耐辐射球菌清除活性氧自由基及对DNA的保护作用

用化学发光法分析了耐辐射球菌 (D .radiodurans)的两个菌株 (R1、KD830 1 )和大肠杆菌对超氧阴离子自由基、羟基自由基和过氧化氢的清除能力。通过羟基自由基诱导的DNA氧化损伤化学发光模型考察了耐辐射球菌细胞提取物对自由基氧化导致DNA氧化损伤的保护作用。结果表明 ,耐辐射球菌提取物能显著清除O-2·、H2 O2 和·OH ,清除能力明显高于大肠杆菌 ,并能有效地防止自由基所致DNA氧化损伤 ,表现出超强的抗氧化和耐辐射能力 ,这归因于耐辐射球菌中具有较高的抗氧化酶活性     

菜籽多酚和Vc在化学模拟体系中清除超氧阴离子和羟自由基的能力

研究菜籽多酚、Vc、菜籽多酚与Vc混合物在化学模拟体系中清除超氧阴离子自由基OH.O2.与羟自由基.OH自由基的能力。用邻苯三酚自氧化法测定抗氧化剂清除OH.O2.的能力,用铁氧化邻二氮菲-Fe2+法测定抗氧化剂清除.OH的能力。结果表明:菜籽多酚、Vc、菜籽多酚与Vc的混合物清除OH.O2.、.OH都不呈剂量-效应关系。菜籽多酚在0.25mg/ml浓度时对OH.O2.与.OH的清除率均最高,分别达51.40%和84.82%。Vc在0.80mg/ml时对OH.O2.的清除率最高,达80.39%;在0.40mg/ml时对.OH的清除率最高,达91.35%。当总浓度为0.25mg/ml时,菜籽多酚与Vc的混合物没有协同增加清除OH.O2.的作用;菜籽多酚与Vc按7∶3、8∶2两种比例混合,在清除.OH时有协同增效作用,协同增效的最佳比例为7∶3,最大清除率达78.78%。     

重金属胁迫对水稻不同品种超氧化物歧化酶的影响

用不同浓度的Cd2 + 、Cu2 + 和Hg2 + 处理花期的扬稻 6号 (籼稻 )、武育粳 8号 (粳稻 )和协优 81 8(杂交稻 ) ,测定各器官SOD活性 ,并进行SOD同功酶的电泳分析。结果表明 ,较高浓度重金属显著抑制各品种根系SOD活性 ,也抑制叶片SOD活性 ,但抑制程度因重金属种类和品种类型而异。各浓度均抑制武育粳穗SOD活性 ,但诱导扬稻 6号穗SOD活性增高。协优 81 8穗SOD活性受Cd2 + 抑制 ,但却受高浓度Cu2 + 和Hg2 + 诱导。≥ 2 0ppmCd2 + 、Cu2 + 和Hg2 + 均抑制水稻根系SOD同功酶表达 ,但诱导扬稻 6号和协优 81 8穗SOD同功酶表达。Mn SOD和Cu Zn SOD同功酶对重金属胁迫的敏感性不同 ,重金属对水稻根系Mn SOD的抑制程度明显高于Cu Zn SOD。     

DEHP对土壤蚯蚓氧化胁迫及DNA损伤的研究

土壤环境中的酞酸酯污染日益严重,为了探讨和分析典型酞酸酯邻苯二甲酸二(2-乙基己基)酯(Di(2-ethylhexyl)phthalate,DEHP)对土壤动物的生态毒理效应,以赤子爱胜蚓为指示生物,暴露于DEHP浓度为CK、0.1、1、10、50 mg kg~(-1)人工土壤中,并于染毒后的7、14、21、28d取样测定。通过蚯蚓体内的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物(POD)、谷胱甘肽转移酶(GST)等指标反映DEHP对蚯蚓的氧化激活程度,通过活性氧自由基(ROS)的含量反映DEHP对蚯蚓的细胞毒性,通过丙二醛(MDA)含量和Olive尾矩反映DEHP对蚯蚓的遗传毒性,从多个角度评价DEHP对蚯蚓的生态毒理效应。结果表明:(1)在DEHP的刺激下,各浓度组蚯蚓的SOD、CAT、POD、GST活性均呈激活状态,表明DEHP对蚯蚓的抗氧化酶有诱导作用;(2)DEHP影响蚯蚓的ROS含量,各染毒处理组与对照组相比均升高且差异性明显,表现出明显的剂量—效应关系;(3)对比各染毒处理组之间的数据,DEHP对蚯蚓MDA含量的影响无明显规律;(4)DEHP可使蚯蚓GST呈激活状态,表明中高浓度的DEHP对蚯蚓GST具有诱导作用;(5)DEHP能够引起蚯蚓体腔细胞DNA的损伤,且随着浓度的增加,Olive尾矩值随之增加,说明DNA损伤程度与DEHP浓度之间具有剂量-效应关系。从实验结果可以看出,DEHP可以对蚯蚓机体和DNA造成一定程度的损伤,表现出较强的生态毒理效应。     

赭曲霉毒素A引起HepG2细胞损伤及DNA甲基化水平降低

赭曲霉毒素A (ochratoxinA,OTA)是一种污染食品的真菌毒素,对许多动物具有肾毒性,肝毒性,免疫毒性,致畸和致癌作用.本研究以人肝癌细胞HepG2为体外模型,研究OTA引起的细胞损伤及对DNA稳定性的影响.MTT结果显示,OTA能抑制HepG2细胞的生长,存在时间剂量——效应关系.OTA引起细胞上清中酸脱氢酶(LDH),谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性的升高,诱导细胞内活性氧(ROS)生成,降低超氧化物歧化酶(SOD)活力.彗星实验结果显示OTA引起DNA链断裂,形成拖尾现象.同时,本研究还首次研究了OTA对DNA甲基化的影响.HPLC/MS/MS结果显示,经OTA处理后的细胞,其DNA中5-甲基脱氧胞苷(5mdC)百分数显著低于对照组.这些结果表明OTA会抑制HepG2细胞生长,引起氧化损伤,破坏DNA的稳定性和完整性.本研究表明OTA引起的肝毒性可能与氧化损伤和DNA损伤有关.     

灵芝肽在卵磷脂体系中的抗氧化作用研究

卵磷脂是生物膜的重要构成成分,易受到自由基的攻击而氧化.用TBA法对灵芝肽在卵磷脂体系中的抗氧化活性进行了研究;同时采用脱氧核糖法研究了灵芝肽在非脂质体系中抑制OH的能力及用电位法测定了各种灵芝肽的氧化还原电位.结果表明在100 μmol/L抗坏血酸存在的条件下,灵芝肽能较好地抑制10 μmol/L Cu2+催化的卵磷脂体系的脂质氧化,且呈剂量效应;其中肽的抑制能力优于氨基酸.各种灵芝肽均有抑制羟基自由基的能力,其中以混合肽的抑制效果最佳,且混合肽的还原能力也是最强的.     

γ射线辐照对春兰根状茎生长及抗氧化酶活性的影响

将春兰(Cymbidium goeringii)的离体根状茎切成长各约1cm的顶段、中段和末段,用60Coγ射线进行辐照,剂量分别为0、5、10、20、30和50Gy。测定了不同辐照剂量下新生根状茎的生长,研究了辐照原初根状茎愈创木酚过氧化物酶(G-POD)、过氧化氢酶(CAT)和超氧化物歧化酶(SOD)等抗氧化酶活性以及过氧化物酶同工酶谱的影响。结果表明:辐照剂量为30和50 Gy处理的根状茎虽能存活,但未有新生根状茎的发生,生长停滞。辐照原初根状茎上CAT和SOD的活性随着辐照剂量的增加均有先升高后降低的趋势,而G-POD的活性则表现为先略降低后升高。综合新生根状茎的生长、抗氧化酶活性及过氧化物同工酶谱的结果,春兰诱变育种的适宜剂量应在20~30Gy间,且最合适的诱变部位是根状茎的顶段。     

电子束与γ辐射对大麦种胚DNA非预定合成的影响

羟基脲抑制与流式细胞仪检测证明 ,在大麦种子的吸涨初期 ,种胚DNA合成主要是DNA非预定合成 ,一定剂量下6 0 Co γ辐射能刺激增强种胚的DNA非预定合成。辐射刺激效应随修复时间延长呈下降趋势。 50～ 40 0Gy的电子束辐射对大麦种胚的DNA非预定合成也有刺激作用。在半致死剂量附近有一峰值 ,在致死剂量附近出现最小值。     

用~3H-TdR研究混合重金属对鲫鱼DNA合成的影响

应用3 H 胸腺嘧啶核苷 ( 3 H TdR)示踪法研究混合重金属对鲫鱼 (Carassiusaura tus)遗传毒性的影响。结果表明 ,最佳取样时间为注射3 H TdR后 6h。鱼体各组织对混合重金属的毒性反应呈现双向效应 ,即低浓度时 ,表现出损伤、修复作用 ;高浓度时 ,DNA合成受到抑制。鱼卵、鱼鳃对重金属最敏感 ,损伤浓度为 0 0 773mg L ;而大脑对混合重金属耐受性最强。DNA损伤修复作用和DNA合成作用都存在剂量效应 ,随着暴露时间的延长 ,毒性作用增大。鱼体各组织对重金属的毒性反应可由DNA损伤修复变成合成抑制。经过 72h暴露后 ,3 H TdR在肝脏、鱼鳃、鱼卵中掺入量约低于对照的 5 0 %。     

氮离子注入对仿栗种子当代生理生化性状的影响

研究了不同剂量的氮离子注入仿栗种子对种子内过氧化物酶、过氧化氢酶、超氧化物歧化酶活性以及丙二醛的含量、电解质外渗率的影响。结果表明:离子注入剂量在9×1016~12×1016N+/cm2下过氧化物酶、过氧化氢酶、超氧化物歧化酶的酶活性达到峰值,丙二醛含量和电解质外渗率最低,在一定程度上削弱种子脂质过氧化作用。结果说明一定剂量的氮离子注入能激活合成自由基清除酶的能力,在一定程度上免受自由基侵害。     

葛根素对小白菜镉损伤的保护作用研究

为探究葛根素(PUE)对小白菜镉(Cd)损伤的保护效果,采取外源添加不同葛根素处理,将小白菜植株随机分为7组,即对照组(CK)、PUE30组(葛根素30μmol·L^-1)、Cd4组(镉4 mg·L^-1)、Cd与PUE共处理组(4+15、4+30、4+45、4+60)。处理30 d后,观察小白菜植株生长状况、测定Cd富集量及抗胁迫生理活性物质,包括叶绿素、抗氧化酶(SOD、APX、CAT、POD)活性及过氧化氢(H₂O₂)、丙二醛(MDA)Vc、可溶性蛋白含量,并用实时荧光定量PCR(qRT-PCR)技术检测抗氧化酶基因(SOD、POD、APX及CAT)表达量。结果表明,与CK组相比,Cd4组小白菜根中Cd含量、H₂O₂和MDA含量显著升高,而地下部鲜重、地上部鲜重、抗氧化酶(SOD、POD和APX)活性和叶绿素含量整体显著降低(P<0.05)。与Cd4组相比,Cd+PUE共处理组小白菜氧化损伤程度减轻,H₂O₂     

Antioxidant effects of extracts from Cassia tora L. prepared under different degrees of roasting on the oxidative damage to biomolecules

The effects of water extracts from Cassia tora L. (WECT) treated with different degrees of roasting (unroasted and roasted at 150, 200, and 250 degrees C) on the oxidative damage to deoxyribose, DNA, and DNA base in vitro were investigated. It was found that WECT alone induced a slight strand breaking of DNA. In the presence of Fe(3+)/H(2)O(2), WECT accelerated the strand breaking of DNA at a concentration of 2 microg/mL; however, it decreased with increasing concentrations (>5 microg/mL) of WECT. WECT also accelerated the oxidation of deoxyribose induced by Fe(3+)-EDTA/H(2)O(2) at a concentration of 0.2 mg/mL but inhibited the oxidation of deoxyribose induced by Fe(3+)-EDTA/H(2)O(2)/ascorbic acid. Furthermore, WECT accelerated the oxidation of 2'-deoxyguanosine (2'-dG) to form 8-OH-2'-dG induced by Fe(3+)-EDTA/H(2)O(2). The prooxidant action of WECT on the oxidation of 2'-dG was in the order of unroasted > roasted at 150 degrees C > roasted at 200 degrees C > roasted at 250 degrees C. The decrease in the prooxidant activity of the roasted sample might be due to the reduction in its anthraquinone glycoside content or the formation of antioxidant Maillard reaction products after roasting. Thus, WECT exhibited either a prooxidant or an antioxidant property in the model system that was dependent on the activities of the reducing metal ions, scavenging hydroxyl radical, and chelating ferrous ion.     

大气O3浓度升高对麦田土壤重金属铜生物有效性和生理毒性的影响

在开放式空气O3浓度增加（FACE）平台下,采用盆栽试验,初步研究了O3浓度升高后麦田重金属Cu的生物有效性变化以及对各生长阶段小麦叶片生理毒性的影响。结果表明,在FACE条件下,小麦地上部对Cu的吸收相比于正常大气对应组有增加的现象;与正常大气对应组相比,FACE条件下土壤中有效态Cu的含量也有所增加;随着小麦的生长发育,FACE圈小麦叶片内的MDA含量总体呈上升趋势,O3升高铜污染组的小麦叶片内MDA的含量最高;与正常大气对照组相比,O3升高铜污染组的小麦叶片SOD酶和POD酶比较敏感,在分蘖期其活性受到诱导,但随着暴露时间的增加,抗氧化系统的各个酶的活性逐渐受到抑制。O3加剧了Cu对小麦的牛理胁迫．增加了Cu的牛物有效性。     

Antioxidant,prooxidant, and cytotoxic activities of solvent-fractionated dandelion (Taraxacum officinale) flower extracts in vitro

This study was conducted to investigate the chemical antioxidant and bioactive properties of the water (WF) and ethyl acetate fractions (EAF) derived from dandelion (Taraxacum officinale) flower extract (DFE). HPLC analysis showed the presence of both luteolin and luteolin 7-glucoside in the DFE, which contributed to noted in vitro antioxidant and Caco-2 cell cytotoxic activities. Both WF and EAF of DFE exhibited free radical scavenging activities in a stable 2,2-diphenyl-1-picrylhydrazyl radical model and reduced the breakage of supercoiled DNA strand induced by both non-site-specific and site-specific hydroxyl radical. Oxidation of structured phosphatidylcholine liposome induced by peroxyl radical was reduced in the presence of both EAF and WF. EAF had greater (p < 0.05) affinity to scavenge peroxyl radical than WF, as measured by the formation of conjugated diene. At low concentration, prooxidant activity of both fractions was observed in Cu(2+)-induced structured liposome and hLDL oxidation models, thus indicating that the reducing power of the DFE had resulted in generation of reactive cuprous ion. However, at high concentrations the EAF did not promote oxidation in the presence of Cu(2+), suggesting that the free radical scavenging activity of this fraction was sufficient to minimize the potential oxidative mechanism attributed to the metal ion reducing activity associated with prooxidant activity.     

Isolation and characterization of antioxidant protein fractions from melinjo (Gnetum gnemon) seeds

The protein from the seeds of melinjo ( Gnetum gnemon ) was purified using a precipitation method and ion exchange chromatographic techniques to identify the potent antioxidant and free radical scavenging activities. Two antioxidant protein fractions were isolated from G. gnemon seed with molecular weights of approximately 30 kDa (Gg-AOPI) and 12 kDa (Gg-AOPII) by SDS-PAGE. The N-terminal amino acid sequence of Gg-AOPII is Gly-Asn-Gly-Lys-Ala-Thr-Val-Ala-Ile-Leu-Val-Lys-Glu-Lys-Val-Glu-Tyr-Gly-Glu-Glu, and the result of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis showed that they were distinct from each other; no protein in database matching was found to both Gg-AOPI and Gg-AOPII. The antioxidant or free radical scavenging activities of Gg-AOPs were investigated by employing in vitro assay systems including the inhibition of linoleic acid autoxidation, scavenging effect on α,α-diphenyl-β-picrylhydrazyl free radical (DPPH), 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), reducing power, chelating abilities of metal ions Cu(2+) and Fe(2+), and protections against hydroxyl radical-mediated DNA damages. The result showed that two protein fractions exhibited significant (p < 0.05) antioxidant activities against free radicals such as DPPH, ABTS, and superoxide anion and showed activities similar to those of glutathione (G-SH) and BHT in a linoleic acid emulsion assay system. Moreover, Gg-AOPI and Gg-AOPII also exhibited notable reducing power and strong chelating effect on Fe(2+) and protected hydroxyl radical induced oxidative DNA damage. The data obtained by the in vitro systems obviously established the antioxidant potency of Gg-AOPs.     

Effect of lactoferrin on oxidative stability of corn oil emulsions and liposomes

Interest in using lactoferrin in foods for its antimicrobial activity inspired the present study of its antioxidant activity. Natural bovine lactoferrin inhibited oxidation in buffered corn oil emulsions and lecithin liposome systems at pH 6.6 and 50 degrees C. The antioxidant activity increased with lactoferrin concentration in both phosphate- and Tris-buffered emulsions, but not in both buffered liposome systems. A mixture of 1 microM lactoferrin and 0.5 microM ferrous ions was a significantly better antioxidant than 1 microM lactoferrin alone in Tris-buffered emulsions and in phosphate-buffered liposomes. Lactoferrin was a prooxidant at 1 microM in phosphate-buffered liposomes and at 15 and 20 microM in Tris-buffered liposomes. Copper was a stronger prooxidant than iron in both buffered emulsions. Lactoferrin decreased the prooxidant effect of iron, but not of copper, in emulsions. The antioxidant or prooxidant activities of lactoferrin depended on the lipid system, buffer, its concentration, the presence of metal ions, and oxidation time.     

Astaxanthin protects against oxidative stress and calcium-induced porcine lens protein degradation

Astaxanthin (ASTX), a carotenoid with potent antioxidant properties, exists naturally in various plants, algae, and seafoods. In this study, we investigated the in vitro ability of ASTX to protect porcine lens crystallins from oxidative damage by iron-mediated hydroxyl radicals or by calcium ion-activated protease (calpain), in addition to the possible underlying biochemical mechanisms. ASTX (1 mM) was capable of protecting lens crystallins from being oxidized, as measured by changes in tryptophan fluorescence, in the presence of a Fenton reaction solution containing 0.2 mM Fe2+ and 2 mM H2O2. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis demonstrated that beta(high)-crystallin was the most vulnerable protein under these conditions of free radical exposure. The proteolysis of lens crystallins induced by calcium ion-activated calpain was also inhibited by ASTX (0.03-1 mM) as determined by daily measurement of the light-scattering intensity at 405 nm for five consecutive days. ASTX at 1 mM was as potent as a concentration of 0.1 mM calpain inhibitor E64 in protecting the oxidative damage/hydrolysis of porcine crystallins. At a concentration of 1 mM, ASTX provided better protection than the endogenous antioxidant glutathione in terms of suppressing calcium-induced turbidity of lens proteins. Thin-layer chromatography analysis indicated that ASTX interacted with calcium ions to form complexes, which we believe interfere with the hydrolysis of lens crystallins by calcium-activated calpain. This in vitro study shows that ASTX is capable of protecting porcine lens proteins from oxidative insults and degradation by calcium-induced calpain.     

Effect of the citrus flavanone naringenin on oxidative stress in rats

Flavonoids are non-nutrient plant phenolic compounds proposed to provide health benefits in humans. The antioxidant and prooxidant effects of the citrus flavanone naringenin have been tested only in vitro. The dose-response effect of naringenin consumption was tested in weanling rats (n=6-8/group) with a 2x4 factorial design using high or low oxidative stress (Hox or Lox, respectively) diets, created by adequate or deficient amounts of vitamin E and selenium, with three increasing naringenin concentrations (30, 60, or 120 mg/kg of diet). Hox compared to Lox rats exhibited reduced growth and liver hypertrophy, which was not prevented by naringenin consumption. Also, Hox rats exhibited severalfold higher liver NAD(P)H:quinone oxidoreductase-1 activity, which was further elevated in proportion to naringenin intake, but this was not sufficient to protect against oxidative stress indicated by higher liver total aldehydes. In addition, dietary naringenin did not affect antioxidant nutrient status or physiological markers of growth under Lox conditions. Thus, dietary naringenin did not exhibit antioxidant or prooxidant effects in vivo in this rat model.     

Lipid peroxidation by "free" iron ions and myoglobin as affected by dietary antioxidants in simulated gastric fluids

Grilled red turkey muscle (Doner Kabab) is a real "fast food" containing approximately 200 microM hydroperoxides, homogenized in simulated gastric fluid and oxidized more rapidly at pH 3.0 than at pH 5.0, after 180 min, producing 1200 and 600 microM hydroperoxides, respectively. The effects of "free" iron ions and metmyoglobin, two potential catalyzers of lipid peroxidation in muscle foods, were evaluated for linoleic acid peroxidation at pH 3.0 of simulated gastric fluid. The prooxidant effects of free iron ions on linoleic acid peroxidation in simulated gastric fluid was evaluated in the presence of ascorbic acid. At low concentrations of ascorbic acid, the effects were prooxidative, which was reversed at high concentrations. In the presence of metmyoglobin, ascorbic acid with or without free iron enhanced the antioxidative effect. Lipid peroxidation by an iron-ascorbic acid system was inhibited totally by 250-500 microM catechin at pH 3.0. The catechin antioxidant effect was determined also in the iron-ascorbic acid system containing metmyoglobin. In this system, catechin totally inhibited lipid peroxidation at a concentration 20-fold lower than without metmyoglobin. The ability of catechin to inhibit lipid peroxidation was also determined at a low pH with beta-carotene as a sensitive target molecule for oxidation. The results show that a significant protection was achieved only with almost 100-fold higher antioxidant concentration. Polyphenols from different groups were determined for the antioxidant activity at pH 3.0. The results show a high antioxidant activity of polyphenols with orthodihydroxylated groups at the B ring, unsaturation, and the presence of a 4-oxo group in the heterocyclic ring, as demonstrated by quercetin.