不同提取方法对豫产白花蛇舌草多糖及抗氧化活性的影响

为评价不同提取方法对河南地区白花蛇舌草粗多糖的得率、总碳水化合物含量、杂质含量和抗氧化活性的影响,本实验分别以传统热水提取法、超声波辅助提取法、纤维素酶提取法从该药材中提取多糖。同时测定了粗多糖中总碳水化合物含量、杂质含量(总蛋白和总酚含量)以及羟基自由基清除能力、DPPH自由基清能力、还原力等抗氧化能力。实验结果显示三种提取法所得粗多糖相对于原药材的得率分别为6.77±0.09%、6.37±0.01%和4.81±0.90%;粗多糖中的总碳水化合物含量分别为289.14±1.33、246.83±2.33和278.92±2.92mg/g。粗多糖中总蛋白含量分别为28.59±2.21、23.26±2.43和4.96±0.18mg/g,总酚类成分含量分别为5.9±0.08、5.31±0.40和2.82±0.07mg/g。抗氧化活性实验结果表明,三种粗多糖的羟基自由基清除能力呈现出浓度依赖性,且均在3.0mg/mL时达到最大,对应最大清除率分别为89.13%、85.87%和74.71%。另外,三种粗多糖的DPPH自由基清除率分别在0.75~3.0mg/mL的浓度区间内达到最大并保持稳定,对应清除率的稳定区间分别为77.02%~77.90%、77.06%~77.96%和83.16%~85.73%。综上所述,以上三种不同提取方法对白花蛇舌草多糖的品质影响较大,可以为豫产白花蛇舌草的品质鉴别、多糖质量控制和开发利用提供依据。     

印度块菌提取物抗氧化活性的研究

对印度块菌Tuber indicum子实体的提取物,包括55%乙醇粗提物(ECE)、石油醚提取物(PEF)和乙酸乙酯提取物(EAF)的清除DPPH自由基和羟基自由基能力、铁离子鳌合能力以及各提取物的总酚含量等进行了研究和测定,结果显示,3种提取物的清除自由基能力和铁离子鳌合能力具有显著差异(P0.05);ECE对DPPH自由基的清除活性最高,其EC50值为1.61g/L;EAF对羟基自由基及铁离子表现出较强的清除或螯合的能力,其EC50值分别为3.31g/L和0.70g/L;EAF的总酚含量(2.964mg GAE/g提取物)最高,其次是ECE,总酚含量为(2.618mg GAE/g提取物);PEF的清除自由基和铁离子鳌合能力较差,其总酚含量也最低(1.124mg GAE/g提取物);总酚含量与印度块菌提取物清除自由基以及鳌合铁离子的能力密切相关。     

几种药用植物总多酚含量及其抗氧化活性比较

测定了5种药用植物的总多酚含量,对提取液的抗氧化活性和自由基清除能力进行了比较。结果表明:金银花的总多酚含量最高为0.085 8 mg/mL,5种药用植物多酚化合物浓度顺序为:金银花连翘茵陈海金沙过路黄。海金沙多酚提取液的抗氧化活性最强,60℃反应7h,在芝麻油和膏霜化妆品中添加海金沙总多酚提取液,其过氧化值比添加BHT分别低18.27%和15.89%,比不加抗氧剂分别低20.85%和17.97%。连翘的羟基自由基清除率最大,是BHT的2.99倍。     

新疆准噶尔柳花化学成分分析及抗氧化活性的研究

目的：探索准噶尔柳花化学成分的组成及其花茶适宜的日常饮用方式，揭示准噶尔柳花的抗氧化能力。方法：以6月、7月、8月采摘的准噶尔柳花为研究材料，首先对准噶尔柳花的水分、可溶性总糖、总酚、总黄酮、维生素C、可溶性蛋白含量进行测定，再通过单因素实验确定不同冲泡因素对柳花溶出物的影响，最后以1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除率和总抗氧化能力评价柳花提取物体外抗氧化活性。结果：准噶尔柳花的主要化学成分由高到低依次为可溶性蛋白100.03 mg/g、可溶性总糖87.20 mg/g、总黄酮52.61 mg/g、总酚26.57 mg/g、维生素C 6.42 mg/g；准噶尔柳花茶日常适宜的冲泡温度为90℃、时间10 min、料液比150 mL/g、冲泡次数2次；DPPH自由基清除率最高为6月柳花91.01%，总抗氧化能力最高为6月柳花27.63 U/mL。结论：准噶尔柳花具有丰富的营养成分和较强的抗氧化能力，并且6月采摘的柳花抗氧化能力更强，因此新疆准噶尔柳花是一种具有较强开发利用价值的新疆野生植物资源。     

小檗果实中酚类化合物含量测定及其抗氧化活性研究

本试验采用UV法和HPLC法测定小檗果实中酚类化合物的含量,通过DPPH自由基清除率对酚类化合物进行抗氧化活性研究。UV法测得小檗果实总酚含量为3.858 mg/g,HPLC法测得小檗果实中绿原酸、芦丁、槲皮素、山奈酚平均含量分别为0.980 22%、0.006 23%、0.000 44%、0.001 51%;测得DPPH自由基清除率K为85.59%。HPLC法分析速度快,重复性好,能较准确测定小檗果实中总酚含量,且其中酚类化合物具有较强的抗氧化活性,小檗果实可作为天然抗氧化原料开发应用研究。     

李氏块菌提取物抗氧化活性的研究

对李氏块菌Tuber liyuanum子实体的不同提取物,包括甲醇提取物(ME)、乙醇提取物(EE)、丙酮提取物(AE)、正丁醇提取物(BAE)和乙酸乙酯提取物(EAE),进行清除DPPH自由基、羟基自由基、超氧阴离子自由基、铁离子螯合能力、还原力以及总酚含量的测定和研究,发现ME对DPPH自由基、超氧阴离子自由基清除及还原力活性最高,EC50值分别为23.37mg/m L、11.65mg/m L和24.47mg/m L;EE对羟基自由基清除和铁离子螯合能力活性最高,EC50值分别为7.24mg/m L和小于0.5mg/m L;ME的总酚含量最高(3.08mg GAE/g提取物),其次是EE(1.34mg GAE/g提取物),提取物总酚含量与抗氧化活性呈现出一定的正相关性。     

新疆不同红枣品种的总酚含量和抗氧化能力比较

为了研究新疆红枣的遗传多样性,寻找抗氧化能力较强的功能性红枣品种,为保护利用红枣资源提供基础,用Folin-Ciocalteau法测定了7个品种红枣甲醇提取液中的总酚含量,采用体外法评价了红枣甲醇提取物的总抗氧化能力及对DPPH·和羟基自由基(·OH)的清除能力。结果表明:7个参试红枣品种都具有强抗氧化性,并且大多数品种间差异显著,其中哈密大枣、骏枣、灰枣的抗氧化能力较强;不同品种间的总酚含量差异较大,变化值为751.25 mg/100g,其中哈密大枣中含量最高是1 837.48 mg/100g,新疆小圆枣的总酚含量最低为1 086.23 mg/100g;相同品种中果肉和果核间总酚含量差异较大,果肉总酚含量约为果核的6倍。各种抗氧化指标与总酚含量呈显著正相关,酚类物质是红枣抗氧化的重要物质基础之一。     

益智的抗氧化作用

本文探讨了益智(Alpinia oxyphylla Miquel)超临界CO2提取物及其渣的水提物、正丙醇提取物和乙酸乙酯提取物的抗氧化作用,测定了总酚含量、黄酮含量、抗氧化力、还原能力、DPPH清除率.结果表明,益智超临界CO2提取物和正丙醇提取物的总酚含量最高,均为5.53%,乙酸乙酯提取物的总酚含量为4.04%,水提物总酚含量最低,为O.89%.抗氧化力与酚含量相关(R2=0.703).四种提取物中黄酮含量顺序为:乙酸乙酯提取物(6.29%)＞丙醇提取物(5.81%)＞水提取物(4.85%)＞超临界CO2提取物(4.70%).在还原能力、清除DPPH自由基和羟自由基方面,乙酸乙酯提取物表现出了很强的抗氧化能力,呈现剂量依赖关系.     

硅对镉胁迫下红树白骨壤酚类代谢的影响

采用水培方式研究硅对重金属镉胁迫下白骨壤幼苗酚类代谢影响,探讨硅对白骨壤耐受重金属胁迫的影响,结果表明,低浓度镉胁迫(Cd 0.5 mg·L-1)下,随着外源硅的增加,叶片和根系丙二醛含量降低,但Si 50 mg·L-1处理中各器官总酚含量最高,硅对各器官单宁含量影响不显著;高浓度镉(Cd 5 mg·L-1)导致白骨壤叶片和根系膜脂过氧化,使得丙二醛含量显著高于对照,外源硅的添加显著降低了白骨壤叶片和根系丙二醛积累,Si 100mg·L-1显著增加了各器官中总酚含量和单宁含量,根系丙二醛含量与根系总酚含量呈显著负相关,叶片丙二醛含量与叶片单宁含量呈显著负相关,表明硅显著刺激高镉胁迫下白骨壤植物组织中酚类物质代谢,增加根系和叶片的抗氧化性,进而增加白骨壤对镉的耐受性。     

香露兜叶提取物抗氧化与辐射效应影响

为研究辐射效应对香露兜叶提取物的影响,本研究中利用超声波萃取技术进行香露兜叶的萃取,再以总多酚、总类黄酮、DPPH自由基清除率、亚铁离子螯合等非酵素系统进行香露兜叶提取物抗氧化能力的评估。以过氧化氢诱发氧化压力的毒性效应测试香露兜叶提取物细胞内抗氧化的能力,再以5.5戈雷(Gy)的辐射剂量照射,用MTT分析方法测定细胞的存活率,评估香露兜叶提取物对辐射效应的影响。研究结果显示香露兜叶总多酚含量为(139.89±0.43) GAE mg/g,总类黄酮含量为(44.24±0.61) RE mg/g,DPPH与亚铁离子螯合的IC50分别为(5.87±0.02) mg/mL和(15.3±0.07) mg/mL。在H2O2对细胞伤害的实验中,香露兜叶提取物对于肝癌细胞(HepG2)有显著(p0.05)的毒杀作用,且细胞存活率会随着提取物浓度增加而增高杀死细胞的比率。萃取液浓度分别为25 g/mL、250 g/mL、1 000 g/mL和1 500 g/mL时,经过72 h培养后,细胞存活率分别为64.7%、50.4%、37.5%和30.6%。低浓度香露兜提取物对于HepG2细胞有辐射增敏作用,以浓度为25 g/mL的香露兜提取物合并5.5 Gy的辐射剂量照射,发现其存活率由单独以5.5 Gy辐射剂量照射的54.58%降至12.11%。综合以上的结果,香露兜叶提取物具有不错的抗氧化能力,并且有不错的辐射效应影响能力。     

Scavenging of reactive oxygen species by the plant phenols genistein and oleuropein.

The plant-derived phenolic compounds genistein and oleuropein are known to exhibit several biological properties, many of which may result from their antioxidant and free radical scavenger activity. In this paper we report the results of a complex study of antioxidant activity of genistein and oleuropein, using electron spin resonance (ESR), chemiluminescence, fluorescence and spectrophotometric techniques. Different reaction systems were applied to study the inhibitory effect of the phenolic compounds studied: (a) the potassium superoxide/18-crown-6 dissolved in DMSO system, which generates superoxide radical (O(2).(-)) and hydrogen peroxide (H(2)O(2)); (b) the Co(II)-EDTA-H(2)O(2) system (the Fenton-like reaction), which generates hydroxyl radical (HO.); (c) 2,2'-azobis(2-amidino-propane)dichloride (AAPH) as the peroxyl radical (ROO.) generator, and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical test. Results showed that genistein and oleuropein decreased the chemiluminescence sum from the O(2).(-) generating system, an inhibitory effect that was dependent on their concentration. These compounds also reacted with ROO radicals and they showed activity about two-fold greater than the standard Trolox. The antioxidant effects were studied at different concentrations and reflected in protection against the fluorescence decay of beta-phycoerythrin (beta-PE), due to ROO. attack on this protein. Using the Fenton-like reaction and the spin trap agent 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), the phenolic compounds examined were found to inhibit DMPO-.OH radical formation in the range 10-90% at concentrations of 0.1 mmol/L to 2 mmol/L. Furthermore, these compounds also inhibited HO.-dependent deoxyribose degradation; about 20% and 60% inhibitions were observed in the presence of 0.5 mmol/L genistein and oleuropein, respectively. It was also demonstrated that genistein had a weaker DPPH radical scavenging activity than oleuropein. Our results confirm good scavenging activity towards O(2).(-), HO. and ROO. and the antioxidant effect of genistein and oleuropein.     

氧化胁迫对Alkalibacterium sp. F26防御酶和辅因子的影响

将一株能够高产过氧化氢酶的低度嗜盐嗜碱茵Alkalibacterium sp.F26作为模式微生物,采用高效液相色谱技术测定胞内代谢物浓度,研究氧化胁迫对其防御酶活性和辅因子的影响.研究结果表明:相比低浓度H2O2(＜1 mmol/L)胁迫,此菌株在高浓度H2O2(＞1 mmol/L)胁迫下的应答表现曼为明显:经3 mmol/L H2O2胁迫后胞内CAT酶活为106.54 U/mg protein,是对照产量的1.76倍;ATP浓度则从对照浓度20.55 μmol/L下降到17.80 μmol/L;NAD 浓度自对照样品的69.89 μmol/L减少至31.77 μmol/L.由于ATP和NAD 浓度的减少,相比未经过H2O2胁迫菌体.细胞能荷值EC从0.77降低至0.68,NADH/NAD 则从0.08增加至0.41.然而,这种应答机制在细胞受到低浓度H2O2的胁迫后并不明显:除发现100 μmol/L H2O2能够导致细胞防御机制的激活而使胞内ATP浓度相比对照有所增加的情况外,经50 μmol/L和500 μmol/L H2O2胁迫后胞内ATP水平从对照的22.69 μmol/L只下降到22.38 μmol/L和13.70 μmol/L;并且此种胁迫条件下NADH浓度变化也不显著.     

棘孢木霉菌对钠胁迫的生理响应机制

【背景】棘孢木霉菌制剂被广泛应用于生物防治和次生盐渍化土壤的微生物修复,但是关于棘孢木霉在盐渍化胁迫条件下生长的耐盐机理及其富集盐离子的能力尚缺乏深入研究。【目的】揭示一株耐盐棘孢木霉菌(Trichodermaasperellum)CTCCSJ-W-SBW10264(T264)对钠胁迫的生理响应机制及其对钠离子的吸附和累积特性。【方法】设计梯度浓度钠胁迫培养实验,采集不同培养时期的菌丝样本,测定细胞氧化损伤相关指标H_2O_2和丙二醛(Malondialdehyde,MDA)的含量及细胞抗氧化相关酶的活性变化。【结果】钠胁迫实验表明,棘孢木霉T264能够在1.22mol/L的钠胁迫环境中生存,在低于0.25 mol/L的钠胁迫下其生长不会被明显抑制。细胞氧化损伤及氧化损伤响应相关指标的研究结果表明,培养液中钠离子浓度越高,棘孢木霉的膜系统氧化水平(MDA含量)越高,而且随着细胞中MDA和H_2O_2的累积,细胞抵御氧化损伤相关酶的活性也有明显提高,在钠盐处理24 h后,0.5、1.0和1.22 mol/L的钠离子胁迫分别使过氧化物酶(Peroxidase,POD)、超氧化物歧化酶(SuperoxideDismutase,SOD)和过氧化氢酶(Catalase,CAT)活性达到峰值,依次为36.66、3.34和233.3 U/mg。钠离子吸附和累积特性实验结果表明,棘孢木霉T264的菌丝对钠离子有强的吸附能力。在0.05 mol/L的钠离子环境中培养72 h后,菌丝表面钠离子吸附量为1 347.6 mg/g,菌丝内部钠离子累积量为218.6 mg/g,木霉菌菌丝通过菌丝表面吸附和菌丝内部累积对培养液中钠离子的去除率达到32%。【结论】T264的抗氧化损伤相关酶在其耐受钠离子胁迫过程中发挥重要作用,菌株T264对高浓度钠离子有强适应性,而且对环境中钠离子有高效的吸附和累积作用。     

A new strategy to enhance glutathione production by multiple H2O2 induced oxidative stresses in Candida utilis

Effect of H(2)O(2)-induced oxidative stress on glutathione (GSH) production in Candida utilis was investigated. Based on the results that H(2)O(2) can effectively stimulate GSH accumulation but inhibit cell growth simultaneously, a novel strategy of multiple H(2)O(2) stresses with different concentrations (1 mmol/L at 4h, 2 mmol/L at 8h, and 4 mmol/L at 12h) were developed to maximize GSH production. As a result, a maximal GSH yield of 218 mg/L was achieved and a corresponding intracellular GSH content was 2.15%, which were 54.6% and 58.1% higher than the control. By further applying this strategy to 7 L fermentor, GSH yield and intracellular GSH content were 328 mg/L and 2.30%. Moreover, increased activities of catalase (CAT) and GSH reductase (GR) indicated that GSH and CAT were directly involved in protecting cell against oxidative stress by H(2)O(2).     

Accumulation of antioxidant phenolic constituents in submerged cultures of Inonotus obliquus

Phenolic compounds produced by sclerotia of Inonotus obliquus are the active constituents responsible for antioxidant activities. In this study, I. obliquus was grown in a continuously stirred tank reactor (CSTR) to explore how it accumulates phenolic compounds in different culture media and whether these compounds possess antioxidant activities. Phenolic compounds produced by I. obliquus in the control medium consisted of melanins, flavonoids, polyphenols and small phenolics. Their accumulation was affected by adding H(2)O(2) to the medium, where increased levels of total intracellular phenols (TIP) and melanins, but less total extracellular phenol (TEP) occurred. Simultaneous exposure to H(2)O(2) and arbutin resulted in a further increase in TIP production and reduced accumulation of TEP. Both TIP and TEP obtained at different culture ages and media were active in scavenging superoxide anion and DPPH radicals. Therefore, production of phenolic compounds by I. obliquus is enhanced by imposing oxidative stress, which might allow it to be exploited as a reliable source of pharmaceutically important phenolic compounds.     

一氧化氮对桦褐孔菌抗氧化酚类化合物积累的影响

本研究探讨了NO对深层培养的桦褐孔菌积累抗氧化酚类化合物的影响。在桦褐孔菌的培养基中分别加入0.01mmol/L、0.1mmol/L以及1mmol/L的NO供体硝普钠,并测定总酚的含量及其抗氧化活性。发酵产物的抗氧化活性以清除DPPH和羟自由基活力表示。加入0.1mmol/L的硝普钠可使桦褐孔菌胞内外酚类化合物分别达到最高水平67mg/g和677mg/L。不同培养时期产生的胞内外酚类化合物都具有抗氧化活性,尤其是添加0.1mmol/L硝普钠的菌丝体。因此NO可用于上调深层发酵培养的桦褐孔菌酚类化合物的积累。     

茉莉酸和双氧水对斜生褐孔菌一株黑色素缺失突变体产生酚类化合物的影响

黑色素合成是桦褐孔菌Phaeoporus obliquus次生代谢过程中的一个重要事件。黑色素的过多合成势必影响其他酚类的积累。本文以茉莉酸和双氧水为调控因子,开展了该突变体深层发酵过程酚类化合物积累研究。正常培养条件下,每升发酵液积累酚类化合物最高可达317.45mg/L,其中包括苯甲酸衍生物(45.3mg/L),黄酮苷元(74.4mg/L),黄酮苷(160mg/L)以及硬毛素类化合物(18.5mg/L)。加入茉莉酸后,总酚最大值增加到620.45mg/L,其中苯甲酸衍生物增加到230.26mg/L,黄酮苷元增加到166.86mg/L,而黄酮苷和硬毛素类化合物却分别降低到98.5mg/L和7.1mg,双氧水的加入没有提高总酚的最大积累(315.69mg/L)。因此,与野生型相比,黑色素缺失使总酚的积累有所增加,随茉莉L酸的加入总酚积累最大值增加近一倍。虽然双氧水的加入提高了苯丙氨酸裂解酶活性,但总酚的积累却没有增加,这可能是由于一部分酚类化合物在抗双氧水氧化胁迫过程中被消耗。     

拟细羽束梗孢发酵菌丝体中自由基清除活性成分分析

用二苯基苦基苯肼(DPPH)自由基法,首次对拟细羽束梗孢(Isaria gracilioides RCEF3 279)菌丝体的不同溶剂提取物进行了自由基清除活性的定性定量分析,发现其甲醇提取物具有较强的清除自由基活性,当菌丝浓度为10 g/L时,其甲醇提取物的自由基清除率达到了92.4％±0.3％.DPPH自显影-薄层...     

Anti-oxidant and pro-oxidant behaviour of bucillamine.

Bucillamine (BUC) is used clinically for the treatment of rheumatoid arthritis. Some of the pharmacological action of BUC has been reported as being dependent on the production of reactive oxygen species (ROS). In this paper the reactivity of BUC with superoxide anion radical (O(2) (*-)) generated from potassium superoxide/18-crown-6 ether dissolved in DMSO, hydroxyl radical (HO(*)) produced in the Cu(2+)-H(2)O(2) reaction, peroxyl radical (ROO(*)) from 2,2'-azobis (2-amidino-propane) dichloride decomposition, and singlet oxygen ((1)O(2)) from a mixture of alkaline aqueous H(2)O(2) and acetonitrile, have been investigated. Chemiluminescence, fluorescence, electron paramagnetic resonance (EPR) spin-trapping techniques and the deoxyribose and oxygen radical absorbance capacity towards ROO(*) (ORAC(ROO)) assays were used to elucidate the anti- and pro-oxidative behaviours of BUC towards ROS. The results indicated that BUC efficiently inhibited chemiluminescence from the O(2) (*-)-generating system at relatively high concentrations (0.5-2 mmol/L); however, at lower concentrations (<0.5 mmol/L) the drug enhanced light emission. The behaviour of BUC was correlated with a capacity to decrease the chemiluminescence signal from the Cu(2+)-H(2)O(2) system; scavenging HO(*) was effective only at high concentrations (1-2 mmol/L) of the drug. Bucillamine also prevented deoxyribose degradation induced by HO(*) in a dose-dependent manner, reaching maximal inhibition (24.5%) at a relative high concentration (1.54 mmol/L). Moreover, BUC reacts with ROO(*); the relative ORAC(ROO) was found to be 0.34 micromol/L Trolox equivalents/micromol sample. The drug showed quenching of (1)O(2)-dependent 2,2,6,6-tetramethylpiperidine-N-oxide radical formation from 2,2,6,6-tetramethyl-piperidine (e.g. 90% inhibition was found at 1 mmol/L concentration). The results showed that BUC may directly scavenge ROS or inhibit reactions generating them. However, the drug may have pro-oxidant activity under some reaction conditions.     

The antioxidant activity of glucosamine hydrochloride in vitro

The antioxidant potency of chitin derivative-glucosamine hydrochloride was investigated employing various established in vitro systems, such as superoxide (O2*-)/hydroxyl (*OH)-radical scavenging, reducing power, and ferrous ion chelating potency. As expected, we obtained several satisfying results, as follows: first, glucosamine hydrochloride had pronounced scavenging effect on superoxide radical. For example, the O2*- scavenging activity of glucosamine hydrochloride was 83.74% at 0.8 mg/mL. Second, the *OH scavenging activity of glucosamine hydrochloride was also strong and was about 54.89% at 3.2 mg/mL. Third, the reducing power of glucosamine hydrochloride was more pronounced. The reducing power of glucosamine hydrochloride was 0.632 at 0.75 mg/mL. However, ferrous ion-chelating potency was soft. Furthermore, ferrous ion-chelating potency, the scavenging rate of radical, and the reducing power of glucosamine hydrochloride increased with their increasing concentration, and they were concentration dependent. The multiple antioxidant activity of glucosamine hydrochloride was evident as it showed considerable reducing power, superoxide/hydroxyl-radical scavenging ability. These in vitro results suggest the possibility that glucosamine hydrochloride could be effectively employed as an ingredient in health or functional food, to alleviate oxidative stress.