地尔硫对冠心病的抗氧化作用

目的：观察地尔硫对冠心病病人的抗脂质过氧化及抗氧自由基作用。方法：冠心病病人３０例为治疗组（男性２１例，女性９例，年龄５８±ｓ６ａ），给予地尔硫３０ｍｇ，ｐｏ，ｔｉｄ，共４ｗｋ。健康人组３０例（男性１８例，女性１２例，年龄５８±９ａ）。结果：治疗组在给药前较健康人组的血清ｏｘＬＤＬ，ＬＰＯ，ＳＣＬ，ＬＣＬ值显著升高，Ｐ＜０．０１，ＧＳＨ－Ｐｘ显著降低，Ｐ＜０．０１，ＳＯＤ活力两者无明显差异，Ｐ＞０．０５。治疗组用药后与用药前相比ｏｘＬＤＬ，ＬＰＯ，ＳＣＬ，ＬＣＬ值显著下降，Ｐ＜０．０１或＜０．０５，ＧＳＨ－Ｐｘ显著升高，Ｐ＜０．０１，治疗组治疗后与健康人组相比，除ＬＣＬ外，其他指标皆无明显差异，Ｐ＞０．０５。结论：地尔硫是一良好的抗氧化剂。     

Direct measurement of phagosomal reactive oxygen by luminol-binding microspheres

A new method, utilizing microsphere-bound luminol, which makes possible the direct measurement of highly reactive oxygen within phagosomes, was studied. When Freund's complete adjuvant-elicited mouse peritoneal macrophages and luminol-binding microspheres were mixed, the microspheres were engulfed in macrophages and enclosed in phagosomes, where chemiluminescence (CL) was generated, showing the generation of highly reactive oxygen. The reactive oxygen could be quantitatively assayed by measuring the intensity of CL. The addition of cytochalasin B inhibited the CL. CL production by the thioglycollate-elicited macrophages was found to be only a ninth of that by Freund's complete adjuvant-elicited macrophages, though the phagocytic activities were almost equivalent in both cases.     

酶化学发光法测定何首乌中四羟基二苯乙烯葡萄糖甙的含量

报道一种新的何首乌中二苯乙烯甙含量的测定方法──酶化学发光法。该方法对二苯乙烯甙的检测限量为２．５×１０－７ｍｏｌ／Ｌ标准偏差小于２％，标准回收率在９６％～１０４％之间，线性范围为５．０×１０－７～１．０×１０－４ｍｏｌ／Ｌ。测定结果与紫外分光光度法基本一致。本法灵敏度较高，稳定性好，可用于中草药何首乌中二苯乙烯甙含量的测定。     

增强发光法测定过氧化物酶含量

本文介绍了一种应用增强发光法测定过氧化物酶含量的新方法。利用鲁米诺（Ｌｕ－ｍｉｎｏｌ）在过氧化物酶作用下与过氧化氢（Ｈ２Ｏ２）偶联的化学发光反应作基础，在增强发光剂一对碘酚的作用下，使其发光强度增强千倍，其检测灵敏度达到放射免疫测定水平已成为可能。     

基于纳米金催化鲁米诺-氯化汞化学发光新体系测定木犀草素

目的:基于纳米金对鲁米诺-氯化汞化学发光体系有良好的催化作用,建立测定木犀草素含量的高灵敏流动注射-化学发光分析法。方法:采用流动注射技术,实验研究了影响化学发光的各种因素;根据体系的化学发光光谱和紫外-可见吸收光谱结果,探讨了可能的化学发光机理。结果:0.1 mol·L-1氢氧化钠、4.0×10-4 mol·L-1鲁米诺、6.08×10-5 mol·L-1纳米金与1.0×10-3 mol·L-1 HgCl2组成最优的化学发光体系。在优化的实验条件下,本方法测定木犀草素的线性范围为2.0×10-9～1.0×10-7 g·mL-1(r=0.9929),检出限为1.5×10-9 g·mL-1,RSD为2.1%(C=1.0×10-8 g·mL-1,n=11)。纳米金作为电子转移的媒介,在鲁米诺-氯化汞反应过程中起催化作用;木犀草素和鲁米诺竞争与氯化汞发生反应,抑制了鲁米诺氧化成鲁米诺自由基,导致发光强度的降低。结论:利用本法成功地测定了血清与合成样品中木犀草素的含量。     

Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples

Introduction: The differentiation between extra‐ and intracellular production of reactive oxygen species (ROS) in whole blood was measured by luminol‐ and isoluminol‐enhanced chemiluminescence (CL). Methods: Azide (total CL inhibition), azide + horseradish peroxidase (HRP, restoring extracellular CL), superoxide dismutase + catalase (depleting extracellular ROS) and HRP (enhancing extracellular CL) were used to modulate luminol‐ and isoluminol‐enhanced CL (10^?6–10^?3 m luminophores) of 125× diluted whole blood which was activated by both calcium ionophore A23187 (Ca‐I) and opsonized zymosan particles (OZP) separately. Results: Both activators stimulated intra‐ and extracellular production of ROS. Luminol‐enhanced CL of Ca‐I‐activated samples detected the intracellular ROS, and with the addition of HRP detected the extracellular CL as well. CL enhanced with isoluminol in concentrations of 10^?4 m or less was mostly extracellular. There was a mixture of intra‐ and extracellular CL in OZP‐activated samples, probably because of the ingestion of luminophore molecules. Conclusion: Measurement of Ca‐I‐activated CL enhanced with 10^?4 m luminol is recommended for the detection of intracellular ROS. The addition of HRP leads to the detection of overall ROS production while the OZP‐activated system with its addition of HRP can only be used to detect overall ROS production. Ca‐I‐activated CL enhanced with 10^?4 m isoluminol and with addition of HRP is recommended for the detection of extracellular CL.     

Polymorphonuclear leukocyte chemiluminescence induced by Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus in serum and saliva

The ability of different strains of Actinobacillus actinomycetemcomitans (A.a .) and Haemophilus aphrophilus (H.a .) to trigger activation of an oxidative burst in human polymorphonuclear leukocytes (PMNL) was examined by measuring the luminol-amplified light emission - chemiluminescence (CL) - from these cells. Bacterial cells were incubated with PMNL from one healthy subject, in the presence of either active serum, heat-inactivated serum, saliva, or saliva and active serum. In the presence of active serum, all five H.a . strains and two out of five A.a . strains triggered a CL response. The CL induced in the presence of heat-inactivated serum was considerably less than that achieved with fresh serum. In the presence of only saliva, all strains induced considerably weaker CL responses than those induced in the presence of saliva with active serum. In the presence of serum, intracellular reactions appeared to be the main source of CL, while addition of saliva and active serum increased the extracellular CL. The results indicate that strain-dependent differences exist among A.a . strains in their ability to trigger the oxygen-dependent bactericidal mechanisms of human PNML. In contrast, the CL patterns of H.a . strains were equivalent. Various factors in the environment, such as activated complement and salivary compounds, affect the interaction of these species with neutrophils.     

Immunomodulatory effects of methoxylated flavonoids from two Chromolaena species: structure-activity relationships

Excessive generation of reactive oxygen species (ROS) by polymorphonuclear leukocytes (PMNL) is involved in the pathology of many inflammatory diseases. Compounds isolated from natural sources with antioxidant activity can be helpful to inhibit and/or modulate the oxidative damage associated with PMNL-derived ROS. The present study investigated the relationship between the chemical structure of five methoxylated flavonoids, isolated from Chromolaena hirsuta and Chromolaena squalida, and their inhibitory activity on ROS generation by opsonized zymosan-stimulated PMNL. The antioxidant efficacy of the studied flavonoids, assessed by luminol-dependent chemiluminescence, was dependent on the position and number of methoxy and hydroxy groups.     

Comparison of the effect of a mitochondrial uncoupler, 2,4-dinitrophenol and adrenaline on oxygen radical production in the isolated perfused rat liver

Using the isolated perfused rat liver, we examined the effect of stimulation of mitochondrial respiration by 2,4-dinitrophenol (2,4-DNP) and adrenaline on reactive oxygen species (ROS) production, liver damage and lipid peroxidation. ROS production was monitored by luminol- and lucigenin-enhanced chemiluminescence and oxygen uptake was measured simultaneously. Liver damage and lipid peroxidation were evaluated by measuring hepatic lactate dehydrogenase (LDH) and thiobarbituric acid reacting substances (TBARS) release. Tissue ROS level decreased and oxygen uptake increased soon after 2,4-DNP infusion. On termination of 2,4-DNP infusion, there was a sharp increase in lucigenin-enhanced chemiluminescence, which declined slowly, but luminol-enhanced chemiluminescence did not change prominently. Hepatic LDH and TBARS release increased gradually during 2,4-DNP infusion and were manifested by termination of the infusion. Allopurinol did not affect ROS production and TBARS release, but delayed increases in LDH release after termination of 2,4-DNP infusion. Adrenaline, which stimulates mitochondrial respiration without uncoupling caused similar but smaller ROS changes observed in 2,4-DNP. LDH and TBARS release were not affected significantly by adrenaline infusion. These results indicate that uncoupling of oxidative phosphorylation decreases ROS production and restoration of oxidative phosphorylation enhances ROS production and liver damage. Xanthine oxidase is unlikely to contribute to enhanced ROS production after termination of 2,4-DNP but has some protective effect during uncoupling.     

亚硒酸钠诱导白内障的机理研究

在pH8.0-8.5条件下,谷胱甘肽、氧或过氧化氢过量时,微量的10~(-4)mol·L~(-1)亚硒酸钠能使鲁米诺产生稳定的化学发光。发光强度依赖于酸度、谷胱甘肽浓度、氧及过氧化氢的浓度。超氧化物歧化酶(SOD)对鲁米诺发光有明显的抑制作用。SeO_3~(2-)在眼内催化产生活性氧可能是产生白内障的原因。