氧自由基的分析研究进展

本文对氧自由基的分析测定方法进行了综述和评价,主要包括电子自旋共振、高效液相色谱法、分光光度法、化学发光法、荧光和电化学等方法。     

盐酸羟胺自氧化法测定SOD活性的电化学方法研究

盐酸羟胺在碱性水溶液中发生自氧化反应,可产生O2-.,其自身被氧化为亚硝酸根离子,在酸性介质中SCN-催化亚硝酸根离子与二苯胺-4-磺酸反应生成具有电活性的亚硝基化合物在-0.60V处有一波形很好的二阶导数极谱峰。在一定条件下,极谱峰电流的大小与亚硝酸根离子的浓度呈线性关系。体系中加入SOD,使生成的亚硝酸根离子的量减小。由此建立了通过检测亚硝酸根离子的生成量来测定超氧化歧化酶活性的电化学方法,测定了酶活力曲线,并研究了常见的黄酮类化合物对O2-.的清除作用。     

芦荟超氧化物歧化酶的纯化与部分性质

芦荟粗提取液经丙酮沉淀、热处理、DEAE-琼脂糖柱层析和Sephacryl S-200凝胶过滤纯化后得到3种电泳纯的SOD组分,即SOD-Ⅰ、SOD-Ⅱ、SOD-Ⅲ.分子量分别为46000D,35000D,40000D;亚基分子量分别为23000D,17000D,20 000D,说明芦荟SOD三种组分均为二具体,其等电点分别为8.90,7.60,7.35。抑制剂敏感性实验和元素分析结果表明3种芦荟SOD组分均为Fe-SOD.     

芦荟超氧化物歧化酶(SOD)的分离纯化研究

采用硫酸铵分级沉淀、Sephadex G-100凝胶层析和DEAE-52柱层析,分离纯化了芦荟(A.vera L.)超氧化物歧化酶,纯化倍数为56倍,回收率为54%,比活力为643(U/mgpro),酶的最适pH值为8.2,最适温度在40~55℃,KCN和H2O2能抑制酶活性。实验表明:芦荟中的SOD属Cu、Zn-SOD。     

Icariin suppresses bone resorption activity of rabbit osteoclasts in vitro

The effect of icariin on the bone resorption activity of rabbit osteoclasts is assessed in vitro. Osteoclasts were isolated from Japanese white rabbits and cultured on plates with a sterilized bone slice in each well. After treatment with icariin at various concentrations, the bone resorption activity of osteoclasts was evaluated by examining pit areas, superoxide anion (·O2-) generation, size and number of actin rings and intracellular calcium concentration [Ca2 ]i. As revealed by these data, icariin elicited continuous decline of [Ca2 ]i, making actin ring constricted and ·O2- generation decreased. These events resulted in smaller and fewer pits which indicate suppressed bone resorption activity of rabbit osteoclasts by icariin.     

镉对草鱼的急性毒性效应及SOD的影响

以静水生物测试法研究了氯化镉对草鱼的急性毒性效应及其肝胰脏、肾脏和鳃组织的超氧化物歧化酶(SOD)的影响.结果表明:镉对草鱼的急性毒性作用时间为24,48,72和96 h的半数致死质量浓度(LC50)分别是45.58,34.81,28.63和24.05 mg.L-1,因此镉对草鱼的生态毒性为高毒.Cd2+低浓度时,肝胰脏和肾脏组织SOD活性均为"先降后升再降",而Cd2+为高浓度时SOD活性始终处于被显著抑制(p<0.05);鳃组织在低Cd2+污染时SOD活性为"先降后升",而高浓度的Cd2+污染时其SOD活性为"先降后升再降".但是草鱼鳃组织的SOD活性明显低于肝胰脏和肾脏的SOD活性.     

多巴胺的自氧化作用

研究了碱性溶液中多巴胺的自氧化作用，实验结果表明，多巴胺的自氧化可产生Ｏ＾－２，在一定范围内增加溶液的ｐＨ值和多巴胺的浓度均可增大Ｏ＾－２的产生速率。     

大球盖菇过氧化物酶及超氧化物歧化酶的研究

采用聚丙烯酰胺凝胶圆盘电泳法测定大球盖菇过氧化物酶和超氧化物歧化酶（SOD）同工酶活性,结果表明大球盖菇过氧化物酶有4种同工酶,比移分别是：0.13、0.18、0.25、0.32,其活性大小接近.SOD三种都有,比移分别为0.20、0.21、0.25,以Cu·Zn-SOD活性最大,Mn-SOD活性较小.Cu·Zn-SOD及Mn-SOD辅因子易于丢失,应用时应予以注意.     

Nitric oxide biosynthesis, nitric oxide synthase inhibitors and arginase competition for L-arginine utilization

Nitric oxide (NO) is a recently discovered mediator produced by mammalian cells. It plays a key role in neurotransmission, control of blood pressure, and cellular defense mechanisms. Nitric oxide synthases (NOSs) catalyze the oxidation of L-arginine to NO and L-citrulline. NOSs are unique enzymes in that they possess on the same polypeptidic chain a reductase domain and an oxygenase domain closely related to cytochrome P450s. NO and superoxide formation as well as NOS stability are finely regulated by Ca²⁺/calmodulin interactions, by the cofactor tetrahydrobiopterin, and by substrate availability. Strong interactions between the L-arginine-metabolizing enzymes are clearly demonstrated by competition between NOSs and arginases for L-arginine utilization, and by potent inhibition of arginase activity by N^ω-hydroxy-L-arginine, an intermediate in the L-arginine to NO pathway.     

活性氧对人体中与衰老相关物质的影响

研究了过硫酸按-TEMED体系产生的超氧负离子对健康人血液中几种生物活性物质的作用，结果表明，可以使细胞膜发生脂质过氧化作用，产生丙二醒（MDA），使生物膜造成损伤．同时使血液中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH－PX）活性均有所降低．通过对CAT纯品的研究揭示出在血液中经SOD，的作用产生毒性更强的·OH，它是使生物膜破坏及各种酶活性降低的主要原因．