大鼠胰腺氧化损伤,改善胰岛素抵抗,调节血糖水平,从而发挥防治糖尿病的作用。     

Collagen peptides from Acaudina molpadioides prevent CCl4-induced liver injury via Keap1/Nrf2-ARE,PI3K/AKT,and MAPKs pathways

Collagen peptide from Acaudina molpadioides (AMP) showed antioxidative activity in H₂O₂-induced RAW264.7 cells in our pervious study. In this study, it was observed that AMP could effectively improve the morphology and function of liver in CCl₄-induced mice. After 200 mg/kg AMP treatment, the content of MDA in liver decreased by 62.3%, and the level of antioxidant enzymes (SOD, GSH-Px, and CAT) increased by more than 65%. Western blot results disclosed that AMP (200 mg/kg) upregulated the Nrf2 level by 73.8% and downregulated Keap1 by 41.0% in CCl₄-induced mice liver. The levels of p-ERK, p-JNK, and p-p38 in 200 mg/kg AMP treatment groups decreased by 57.3%, 40.9%, and 40.6%, but the levels of p-PI3K and p-AKT increased by 162.6% and 60.3%, respectively. Furthermore, the trends of Nrf2, Keap1, p-ERK, p-JNK, p-p38, p-PI3K, and p-AKT levels in H₂O₂-induced RAW264.7 cells after AMP treatment were similar to the results in CCl₄-induced mice liver. These findings provided evidence that AMP exerted antioxidant activity via Keap1/Nrf2-ARE, PI3K/AKT, and MAPKs pathways in vivo and in vitro. Therefore, the collagen peptide from A. molpadioides might represent a novel functional food to prevent acute liver injury via attenuation of oxidative stress.     

Ferulic acid released by treatment with Aspergillus oryzae contributes to the cellular antioxidant capacity of wheat germ extract

An extract of fermented wheat germ (EFWG) was prepared by treatment with Aspergillus oryzae and in vitro antioxidant and cellular antioxidant capacities were measured. The induction of phase II detoxifying and antioxidant enzymes in exerting a cellular antioxidant capacity was determined. Treatment of wheat germ with A. oryzae significantly (p<0.001) increased the ferulic acid content, compared to controls, among 4 phenolic acids analyzed. The peroxyl radical-scavenging and reducing capacities and the cellular antioxidant capacity of EFWG were more potent than for extracts of wheat grain (EWG). The cellular antioxidant capacity of EFWG against AAPH and H₂O₂-induced oxidative stress was stronger than for EWG due to enhanced induction of the phase II enzymes HO-1, GST, and NQO-1. Potent in vitro and cellular antioxidant capacities of EFWG may result from an increased ferulic acid content due to treatment with A. oryzae.     

沙棘花青素提取物对双氧水诱导的H1299细胞损伤的保护作用及Nrf2/HO-1通路的影响

目的：研究沙棘花青素提取物（The anthocyanidin extract of Hippophae rhamnoides L.,HRAE）对双氧水诱导H1299细胞氧化损伤的保护作用及其机制。方法：利用双氧水诱导H1299细胞氧化损伤模型,采用MTT法检测HRAE对细胞活力的影响,采用荧光探针DCFH-DA检测细胞内活性氧（Reactive oxygen species,ROS）的含量;采用试剂盒分别测定超氧化物歧化酶（Superoxide dismutase,SOD）、过氧化氢酶（Catalase,CAT）、谷胱甘肽过氧化物酶（Glutathion peroxidase,GSH-Px）及丙二醛（Malondialdehyde,MDA）的含量;采用Western Blot法检测血红素氧合酶-1(Heme oxygenase-1,HO-1)、醌氧化还原酶-1(NAD(P)H quinine oxidoreductase 1,NQO1)、Kelch样环氧氯丙烷相关蛋白1(Kelch like ECH-associated protein 1,Keap1)和核转录因子E2相关因子（Nu...     

芦荟多糖对D-半乳糖致HepG2细胞氧化损伤的保护作用

目的:探讨芦荟多糖对HepG2细胞氧化损伤的保护作用,分析其体外抗氧化能力。方法:以HepG2细胞为研究对象,建立D-半乳糖诱导细胞氧化损伤模型,以不同浓度芦荟多糖进行预保护处理。采用细胞增殖与毒性检测试剂盒测定HepG2细胞存活率,生物化学法检测细胞超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶活力,酶联免疫吸附法测定细胞总抗氧化能力及丙二醛（malondialdehyde,MDA）含量,实时荧光定量PCR检测细胞Keap1、Nrf2、NQO1和HO-1基因表达水平。结果:与D-半乳糖模型组比较,25、50、100 μg/mL的芦荟多糖预保护处理能不同程度地提高HepG2细胞存活率;芦荟多糖能显著增强细胞中抗氧化酶的活性,降低细胞MDA的生成量（P<0.05）,且作用效果与芦荟多糖浓度成正比;细胞中Keap1基因表达显著降低,Nrf2、NQO1、HO-1 mRNA表达显著提高（P<0.05）。结论:芦荟多糖可以减轻HepG2细胞的氧化应激损伤,激活Nrf2表达并抑制其泛素化降解,提高下游NQO1、HO-1的转录水平,增强细胞抗氧化酶系活性并调控细胞氧化还原系统,以达到减轻细胞氧化损伤程度、提高细胞抗氧化应激损伤的效果。     

Evaluation of the anti-inflammatory efficacy of Lotus corniculatus

The anti-inflammatory effects of the crude extract (CE) of Lotus corniculatus var. São Gabriel and its derived hexane (HEX), ethyl acetate (AcOEt), n-butanol (BuOH) and aqueous (Aq) fractions and isolated compounds kaempferitrin, oleanolic acid and β-sitosterol, in a mouse model of pleurisy induced by carrageenan were investigated. Swiss mice were used in the in vivo experiments. The crude extract of L. corniculatus and its derived fractions, and also its isolated compounds, inhibited leukocytes, exudation, and myeloperoxidase (MPO) and adenosine-deaminase (ADA) activities, as well as nitrite/nitrate concentration and interleukin-1 beta (IL-1β) level (p < 0.05). L. corniculatus showed important anti-inflammatory activity by inhibition not only of leukocytes and/or exudation, but also of pro-inflammatory enzymes and mediators such as MPO, ADA, and IL-1β. The constituent’s kaempferitrin, oleanolic acid and β-sitosterol may well account for it.     

Compounds from Angelica keiskei with NQO1 induction, DPPH scavenging and α-glucosidase inhibitory activities

A LC-MS method, which GSH was used as substrate, was employed to reveal the compounds with NQO1 induction activity from Angelica keiskei. Some compounds, proposed as isobavachalcone, xanthoangelol and 4-hydroxyderricin, have NQO1 induction activity. To make the actual structures and bioactivities of these compounds clear, 23 compounds, including above mentioned compounds and two new compounds 4-hydroxy-3,5,5-trimethyl-4-(1,2,3-trihydroxybutyl)cyclohex-2-enone (18) and (Z)-2-(3-hydroxypent-1-ynyl)-3-(non-1-enyl)oxiran-2-ol (23), were isolated from the 95% ethanol extract of A. keiskei. The bioassay results suggested the compounds had notable NQO1 induction activity. The radical scavenging and α-glucosidase inhibition activities of the isolated compounds were also tested. Compounds (E)-1-(2,4-dihydroxy-3-(3-methylbut-2-enyl)phenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one (1), (E)-1-(3-((E)-3,7-dimethylocta-2,6-dienyl)-2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one (2), 1-(3-hydroxy-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[α]phenanthren-17-yl)ethanone (17), 4-hydroxy-3,5,5-trimethyl-4-(1,2,3-trihydroxybutyl)cyclohex-2-enone (18) could scavenge DPPH radical by more than 20%. Compounds (E)-1-(2,4-dihydroxy-3-(3-methylbut-2-enyl)phenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one (1), (E)-1-(3-((E)-3,7-dimethylocta-2,6-dienyl)-2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one (2), (E)-1-(2-hydroxy-4-methoxy-3-(3-methylbut-2-enyl)phenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one (3), 7-hydroxy-6-(3-methylbut-2-enyl)-2H-chromen-2-one (5), 10-hydroxy-8,8-dimethyl-2-oxo-9,10-dihydropyrano[6,5-H]chromen-9-yl)-3-methylbut-2-enoate (9), 2-(4-hydroxyphenyl)-7-methoxy-8-(3-methylbut-2-enyl)-2,3-dihydrochromen-4-one (16), (10S,15R,Z)-10,15-dihydroxyheptadeca-8,16-dien-11,13-diynyl acetate (20), (3R,8S,Z)-heptadeca-1,9-dien-4,6-diyne-3,8-diol (21) exhibited excellent α-glucosidase inhibition activity.     

Ultraviolet irradiation: The generator of Vitamin D2 in edible mushrooms

Fresh Shiitake mushrooms (Lentinula edodes), Oyster mushrooms (Pleurotus ostreatus), Button mushrooms (Agaricus bisporus), and Abalone mushrooms (Pleurotus cystidus) were irradiated with Ultraviolet-A (UV-A; wavelength 315–400 nm), Ultraviolet-B (UV-B; wavelength 290–315 nm), and Ultraviolet-C (UV-C; wavelength 190–290 nm). Irradiation of each side of the mushrooms for 1 h, was found to be the optimum period of irradiation in this conversion. The conversions of ergosterol to vitamin D₂ under UV-A, UV-B, and UV-C were shown to be significantly different (p < 0.01). The highest vitamin D₂ content (184 ± 5.71 μg/g DM) was observed in Oyster mushrooms irradiated with UV-B at 35 °C and around 80% moisture. On the other hand, under the same conditions of irradiation, the lowest vitamin D₂ content (22.9 ± 2.68 μg/g DM) was observed in Button mushrooms.     

Isolation,characterization, and neuroprotective activities of sesquiterpenes from Petasites japonicus

Neuroprotective reagents to protect the nerve cells against oxidative stress and other damages are potentially effective for the medical treatment of Parkinson’s disease. Petasites japonicus, a wild vegetable, belongs to the family Compositae and its extract has shown the neuroprotective effects. A further phytochemical investigation of P. japonicus for neuroprotective substances led to the isolation of eight new (1–8) and two known (9 and 10) sesquiterpenes. Their structures were elucidated on the basis of extensive 1D and 2D NMR (HMQC, HMBC, ¹H–¹H COSY, and NOESY) spectroscopic data analyses, and the structure of 1 was confirmed by X-ray crystallography. The neuroprotective activities of these sesquiterpenes were evaluated against cobalt chloride (CoCl₂)-induced neuronal cell death in human dopaminergic SH-SY5Y cells. Five compounds showed a neuroprotective activity.