松栎柱锈菌春孢子多糖体外抗氧化活性

为了研究松栎柱锈菌（Cronartium orientale S.Kaneko）春孢子多糖的体外抗氧化活性,制备了该菌破壁春孢子精制多糖,并对所得春孢子多糖进行了总抗氧化能力、总还原力以及清除二苯代苦味酰基（DPPH·）、清除羟基自由基（·OH）、清除超氧阴离子（O₂^·）的活性进行了测定。体外抗氧化活性分析结果表明：松栎柱锈菌春孢子多糖具有较强的总抗氧化能力;虽然该菌多糖不具有清除多数氧自由基母体（O₂^·）的作用,也不具有清除人工合成自由基DPPH·的作用,但是其能够有效清除对机体破坏力最强的自由基·OH。这表明本文获得的松栎柱锈菌春孢子多糖具有对自由基选择性清除的能力,对其他自由基研究者具有参考价值,可为该菌的开发利用提供参考,也能够为森林有害生物开发利用提供理论依据。     

蛹虫草无性型菌丝体提取液体外抗氧化活性研究

分别测定了蛹虫草无性型菌丝体不同溶剂提取液在不同抗氧化模型中的抗氧化作用。结果表明,各种提取液对二苯基苦味酰基苯肼自由基(·DPPH)和羟自由基(·OH)均有显著的清除作用:在50mg/mL时,去离子水、70%乙醇和70%丙酮提取物对DPPH自由基的清除率分别为89.2%、83.6%和75.9%;70%乙醇提取物在30mg/mL时对·OH自由基的清除率达到100%;在50mg/mL时70%丙酮和去离子水提取物的·OH自由基清除率分别为95.6%和89.6%。在一定浓度下,各提取液对邻苯三酚自氧化也均有抑制作用;不同溶剂提取物的还原能力强弱为:70%乙醇>去离子水>70%丙酮。各种提取物的抗氧化组分不同,提取物浓度与抗氧化性成正相关。     

比色法测定抗坏血酸体系产生的·OH自由基

本文介绍一种用细胞色素c的氧化来比色测定抗坏血酸体系产生的·OH自由基的方法。实验证明,它不受SOD的抑制,仅受硫脲的抑制。所以,该体系可作为筛选抗·OH自由基的清除剂和有效药物的体系。     

唐古特白刺果实花色苷体外清除自由基的活性

为探讨唐古特白刺(Nitraria tangutorun Bobr.)果实花色苷体外清除自由基活性,研究了其对超氧阴离子(O-2·)体系、羟基自由基(·OH)体系和二苯代苦味酰基自由基(DPPH·)体系的清除效果,并与抗坏血酸(vc)进行了比较.结果表明,该花色苷对O-2·、·OH、DPPH·均具有清除作用,且与浓度呈量效关系.对·OH和DPPH·清除效果相对较好,优于Vc.     

κ-卡拉胶邻苯二甲酰基化衍生物的抗氧化活性研究

对κ-卡拉胶进行酸降解得到三种卡拉胶低聚糖,并进一步与苯二甲酰基合成制得三种分子量分别为1450、2520和3430的κ-卡拉胶邻苯二甲酰衍生物(LA、LB和LC)。对产物进行IR表征并对其取代度(DS)进行测定,并检测了产物对羟基自由基.OH、DPPH自由基和过氧化氢的清除活性以及还原能力。结果表明,上述三种κ-卡拉胶邻苯二甲酰衍生物的抗氧化能力强弱顺序依次为:LC>LA>LB,这可能与衍生物的羟基含量、取代基团的性质以及取代度等因素有关。     

芦丁等天然产物清除活性氧自由基O_(?)~-和·OH的ESR研究

本文用促癌剂PMA(phorbol myristate acetate)刺激人多形核白细胞(PMN)呼吸暴发产生的活性氧自由基,Fenton反应产生的羟自由基·OH,光照核黄素和黄嘌呤/黄嘌呤氧化酶体系中产生的超氧阴离子自由基O(?)为模型,用自旋捕集方法研究天然产物芦丁,槲皮素,异槲皮苷和汉防已甲素对活性氧自由基(?)和·OH的清除作用.除汉防已甲素外,其它药物都能很明显地清除PMN呼吸暴发过程中产生的活性氧自由基.芦丁和异槲皮苷对(?)的清除率分别高达78.1%和79.9%,远远大于维生素E(12.7%)的作用.除汉防已甲素外,其它三种药物对·OH的清除作用也大于维生素E.四种天然产物对O(?)和·OH的清除作用都小于维生素C.     

猪苓菌丝细胞活性氧的产生及其种类

用液体发酵的蜜环菌菌丝、菌丝细胞壁及发酵液作为激发子,分别处理猪苓菌丝,均可诱导猪苓菌丝活性氧的产生。活性氧产生量与激发子浓度具相关性。超氧化物歧化酶（SOD）、过氧化氢酶(CAT)、甘露醇均可在一定程度上抑制活性氧的产生,证明活性氧种类包括过氧化氢（H2O2）、羟基自由基（·OH）和超氧根阴离子(O·－2)。Diphenylene iodonium (DPI)能削弱激发子对活性氧的诱导,表明O·－2来源于NADPH氧化酶。     

黑老虎叶总黄酮提取工艺及其抗氧化活性研究

为探讨超声波辅助提取黑老虎叶总黄酮的最佳提取工艺条件及其抗氧化活性,该文以黑老虎叶为研究对象,采用超声波提取法提取黑老虎叶总黄酮,通过单因素试验研究提取时间、乙醇浓度、提取温度、料液比对黑老虎叶总黄酮提取率的影响,在单因素试验的基础上,采用正交试验优化其提取工艺条件,测试了最优条件下提取的黑老虎叶总黄酮对DPPH自由基、·OH自由基及超氧阴离子的清除能力。结果表明:黑老虎叶总黄酮超声辅助提取最佳提取条件为提取时间 35 min、乙醇浓度80%、提取温度50 ℃、料液比1:20 g·mL^-1,最佳条件下提取率为4.83%。抗氧化活性测试结果显示,黑老虎叶总黄酮表现出较好的清除DPPH自由基、·OH自由基及超氧阴离子能力,其抗氧化能力为清除DPPH自由基能力>清除超氧阴离子能力>清除·OH自由基能力。在浓度为0.8 mg·mL^-1时,黑老虎叶总黄酮清除DPPH自由基、·OH自由基及超氧阴离子的能力相当于同浓度下Vc的97.6%、82.1%、95.5%,黑老虎叶总黄酮是天然抗氧化剂的良好来源。上述结果为黑老虎叶活性成分的提取及开发利用提供了理论基础。     

活性氧对苏云金芽孢杆菌伴孢晶体的损伤作用

用SDSPAGE电泳分析和生物测定方法研究了过氧化氢(H2O2)和羟自由基(·OH)对苏云金芽孢杆菌(Bacillus thuringiensis)伴孢晶体的损伤作用。结果表明,这两种活性氧对伴孢晶体均有一定程度的损伤作用,这种损伤作用与活性氧的浓度成正相关,并且·OH对伴孢晶体的损伤作用明显强于H2O2。     

丝素肽的制备及其抗氧化活性研究

本文以废蚕丝为原料,通过脱胶精炼、碱性蛋白酶Alcalase2.4降解后得到丝素肽溶液,并采用超氧自由基(O-·2)体系、羟基自由基(·OH)体系、二苯代苦味酰基自由基(DPPH·)体系考察了不同水解度丝素肽体外清除自由基的活性。结果表明,丝素蛋白溶液在底物浓度5%,加酶量([E]/[S])2%,反应温度60℃,p H值8.5时,通过控制反应时间可获得水解度为0~23.92%的丝素肽,在此范围内丝素肽清除自由基的能力随水解度的提高而不断增强,在水解度为23.92%时,丝素肽对超氧自由基(O-·2)的清除率达73.69%,对羟基自由基(·OH)的清除率为82.49%,对二苯代苦味酰基(DPPH·)的清除率为70.79%,抗氧化活性最好。     

The non-additive contribution of hydroxyl substituents to Akt kinase–apigenin affinity

The natural product apigenin is a flavonoid derivative substituted by three hydroxyl functional groups at positions 4′, 5 and 7 [OH(4′), OH(5) and OH(7)] of the basic flavonoid skeleton, which has shown strong inhibition on the development, proliferation and invasion of tumour cells by binding specifically to Akt kinase to inactivate the Akt signalling pathway. In this study, a typical non-additivity of the three hydroxyl substituents’ contributions to Akt–apigenin binding affinity is demonstrated by combination of four empirical scoring functions, molecular dynamics simulations, molecular mechanics-Poisson–Boltzmann/surface area (MM/PBSA) analyses and hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. It is found that (i) the empirical scoring functions are incapable of properly reflecting the non-additivity feature, which, however, can be well described by the more rigorous MM/PBSA and QM/MM methods, (ii) the hydroxyl group contributions to ligand binding affinity are deviated significantly from linear additive model due to the strong conjugate effect and σ-effect among them, that is, the co-contribution of the three hydroxyl groups is far less than the sum of their individual contributions and (iii) as might be expected, a strong interactive effect is observed for the two adjacent substituents OH(5) and OH(7) as compared with that of distant OH(5) and OH(4′) as well as OH(7) and OH(4′). In addition, the structural basis, energetic property and molecular mechanism of the non-additivity feature are also explored in detail using the natural population analysis and quantum mechanical calculations.     

Spin-trapping studies of hydroxyl radical production involved in lipid peroxidation.

Evidence presented in this report suggests that the hydroxyl radical (OH.), which is generated from liver microsomes is an initiator of NADPH-dependent lipid peroxidation. The conclusions are based on the following observations: 1) hydroxyl radical production in liver microsomes as measured by esr spin-trapping correlates with the extent of NADPH induced microsomal lipid peroxidation as measured by malondialdehyde formation; 2) peroxidative degradation of arachidonic acid in a model OH · generating system, namely, the Fenton reaction takes place readily and is inhibited by thiourea, a potent OH · scavenger, indicating that the hydroxyl radical is capable of initiating lipid peroxidation; 3) trapping of the hydroxyl radical by the spin trap, 5,5-dimethyl-1-pyrroline-1-oxide prevents lipid peroxidation in liver microsomes during NADPH oxidation, and in the model system in the presence of linolenic acid. The possibility that cytochrome P-450 reductase is involved in NADPH-dependent lipid peroxidation is discussed. The optimal pH for the production of the hydroxyl radical in liver microsomes is 7.2. The generation of the hydroxyl radical is correlated with the amount of microsomal protein, possibly NADPH cytochrome P-450 reductase. A critical concentration of EDTA (5 × 10^?5m) is required for maximal production of the hydroxyl radical in microsomal lipid peroxidation during NADPH oxidation. High concentrations of Fe²⁺-EDTA complex equimolar in iron and chelator do not inhibit the production of the hydroxyl radical. The production of the hydroxyl radical in liver microsomes is also promoted by high salt concentrations. Evidence is also presented that OH radical production in microsomes during induced lipid peroxidation occurs primarily via the classic Fenton reaction.     

Spin trapping evidence for myeloperoxidase-dependent hydroxyl radical formation by human neutrophils and monocytes.

Using the electron spin resonance/spin trapping system, 4-pyridyl 1-oxide N-tert-butylnitrone (4-POBN)/ethanol, hydroxyl radical was detected as the alpha-hydroxyethyl spin trapped adduct of 4-POBN, 4-POBN-CH(CH3)OH, from phorbol 12-myristate 13-acetate-stimulated human neutrophils and monocytes without the addition of supplemental iron. 4-POBN-CH(CH3)OH was stable in the presence of a neutrophil-derived superoxide flux. Hydroxyl radical formation was inhibited by treatment with superoxide dismutase, catalase, and azide. Treatment with a series of transition metal chelators did not appreciably alter 4-POBN-CH(CH3)OH, which suggested that hydroxyl radical generation was mediated by a mechanism independent of the transition metal-catalyzed Haber-Weiss reaction. Kinetic differences between transition metal-dependent and -independent mechanisms of hydroxyl radical generation by stimulated neutrophils were demonstrated by a greater rate of 4-POBN-CH(CH3)-OH accumulation in the presence of supplemental iron. Detection of hydroxyl radical from stimulated monocyte-derived macrophages, which lack myeloperoxidase, required the addition of supplemental iron. The addition of purified myeloperoxidase to an enzymatic superoxide generating system resulted in the detection of hydroxyl radical that was dependent upon the presence of chloride and was inhibited by superoxide dismutase, catalase, and azide. These findings implicated the reaction of hypochlorous acid and superoxide to produce hydroxyl radical. 4-POBN-CH(CH3)OH was not observed upon stimulation of myeloperoxidase-deficient neutrophils, whereas addition of myeloperoxidase to the reaction mixture resulted in the detection of hydroxyl radical. These results support the ability of human neutrophils and monocytes to generate hydroxyl radical through a myeloperoxidase-dependent mechanism.     

ADP-iron as a Fenton reactant: radical reactions detected by spin trapping, hydrogen abstraction, and aromatic hydroxylation

A mixture of ADP, ferrous ions, and hydrogen peroxide (H2O2) generates hydroxyl radicals (OH) that attack the spin trap DMPO (5,5-dimethyl-pyrollidine-N-oxide) to yield the hydroxyl free radical spin-adduct, degrade deoxyribose and benzoate with the release of thiobarbituric acid-reactive material, and hydroxylate benzoate to give fluorescent products. Inhibition studies, with scavengers of the OH radical, suggest that the behavior of iron-ADP in the reaction is complicated by the formation of ternary complexes with certain scavengers and detector molecules. In addition, iron-ADP reacting with H2O2 appears to release a substantial number of OH radicals free into solution. During the generation of OH radicals the ADP molecule was, as expected, damaged by the iron bound to it. Damage to the iron ligand in this way is not normally monitored in reaction systems that use specific detector molecules for OH radical damage. Under certain reaction conditions the ligand may be the major recipient of OH radical damage thereby leading to the incorrect assumption that the iron ligand is a poor Fenton reactant.     

Sialic acid is an essential moiety of mucin as a hydroxyl radical scavenger

In this work, we examined the antioxidant role of mucin, a typical sialic acid containing high-molecular weight glycoprotein. The function of mucin as a hydroxyl radical (.OH) scavenger was characterized using bovine submaxillary gland mucin (BSM). Non-treated BSM effectively protected DNA from the attack of .OH; however, desialylated BSM lost this potential. Moreover, we estimated the scavenging effects of BSM against .OH generated by UV irradiation of hydrogen peroxide using ESR analysis. Our results indicate that BSM has .OH scavenging ability the and sialic acid in mucin is an essential moiety to scavenge .OH.     

Intramolecular hydrogen bonds and conformation of the lincomycin molecule in organic solvents

IR spectra (1600-1800 and 3000-3650 cm-1) of lincomycin base solutions in inert (CCl4 and C2Cl4), proton acceptor (dioxane, dimethylsulfoxide and triethyl amine) and proton donor (CHCl3, CD3OD and D2O) solvents were studied. Analysis of the concentration and temperature changes in the spectra revealed that association in lincomycin in the inert solvents was due to intramolecular hydrogen linkage involving amide and hydroxyl groups. Disintegration of the associates after the solution dilution and temperature rise was accompanied by formation of intramolecular bonds stabilizing the stable conformation structure of the lincomycin molecule. The following hydrogen linkage in the conformation was realized: NH...N (band v NH...N at 3340 cm-1), OH...O involving the hydroxyl at C-7 and O atoms in the D-galactose ring (band v OH...O at 3548 cm-1), a chain of the hydrogen bonds OH...OH...OH in the lincomycin carbohydrate moiety (band v OH...O at 3593 cm-1 and v OH of the end hydroxyl group at 3625 cm-1). Bonds NH and C-O of the amide group were located in transconformation. Group C-O did not participate in the intramolecular hydrogen linkage.     

Relative importance of cellular uptake and reactive oxygen species for the toxicity of alloxan and dialuric acid to insulin-producing cells

The diabetogenic agent alloxan is selectively accumulated in insulin-producing cells through uptake via the GLUT2 glucose transporter in the plasma membrane. In the presence of intracellular thiols, especially glutathione, alloxan generates "reactive oxygen species" (ROS) in a cyclic reaction between this substance and its reduction product, dialuric acid. The cytotoxic action of alloxan is initiated by free radicals formed in this redox reaction. Autoxidation of dialuric acid generates superoxide radicals (O(2)(*-)) and hydrogen peroxide (H(2)O(2)), and finally hydroxyl radicals ((*)OH). Thus, while superoxide dismutase (SOD) only reduced the toxicity, catalase, in particular in the presence of SOD, provided complete protection of insulin-producing cells against the cytotoxic action of alloxan and dialuric acid due to H(2)O(2) destruction and the prevention of hydroxyl radical ((*)OH) formation, indicating that it is the hydroxyl radical ((*)OH) which is the ROS ultimately responsible for cell death. After selective accumulation in pancreatic beta cells, which are weakly protected against oxidative stress, the cytotoxic glucose analogue alloxan destroys these insulin-producing cells and causes a state of insulin-dependent diabetes mellitus through ROS-mediated toxicity in rodents and in other animal species, which express this glucose transporter isoform in their beta cells.     

On the impossibility of OH radical formation from the hydroxyl ion in water

The enthalpy and free energy of the formation of OH radical from the hydroxyl ion in water have been calculated. It is shown that this reaction cannot take place.     

Cobalt(II) ion as a promoter of hydroxyl radical and possible 'crypto-hydroxyl' radical formation under physiological conditions. Differential effects of hydroxyl radical scavengers

Co(II) ions react with hydrogen peroxide under physiological conditions to form a 'reactive species' that can hydroxylate aromatic compounds (phenol and salicylate) and degrade deoxyribose to thiobarbituric-acid-reactive material. Catalase decreases the formation of this species but superoxide dismutase or low concentrations of ascorbic acid have little effect. EDTA, present in excess over the Co(II), can accelerate deoxyribose degradation and aromatic hydroxylation. In the presence of EDTA, deoxyribose degradation by the reactive species is inhibited competitively by scavengers of the hydroxyl radical (.OH), their effectiveness being related to their second-order rate constants for reaction with .OH. In the absence of EDTA the scavengers inhibit only at much higher concentrations and their order of effectiveness is changed. It is suggested that, in the presence of EDTA, hydroxyl radical is formed 'in free solution' and attacks deoxyribose or an aromatic molecule. In the absence of EDTA, .OH radical is formed in a 'site-specific' manner and is difficult to intercept by .OH scavengers. The relationship of these results to the proposed 'crypto .OH' radical is discussed.     

Interactions of Hydroxyl Radicals with Tris (Hydroxymethyl) Aminomethane and Good's Buffers Containing Hydroxymethyl or Hydroxyethyl Residues Produce Formaldehyde

The production of formaldehyde from tris(hydroxymethyl) aminomethane(Tris) by interaction with hydroxyl radicals(.OH) was studied, since the reaction mixture from the Fenton reaction performed in Tris/HCl buffer was found to be color-developed by colorimetric determination of formaldehyde. The absorption spectrum of chromogens was identical to that of authentic formaldehyde. Color development, which required the presence of Tris, hydrogen peroxide and cupric ions in the Fenton reaction mixture, was inhibited by the addition of hydroxyl radical scavengers such as glucose or hyaluronic acid. These results indicated that formaldehyde was produced when Tris interacted with ·OH. With structures similar to Tris, Good's buffers were also found to produce formaldehyde by interaction with ·OH. Analysis of formaldehyde derived from these buffers may provide a simple and convenient assay for detecting ·OH generation. In evaluating effects of ·OH on the biological system in Tris/HCl buffer or certain Good's Buffers, ·OH loss may be due to interactions of ·OH with these buffers. The formaldehyde produced as a result of such interactions may affect biological systems.