硒多糖的抗氧化活性及与过氧化氢酶协同作用的研究

对自行培养的富硒蛹虫草中提取的有机硒多糖进行抗氧化活性研究.采用循环伏安法(CV),在铂电极表面构筑支撑磷脂双层膜 (Supported bilayer lipid membrane, s-BLM),以0.1 mol/L KCl-1.0 mmol/L K3Fe(CN)6-1.0 mmol/L K4Fe(CN)6为探针溶液,利用Fenton体系中Fe2+和H2O2反应产生的羟自由基对模拟生物膜的损伤程度考察有机硒多糖的生物活性.结果表明:羟自由基使s-BLM的通透性增强,造成膜体损伤;而在硒多糖存在下,可使羟自由基对膜的这种损伤受到抑制,硒多糖能够清除自由基保护生物膜;采用紫外-可见(UV-Vis)吸收光谱法比较研究不同浓度的过氧化氢酶与硒多糖对羟自由基的清除率.结果表明:硒多糖对羟自由基的清除效果好于过氧化氢酶,且当体系中硒多糖和过氧化氢酶共存时,二者对羟自由基的清除效果具有协同作用.进一步讨论了硒多糖抗氧化活性的可能机理,为有机硒多糖在保护生物膜、抗氧化活性方面的开发与利用提供科学的理论依据.     

Enterobacter Cloacae Z0206细菌胞外富硒多糖的抗氧化活性

利用产多糖菌Enterobacter cloacae Z0206(E.cloacaeZ0206)的深层发酵法制备了E.cloacae Z0206细菌富硒多糖;测定了其还原能力和清除1,1-diphenyl-2-picrylhydrazyl(DPPH)自由基、超氧阴离子及羟自由基的能力.结果表明,通过深层富硒发酵、醇沉离心等制备富硒多糖SEPS的产量为9.28g/L,富硒量为2.314mg/g;E.cloacae Z0206富硒多糖对DPPH自由基和羟自由基具有较好的清除作用,在浓度为5g/L时对DPPH自由基和羟自由基的清除率分别为80.35%和84.26%,并具有较强的还原能力,但其对超氧阴离子自由基的清除能力较差.     

石墨烯纳米载药体系的制备及对人口腔鳞癌细胞杀伤效果

利用化学氧化还原法制备了氧化石墨烯,进一步超声破碎剥离,得到纳米氧化石墨烯,并对其进行聚乙二醇(PEG)的功能化修饰后载药顺铂。 采用扫描电子显微镜(SEM)、紫外-可见吸收光谱(UV-Vis)、傅立叶变换红外光谱(FTIR)对石墨烯纳米载药体系进行表征,细胞存活率实验(MTT)法检验石墨烯纳米载药体系对人口腔鳞癌(KB)细胞的杀伤作用。 结果表明,石墨烯纳米载药体系对顺铂的负载率为42.4%,聚乙二醇修饰后可以降低纳米氧化石墨烯的细胞毒性并提高生物相容性,对KB细胞具有双重的杀伤作用,为纳米氧化石墨烯在肿瘤治疗的临床应用提供了理论依据。     

富硒蛹虫草的抗氧化活性研究

采用现代分离技术从富硒蛹虫草中提取硒多糖,然后以苯酚-硫酸法、电感耦合等离子体质谱（ICP—MS）法对硒多糖中多糖含量和硒含量进行测定;在Fenton体系的最佳实验条件下,研究了硒多糖对羟自由基的清除作用及硒多糖对支撑磷脂双层膜（Supported Bilayer Lipid Membranes,s—BLM）的保护作用。结果表明：硒多糖清除羟自由基的效果比与之相同浓度的无机硒、多糖都要明显。其中硒多糖对羟自由基的最高清除率可达38．02％,硒多糖具有很强的抗脂质过氧化、保护支撑磷脂双层膜的作用。     

具有谷胱甘肽过氧化物酶(GPx)活力的含硒四肽(SecRGD)

以精氨酸-甘氨酸-天冬氨酸(RGD)序列为基础, 在N-端引入硒代半胱氨酸(Sec)设计了SecRGD序列模拟谷胱甘肽过氧化物酶(GPx), 利用Fmoc固相合成法合成了SecRGD. 采用ESI-MS质谱和氢化物原子荧光光谱法对硒肽进行表征, 采用酶偶联法进行GPx活力测定和酶动力学分析, 用噻唑蓝(MTT)比色法评价了硒肽的抗氧化效果. 结果表明, 该硒肽的存在形式为SecRGD的二聚体. 该硒肽具有GPx活力, 其催化谷胱甘肽(GSH)还原H2O2的GPx活力为5.54 U/μmol, 高于经典的GPx模拟物Ebselen. 稳态动力学分析结果表明, 该硒肽的催化机制为乒乓机制. 该硒肽具有分子量小, 易溶于水, 毒性低及可有效保护Vero细胞免受氧化损伤的优点, 具有作为抗氧化药物的应用前景.     

硒化壳聚糖抗K562细胞的实验研究

用MTT法和AO/EB荧光染色法观察了硒化壳聚糖对K 562肿瘤细胞株生长的影响。结果发现,硒化壳聚糖可有效地抑制K 562细胞生长,并呈量效、时效关系。经硒化壳聚糖作用后的细胞可明显出现核固缩、碎裂等凋亡形态改变。硒化壳聚糖可诱导K 562细胞凋亡,抑制其生长。     

硒化壳聚糖对K 562和K 562/ADM作用的比较研究

应用MTT法和细胞集落形成率法观察了硒化壳聚糖对K562和K562／ADM肿瘤细胞株生长的影响，结果发现硒化壳聚糖可有效地抑制两种K562细胞生长，呈量效关系。同等剂量的硒化壳聚糖对K562细胞的作用强度高于对K562／ADM细胞。硒化壳聚糖对耐药的细胞株可产生抑制作用，若与化疗药合用则可减慢甚至部分逆转临床上K562细胞耐药性的发生。     

硒化壳聚糖对NB4细胞作用的实验研究

应用MTT法、AO/EB荧光染色法和NBT还原法观察硒化壳聚糖对NB4肿瘤细胞株生长的影响。结果发现,硒化壳聚糖可有效地抑制NB4细胞生长,并呈量效、时效关系。经100 mg/L及以上剂量硒化壳聚糖作用后的细胞可明显出现核固缩、碎裂等凋亡形态改变。而经100 mg/L以下剂量硒化壳聚糖作用后,细胞可出现诱导分化成熟的表现。     

亮绿褪色光度法检测Fenton反应产生的羟自由基

建立了Fg^2＋-H2O2-亮绿的分析新体系,并用于羟自由基的检测。该法利用Fenton反应产生羟自由基,并加入亮绿显色剂,羟自由基使亮绿褪色。在632nm处测其△A值的变化,可间接测定羟自由基的产生量。探讨了最佳实验条件,并对苯甲酸与丹参等中草药的抗羟自由基氧化性能作了比较,以验证本法的有效性,得到较好的清除结果。该法稳定,可作为一种简便的筛选抗氧化剂的方法。     

基于果糖自氧化体系的抗氧化剂筛选方法

机体内果糖的自氧化过程中会产生多种自由基, 并最终转化为羟自由基, 苯甲酸钠可捕获羟自由基生成具有强荧光信号的羟基苯甲酸钠. 本文采用荧光光度法考察了影响果糖自氧化体系的各种因素, 建立了果糖自氧化产生羟自由基体系. 实验结果表明, 在果糖浓度为8.00 mmol/L, CuSO₄浓度为20.0 μmol/L, 苯甲酸钠浓度为24.0 mmol/L, pH=7.4, 温度为37℃及反应时间为24 h的条件下, 果糖自氧化体系最终可产生19.27 μmol/L的羟自由基. 抗氧化剂的存在可清除果糖自氧化过程中产生的自由基, 使最终生成的羟自由基的量减少, 从而导致生成的羟基苯甲酸钠减少, 荧光信号减弱, 由此建立了基于果糖自氧化体系的抗氧化剂筛选方法. 利用本评价体系考察了抗氧化剂盐酸小檗碱和阿魏酸的抗氧化能力, 实验结果表明, 中药标准品盐酸小檗碱和阿魏酸均能有效清除果糖自氧化体系产生的羟自由基, 其IC₅₀值分别为0.023和0.036 mmol/L.     

羟自由基在电极电解过程中的形成规律

对钛基二氧化铅电极电解过程中产生的^·OH进行了定量研究.结果表明,钛基二氧化铅电极在适当的电解条件下可以产生大量的自由基,碱性和高频条件下产生的自由基比酸性和低频条件下产生的自由基多.苯酚降解实验结果说明了其极强的氧化性能,在废水的深度处理方面有着极其广阔的应用前景.     

Identification of isomeric spin adducts of Leu-Tyr and Tyr-Leu free radicals using liquid chromatography-tandem mass spectrometry

Free radical species are generally short-lived due to their high reactivity and thus direct measurement and identification are often impossible. In this study we used a spin trap, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), to trap radical intermediates formed during the oxidation of isomeric dipeptides tyrosine-leucine (Tyr-Leu) and leucine-tyrosine (Leu-Tyr), induced by the hydroxyl radical. To investigate the influence of the amino acid position in the peptide chain on the oxidation and free radical generation, the spin adducts were characterized using LC-MS and MS(n) . We detected carbon and oxygen DMPO adducts and adducts bearing two DMPO, which were analyzed by MS(n) . Both alkoxyl and peroxyl radicals were identified. Radical intermediates were localized in Tyr during oxidation of Tyr-Leu, while radicals were identified in Leu and Tyr during oxidation of Leu-Tyr. DMPO adducts of acyl radical species formed from cleavage of the peptide backbone, promoted by the alkoxyl radical in α carbon of the N-terminal amino acid were observed. The results show that the amino acid position has an influence in the oxidation process, at least on small peptides, and that the α carbon of the N-terminal amino acid is more vulnerable to the attack of the electrophilic hydroxyl radical.     

Detection and characterization of hydroxyl radical adducts by mass spectrometry

The study of the influence of free radicals in the biological process depends primarily on the capacity to detect these reactive species. In this work we have studied the application of mass spectrometry to the identification of hydroxyl radical species. The detection and identification by collisional activation mass-analyzed ion kinetic energy spectrometry (CA-MIKES) of a spin adduct of DMPO with the hydroxyl radical [(DMPO + O) + H]+ (m/z 130) has demonstrated that mass spectrometry can be a powerful tool in the detection and identification of spin adducts of DMPO with hydroxyl radical species. We were also able to detect the capture of secondary free radicals using ethanol by detecting and identifying the corresponding adduct [(DMPO + ethanol) + H]+. Other spin adducts have also been detected and identified. We consider that the use of mass spectrometry is a relevant technique for the detection of free hydroxyl radicals, especially in complex mixtures, since mass spectrometry is able to discriminate these adducts in such situations. Moreover, using this approach, it was possible to identify new spin adducts.     

分子筛催化剂-亚硫酸盐体系降解水中对乙酰氨基苯酚

过渡金属诱导亚硫酸盐体系产生硫氧自由基是一种新型的高级氧化技术,比常规的高级氧化过程(基于羟基自由基等)有更优越的性能.我们对过渡金属离子活化过硫酸盐和亚硫酸盐的硫氧自由基生成链式反应过程、自由基用于氧化有机污染物的国内外研究现状进行了大致总结,并以SBA-15介孔分子筛为载体,采用浸渍法制备Co-SBA-15固相催化剂,将此固相催化剂用于活化亚硫酸钠,产生强氧化性自由基,以此来降解制药废水中典型污染物对乙酰氨基酚(APAP).通过正交试验,探讨了pH、温度、钴离子与亚硫酸根的比值(简称钴硫比)、气体含氧量等参数对污染物降解效果的影响.对催化剂进行表征,对体系中产生的自由基类型进行检测,结合实验结果和文献资料对降解产物进行了分析,探究了APAP的降解原理.     

间溴苯甲酸在Pt电极上的电氧化反应

采用循环伏安法和原位傅里叶变换红外(FTIR)光谱法研究了碱性介质中间溴苯甲酸(3-BBA)在Pt 电极上的电化学氧化反应. 3-BBA 在Pt 电极上有良好的电化学活性. 在较低的氧化电位作用下(1000 mV), 3-BBA自由基负离子先在Pt 电极上失去一个电子, 生成对应的自由基, 继而发生脱羧反应生成溴苯自由基阳离子和二氧化碳. 随着电位的正移, 吸附在电极表面的羟基自由基进攻溴苯阳离子, 发生脱溴反应, 生成苯酚. 随着电位的升高, 苯酚继续氧化生成苯二酚以及苯醌等, 部分苯环还可经开环得到马来酸和富马酸.     

Ruthenium(II)/(III) complexes of 4-hydroxy-pyridine-2,6-dicarboxylic acid with PPh3/AsPh3 as co-ligand: impact of oxidation state and co-ligands on anticancer activity in vitro

With the aim to develop more efficient, less toxic, target specific metal drugs and evaluate their anticancer properties in terms of oxidation state and co-ligand sphere, a sequence of Ru(II), Ru(III) complexes bearing 4-hydroxy-pyridine-2,6-dicarboxylic acid and PPh(3)/AsPh(3) were synthesized and structurally characterized. Biological studies such as DNA binding, antioxidant assays and cytotoxic activity were carried out and their anticancer activities were evaluated. Interactions of the complexes with calf thymus DNA revealed that the triphenylphosphine complexes could bind more strongly than the triphenylarsine complexes. The free radical scavenging ability, assessed by a series of in vitro antioxidant assays involving DPPH radical, hydroxyl radical, nitric oxide radical, superoxide anion radical, hydrogen peroxide and metal chelating assay, showed that the Ru(III) complexes possess excellent radical scavenging properties compared to those of Ru(II). Cytotoxicity studies using three cancer lines viz HeLa, HepG2, HEp-2 and a normal cell line NIH 3T3 showed that Ru(II) complexes exhibited substantial cytotoxic specificity towards cancer cells. Furthermore, the Ru(II) complexes were found to be superior to Ru(III) complexes in inhibiting the growth of cancer cells.     

Ultrasound effects on the antioxidative defense systems of Porphyridium cruentum

Ultrasound is a special physical stimulus that has a variety of biological effects. This study provides a first systemic investigation on the ultrasound-induced oxidation and protection actions of the antioxidant defense system in Porphyridium cruentum. The hydroxyl radical and superoxide anion radical scavenging ability of the cells and the electrolyte leakage of the cell membrane were examined. The change of glutathione and carotenoids produced with/without ultrasonic processing were measured; the enzyme activities of superoxide dismutase, catalase, and membrane bound ATPases (Na(+)/K(+)-ATPase, Ca(2+)/Mg(2+)-ATPase) were evaluated for either ultrasound-treated or untreated P. cruentum. The hydroxyl radical and superoxide anion radical scavenging ability of ultrasound-treated P. cruentum increase 49.8 and 76.0%, respectively, of which the electrolyte leakage and malonyldialdehyde accumulation are also found increased 48.6 and 48.0%, respectively, indicating a state of oxidative stress. A significant enhancement of the activities of superoxide dismutase by 53.5%, catalase, membrane bound ATPases (Na(+)/K(+)-ATPase, Ca(2+)/Mg(2+)-ATPase increased by 67.7 and 69.3%, respectively), and the increment of glutathione and carotenoids production are also observed. These results suggested that oxidative stress manifested by elevated reactive oxygen species levels and malonyldialdehyde contents might be resulted from the biophysical responses of P. cruentum to the physical stimuli, and most likely the enhanced antioxidation ability of the algal cells stimuli by ultrasonic comes from the enhancement of enzymatic and nonenzymatic preventive substances as observed in this work.     

An important role of intramolecular free radical reactions in antimalarial activity of artemisinin and its analogs

The kinetic schemes of intramolecular reactions of five analogs of artemisinin were built. The method of intersecting parabolas was used for the calculation of activation energies and rate constants of each elementary step of these schemes. The competition between monomolecular and bimolecular free radicals was taken into account. It was evidenced that the intramolecular oxidation of these compounds proceeds as a cascade of consecutive free radical reactions with the formation of hydroperoxide groups. The latter decompose via reactions with the Fe(II) complexes generating free radicals. Among the radicals formed, the hydroxyl radical was proved to play the key role. A correlation between the yield of hydroxyl radicals n(OH) and antimalarial activity of compounds (IC(50)) was observed. The dependence of index IC(50) on n(OH) is linear in the logarithmic coordinates: ln[IC(50)(Artemisinin)/IC(50)(Compound)] = -14.10 + 3.85 ×n(OH). The proposed scheme explains and demonstrates a strong dependence of the antimalarial effectiveness of a drug on the chemical structure.     

元宝枫黄酮提取物的抗氧化性能

制备了元宝枫黄酮的乙醇提取物.用电子自旋共振(ESR)法和光敏化合物微生物法检测了元宝枫黄酮提取物对芳香烃自由基、单线态氧自由基及混合氧自由基的抗氧化性能 结果显示,元宝枫黄酮提取物具有良好的综合抗氧化性.     

Laser-Induced Oxidation of Cholesterol Observed During MALDI-TOF Mass Spectrometry

Conditions for the detection of three odd-electron cholesterol oxidation peaks were determined and these peaks were shown to be artifacts of the matrix-assisted laser desorption time of flight (MALDI-TOF) process. Matrix choice, solvent, laser intensity and cholesterol concentration were systematically varied to characterize the conditions leading to the highest signals of the radical cation peaks, and it was found that initial cholesterol solution concentration and resultant density of solid cholesterol on the MALDI target were important parameters in determining signal intensities. It is proposed that hydroxyl radicals, generated as a result of laser irradiation of the employed 2, 5-dihydroxybenzoic acid (DHB) matrix, initiate cholesterol oxidation on the MALDI target. An attempt to induce the odd-electron oxidation peaks by means of adding an oxidizing agent succeeded using an acetonitrile solution of DHB, cholesterol, and cumene hydroperoxide. Moreover, addition of free radical scavengers reduced the abundances of some oxidation products under certain conditions. These results are consistent with the mechanism of oxidation proposed herein involving laser-induced hydroxyl radical production followed by attack on neutral cholesterol. Hydroxyl radical production upon irradiation of dithranol matrix may also be responsible for generation of the same radical peaks observed from cholesterol in dithranol by an analogous mechanism.