Dipyridamole and RA-642 Inhibit the Production of Superoxide Anion and Free Radical Damage to Rat Lens

Abstract: We studied the effects of dipyridamole and RA-642 on the production of superoxide anions and on oxygen radicals-induced lipid peroxidation in lens tissue homogenates from normal rats and rats given dipyridamole or RA-642 intraperitoneally. Superoxide production was evaluated by phenazine methosulphate (PMS)-induced nitroblue tetrazolium (NBT) reduction and lipid peroxidation by ferrous sulfate and ascorbic acid (FeAs)-induced malondialdehyde (MDA) production. Dipyridamole and RA-642 showed an inhibitory effect on both assays in the experiments with lens tissue homogenates from untreated or treated rats. The extent of inhibition, however, was significantly higher in pyrimidopyrimi-dinic-treated rats (range of inhibition at different times of incubation was 18% versus 23–57. for dipyridamole and 14% versus 73–80. for RA-642 in the assay of MDA production, and 10% versus 33–37. for dipyridamole and 2.5% versus 11–32. for RA-642 in the assay of NBT reduction). Concentrations of dipyridamole and RA-642 in lens tissue from treated animals could not be determined (< 0.001 μg/mg of tissue). Although both compounds inhibited lipid peroxidation induced by oxygen free radicals, the mechanism of action might include the role of adenosine as a mediator.     

Enhancement of rat and human phagocyte superoxide anion radical production by cadmium in vitro

The mechanism by which cadmium produced oxidizing effects in vivo is unknown. We show that cadmium enhances the production of superoxide anion radical (O-(2) .), a reactive oxygen species, in digitonin-stimulated phagocytes from man and rat. Cadmium concentrations ranging from 3.6 X 10(-2)M to 3.6 X 10(-4)M inhibited O-(2) . production in rat alveolar macrophages or human granulocytes. However, when activated in the presence of 3.6 X 10(-5)M cadmium, the production of O-(2) . was increased by a factor of 2.11 +/- 0.25 above control levels in human granulocytes and 3.6 +/- 0.62 above control levels in rat alveolar macrophages. This effect by levels of cadmium within the range of those occurring during in vivo toxicity might provide an explanation for the oxidizing effects of this metal ion.     

Beta-naphthylamine induces anion superoxide production in rat peritoneal macrophages.

Rat peritoneal macrophages were incubated in the presence of beta-naphthylamine (beta-NA), a well known carcinogenic agent, and some parameters of respiratory burst were studied. beta-NA induced a time- and dose-dependent stimulation of superoxide anion (O-2) production, and this enhancement was suppressed by the addition of superoxide dismutase enzyme. Also, no cooperative effect between beta-NA and phorbol 12-myristate 13-acetate was observed. Other observations were as follows: (i) the simultaneous presence of polymyxin B, and staurosporine inhibitors of protein kinase C, inhibited beta-NA-dependent O-2 production; (ii) NADPH-oxidase contained in postnuclear fraction from beta-NA-incubated macrophages showed a greater activity than control fractions; (iii) the stimulation of O-2 production elicited by beta-NA was several-fold enhanced in activated macrophages compared to resident cells. These data suggest that beta-NA produces the activation of NADPH-oxidase through protein kinase C.     

生脉溶液对超氧阴离子自由基作用的影响

目的:研究生脉溶液对超氧阴离子自由基(O-2)的影响.方法:建立能够产生超氧阴离子的二甲基亚砜(DMSO)碱性体系,采用电子顺磁共振(EPR)方法,测定加入不同浓度溶液后体系中超氧阴离子信号的强度,采用Vitc溶液作阳性对照.结果:加入浓度为20%,40%生脉溶液时,体系中超氧阴离子信号增强,且浓度为20%时与相同浓度对照组比较,增强显著(P<0.01);加入生脉溶液浓度增高至60%,80%及100%时,超氧阴离子信号强度逐渐减少并接近零,与相同浓度VC溶液对照组比较,减少显著(P<0.01).结论:生脉溶液对超氧阴离子自由基具有双向调节作用,较低浓度时增加超氧阴离子的产生,较高浓度时具有直接清除作用.     

超氧阴离子清除剂对家兔硝酸甘油耐受性影响的实验研究

目的探讨超氧阴离子自由基清除剂对家兔硝酸甘油耐受性的影响。方法取18只家兔按随机数字表分为硝酸甘油+维生素C组、硝酸甘油组、对照组各6只,剃净家兔胸部部分皮毛,对照组置0.9%氯化钠注射液(5mg)贴膜,硝酸甘油组置含5mg硝酸甘油贴膜,硝酸甘油+维生素C组置含5mg硝酸甘油贴膜,同时给予维生素C配10mL 0.9%氯化钠注射液灌胃。观察3组胸主动脉血管环对硝酸甘油的舒张反应、超氧阴离子水平和超氧化物歧化酶(SOD)活性。结果硝酸甘油组对不同浓度硝酸甘油血管舒张反应低于对照组、硝酸甘油+维生素C组(P<0.05);硝酸甘油组血管超氧阴离子含量高于对照组、硝酸甘油+维生素C组(P<0.05)。结论家兔持续经皮贴硝酸甘油72h可产生耐药性,超氧阴离子清除剂对硝酸甘油耐受性有明显改善作用。     

芦荟中芦荟苷清除超氧阴离子自由基的ESR研究

目的　从分子水平探讨芦荟保健美容的作用机制。方法　利用电子自旋共振 (ESR)法,以二甲基亚砜在碱性条件下产生超氧阴离子自由基(O-·2 )体系,检测芦荟中主要有效成分芦荟苷清除O-·2 的能力。结果　芦荟苷具有良好的清除超氧阴离子自由基(O-·2 )的作用,芦荟苷浓度为 0. 26mg/mL时,对O-·2 的清除率达 100%。结论　芦荟的保健美容作用可能与其主要有效成分芦荟苷具有清除超氧阴离子自由基的能力有关。     

物质对人中性粒细胞超氧阴离子生成的活化作用

神经肽P物质能激活多种参与炎症和免疫反应的细胞. 然而P物质直接刺激人粒细胞产生超氧阴离子(O^÷₂)需要较高的浓度(>10 μmol·L^-1). 本实验观察了低浓度P物质对人粒细胞超氧阴离子生成的活化作用. 结果表明, P物质在低浓度时(3 μmol·L^-1)能活化人粒细胞, 使?甲酰基-甲硫氨酰基-亮氨酰基苯丙氨酸(fMLP)刺激产生的超氧阴离子明显增多. P物质的这一活化作用呈剂量和时间依赖性. 在无细胞外钙的情况下，P物质没有这种活化作用. P物质活化fMLP引起的O^÷₂生成增多作用可被神经激肽(NK)-1受体阻断剂spentide (1 μmol·L^-1), 磷脂酶C抑制剂U-73122 (10 nmol·L^-1)以及Ca²⁺通道阻断剂尼卡地平 (1 μmol·L^-1)所抑制. 这些发现提示, P物质活化人粒细胞可能是经NK-1受体偶联的磷脂酰肌?醇代谢途径, 并且是细胞外钙依赖性的.     

Priming effect of substance Ponsuper oxideanionradical production inhumanneu trophils

神经肽Ｐ物质能激活多种参与炎症和免疫反应的细胞．然而Ｐ物质直接刺激人粒细胞产生超氧阴离子（Ｏ÷２）需要较高的浓度（＞１０μｍｏｌ·Ｌ－１）．本实验观察了低浓度Ｐ物质对人粒细胞超氧阴离子生成的活化作用．结果表明，Ｐ物质在低浓度时（３μｍｏｌ·Ｌ－１）能活化人粒细胞，使甲酰基－甲硫氨酰基－亮氨酰基苯丙氨酸（ｆＭＬＰ）刺激产生的超氧阴离子明显增多．Ｐ物质的这一活化作用呈剂量和时间依赖性．在无细胞外钙的情况下，Ｐ物质没有这种活化作用．Ｐ物质活化ｆＭＬＰ引起的Ｏ÷２生成增多作用可被神经激肽（ＮＫ）－１受体阻断剂ｓｐｅｎｔｉｄｅ（１μｍｏｌ·Ｌ－１），磷脂酶Ｃ抑制剂Ｕ－７３１２２（１０ｎｍｏｌ·Ｌ－１）以及Ｃａ２＋通道阻断剂尼卡地平（１μｍｏｌ·Ｌ－１）所抑制．这些发现提示，Ｐ物质活化人粒细胞可能是经ＮＫ－１受体偶联的磷脂酰肌醇代谢途径，并且是细胞外钙依赖性的     

大黄中蒽醌类成分清除氧自由基作用的研究

目的通过研究大黄中游离蒽醌类成分大黄酸、芦荟大黄素、大黄素、大黄素甲醚和大黄酚清除超氧阴离子自由基（O2.）的作用,从分子生物学的角度探讨大黄延缓衰老作用的可能机理。方法采用电子自旋共振技术（ESR）分别检测大黄中游离蒽醌类成分对O2.的清除作用。结果大黄中游离蒽醌类成分对O2.的清除作用,依次为大黄酸〉大黄酚〉大黄素〉大黄素甲醚〉芦荟大黄素。结论大黄所具有的延缓衰老的作用,可能与其所含的游离蒽醌类成分大黄酸、芦荟大黄素、大黄素、大黄素甲醚和大黄酚具有清除O2.的能力有关。     

Paraquat damage of rat liver mitochondria by superoxide production depends on extramitochondrial NADH.

Pure rat liver heavy mitochondrial fractions, in which the absence of significant microsomal contamination was confirmed by electron microscopy and by the lack of glucose-6-phosphatase activity, were used to demonstrate the effect of paraquat on mitochondrial ultrastructure in the presence of external NADH. Starved mitochondria (orthodox conformation) did not show O2 uptake or structural injury from either paraquat alone or NADH alone. Marked O2 uptake and structural breakage occurred only when paraquat and NADH were added in combination. These alterations were resistant to rotenone and malate plus glutamate or NADPH could not substitute for NADH. Paraquat was reduced anaerobically by the mitochondria in the presence of NADH, but not of NADPH. The addition of superoxide dismutase, ferricytochrome c or p-benzoquinone protected against the breakage of mitochondria caused by paraquat plus NADH. These results demonstrate that mitochondria may produce paraquat radicals in the presence of extramitochondrial NADH and thus generate superoxide anion radicals, resulting in structural injury to the mitochondria, by mechanisms that may involve the mitochondrial outer membrane rather than the electron transfer chain. These mitochondrial mechanisms in paraquat toxicity seemed to be more probable in vivo than are microsomal mechanisms; the latter are postulated to function in detoxication because phenobarbital diminished paraquat toxicity and SKF 525-A or cobaltous ions enhanced the toxicity.     

超氧阴离子对脑片谷氨酸释放的增强及依布硒的抑制

超氧阴离子对脑片谷氨酸释放的增强及依布硒的抑制易永杨祥良徐辉碧赵西龙１张亨山１秦钰慧１（华中理工大学化学系，武汉４３００７４；１中国预防医学科学院环境卫生监测研究所毒理室，北京１０００２１）兴奋性毒性是神经系统疾病中神经元损伤的一种重要病理机理，其特...     

Inhibition of superoxide anion production in macrophages by anti-inflammatory drugs.

The inhibition by anti-inflammatory drugs of the production of Superoxide anions (O₂^?) by isolated guinea pig macrophages was studied spectrophotometrically using NADH and lactate dehydrogenase. id₅₀ values were: 4 × 10^?7M (diclophenac sodium), 1 × 10^?6M (oxyphenbutazone), 1 × 10^?5M (indomethacin), 4 × 10^?5M (phenylbutazone), 7 × 10^?5M (mefenamic acid), 8 × 10^?5 M (flufenamic acid), 8 × 10^?5M (colchicine), 3 × 10^?4M (aspirin), 3 × 10^?4M (benzydamine), 10^?3M < (dexamethasone) and 10^?3M < (gold sodium thiomalate). They seemed to block the cell membrane-associated mechanism to produce Superoxide anions, since most of them did not abolish the generation of superoxide anions from the xanthine oxidase plus hypoxanthine system. Cytochalasin B, pyrogallol, ascorbate, NEM, l-epinephrine and chlorpromazine also inhibited, the production of Superoxide anion, but many non anti-inflammatory drugs were ineffective. This technique was evaluated as a screening method in vitro for nonsteroidal anti-inflammatory drugs.     

Inhibition of prostaglandin E2 and superoxide anion production in rat peritoneal macrophages by the calcium antagonists nifedipine and nisoldipine.

In order to clarify the possibility of an antiatherogenic action of the calcium antagonists nifedipine (CAS 21829-25-4) and nisoldipine (CAS 63675-72-9) the effect of nifedipine and nisoldipine on phorbol myristate acetate (PMA)- and calcium ionophore A23187-stimulated O2- and PGE2 production from macrophages was investigated. Nifedipine and nisoldipine inhibited dose-dependently PMA-stimulated O2- and PGE2 production, but not A23187-stimulated PGE2 production. The 50% inhibitory concentration (IC50) of nifedipine and nisoldipine for PMA-stimulated O2- production were 60 and 8 mumol/l, respectively, whereas those for A23187-stimulated O2- were 9.3 and 2.0 mumol/l. IC50 of nifedipine and nisoldipine for PMA-stimulated PGE2 production were 3.0 and 2.8 mumol/l, respectively. The release of [1-14C]-arachidonic acid from labeled macrophages stimulated with PMA was inhibited approximately by 39 to 43% in the presence of 20 mumol/l nifedipine and nisoldipine. The increase of (Ca2+)i in macrophages induced by A23187 could not be attenuated by nifedipine and nisoldipine, and (Ca2+)i level did not alter when stimulated with PMA. These results suggest that the inhibitory mechanism of nifedipine and nisoldipine for O2- production from the macrophages appears to directly inhibit the enzyme system of the NADPH-oxidase complex through the activation of protein kinase C, and that the inhibition of PMA-stimulated PGE2 production may be due to a decrease of phospholipase A2 through protein kinase C. On the basis of the inhibitory action on O2- and PGE2 production from the macrophages, a possible mechanism of antiatherogenic effect of calcium antagonists was discussed.     

Effects of phenolcarboxylic acids on superoxide anion and lipid peroxidation induced by superoxide anion.

The effects of phenolcarboxylic acids, caffeic acid, p-coumaric acid, and ferulic acid on the generation of superoxide anion and the production of lipid peroxide induced by superoxide anion were studied. Only ferulic acid anion among the phenolcarboxylic acids scavenged superoxide. Caffeic acid and ferulic acid inhibited lipid peroxidation induced by superoxide anion. These effects were comparable to those of superoxide dismutase or DL-alpha-tocopherol.     

Participation of superoxide free radical and Mn2+ in sulfite oxidation.

Sulfurous acid gas is a well-known air pollutant. The participation of superoxide ($\text{O}{}_2{}^{\mathrm{<mtext>?</mtext>}}$), a species of activated oxygen, in sulfite oxidation was investigated in relationship to this health hazard. The reduction of nitroblue tetrazolium (NBT) was markedly accelerated in the presence of the xanthine-xanthine oxidase system (X-XO), Mn²⁺ and SO₃^2?, but not by X-XO and Mn²⁺ or X-XO and SO₃^2? alone. This accelerated NBT reduction was partially suppressed by superoxide dismutase and was completely suppressed by allopurinol. Oxygen consumption was also markedly accelerated under to condition which caused the increase in NBT reduction. Lipid peroxidation of rat liver homogenate increased in the presence of X-XO, SO₃^2?, or both. This increased lipid peroxidation was definitely suppressed by Mn²⁺. From these observations, it is suggested that chain reactions involving sulfite oxidation are initiated by O₂^? generated from X-XO, and Mn²⁺ acts as a catalyst in the process.     

Effects of p38 MAPK Inhibitor on angiotensin II-dependent hypertension, organ damage, and superoxide anion production

Angiotensin II (Ang II) activates p38 mitogen-activated protein kinase (p38 MAPK) and increases reactive oxygen species (ROS), but the nature of the relationship in vivo is not fully understood. We assess the effect of SB239063AN, a highly selective, orally active, p38 MAPK inhibitor, on Ang II-dependent hypertension, target-organ damage and ROS production. Sprague-Dawley rats and MAPKAP kinase-2 knockout mice were infused with Ang II. Ang II infusion increased the levels of phosphorylated p38 MAPK in the heart and aorta. Production of superoxide anion and expression of NAD(P)H oxidase subunit gp91 in the aorta were increased 4- and 5-fold, respectively. In addition, Ang II infusion led to endothelial dysfunction, progressive and sustained hypertension, and cardiac hypertrophy. Treatment with SB239063AN (800 ppm in the diet) significantly attenuated the levels of phosphorylated p38 MAPK in the heart and aorta, reduced superoxide anion generation by 57% (P < 0.01), markedly suppressed gp91 mRNA expression, prevented endothelial dysfunction, and blunted both the hypertension and cardiac hypertrophy. Ang II-dependent hypertension was also significantly attenuated in MAPKAP kinase-2 knockout mice. The results suggest that Ang II induced hypertension, organ damage, and ROS production are possibly mediated by p38 MAPK and inhibition of p38 MAPK may offer a therapeutic approach for cardiovascular disease.     

Formation of the superoxide radical anion in decomposition of vicasol in aqueous solution

Translated from Khimiko-farmatsevticheskii Zhurnal, Vol. 23, No. 8, pp. 905–908, August, 1989.     

槲皮素-钼配合物对超氧阴离子和羟自由基的清除作用

目的 探讨槲皮素-钼配合物对超氧阴离子自由基(O2-.)和羟自由基(·OH)的清除作用.方法 分别用改良的邻苯三酚法和水杨酸法测定槲皮素一钼配合物对O2-.和·OH的清除作用,并与槲皮素比较.结果 配合物对O2-.和·OH均有显著的清除作用,对·OH的清除作用优于槲皮素(P＜0.05).结论 槲皮素-钼配合物具有较强的清除自由基作用.