Monocytes of patients with systemic sclerosis (scleroderma) spontaneously release in vitro increased amounts of superoxide anion
Monocytes of patients with systemic sclerosis (scleroderma) spontaneously release in vitro increased amounts of superoxide anion
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DOI：
10.1046/j.1523-1747.1999.00476.x
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发表时间：
1999-01-01
影响因子：
6.5
通讯作者：
Gabrielli, A
中科院分区：
文献类型：
--
作者：
Sambo, P;Jannino, L;Gabrielli, A
有研究表明，有毒氧自由基可能参与系统性硬化症（硬皮病）（SSc）的发病机制。由于导致自由基产生的细胞是未知的，我们的目的是（i）评估SSc患者的未经操作和佛波醇12-肉豆蔻酸酯13-乙酸酯刺激的单核细胞和多形核中性粒细胞产生超氧阴离子（O-2（. -））的能力;和（ii）调查O-2（. -）这些细胞产生的参与烟酰胺-腺嘌呤二核苷酸二磷酸氧化酶生化途径的激活。利用细胞色素c的超氧化物歧化酶还原反应来评价O-2（. -）的产生，SSc患者的未经操作的单核细胞产生更多的O-2（. -）与原发性雷诺现象患者和正常对照单核细胞相比（p = 0.0001），弥漫性皮肤受累和病程小于5年的患者释放量更高（p = 0.02）。该酶的胞质组分p47（phox）和p67（phox）都被转移到SSc患者的富集但未操作的单核细胞的质膜上。线粒体氧化酶的参与被排除在缺乏抑制O-2（. -）当单核细胞在鱼藤酮（一种线粒体氧化酶抑制剂）存在下孵育时，在用佛波醇12-肉豆蔻酸酯13-乙酸酯刺激后，SSc患者的单核细胞产生更多的O-2（.比对照组。在SSc患者中，未经治疗的多形核中性粒细胞产生的O-2（. -）比单核细胞（p = 0.0001），仅略高于原发性雷诺现象患者和正常对照组的多形核中性粒细胞（p = 0.03），总之，我们证明在硬皮病患者中，未经处理的和佛波醇12-肉豆蔻酸酯13-乙酸酯刺激的单核细胞体外释放增加的超氧阴离子的量，通过激活烟酰胺，腺嘌呤二核苷酸二磷酸氧化酶，因此，有助于在这种疾病中发现的氧化应激。
It has been suggested that toxic oxygen free radicals can be involved in the pathogenesis of systemic sclerosis (scleroderma) (SSc). Because the cells that contribute to the generation of free radicals are not known, our aim was (i) to evaluate the ability of unmanipulated and phorbol 12-myristate 13-acetate-stimulated monocytes and polymorphonucleate neutrophils of SSc patients to generate superoxide anion (O-2(.-)); and (ii) to investigate whether the O-2(.-) produced by these cells involved the activation of nicotinamide-adenine dinucleotide diphosphate oxidase biochemical pathway. Employing the superoxide dismutase-inhibitable reduction of cytochrome c to evaluate the generation of O-2(.-), unmanipulated monocytes of SSc patients generated more O-2(.-) than primary Raynaud's phenomenon patients and normal control monocytes (p = 0.0001), and the release was higher in patients with diffuse cutaneous involvement and 5 y or less disease duration (p = 0.02), The involvement of nicotinamide-adenine dinucleotide diphosphate oxidase in the enhanced O-2(.-) production was demonstrated by the finding that the cytosolic components of the enzyme, p47(phox) and p67(phox), Were both translocated to the plasma membrane of enriched but otherwise unmanipulated monocytes of SSc patients. The involvement of mitochondrial oxidases was excluded by the lack of inhibition of O-2(.-) production when monocytes were incubated in the presence of rotenone, a mitochondrial oxidase inhibitor. Upon stimulation with phorbol 12-myristate 13-acetate, monocytes of SSc patients produced more O-2(.-) than controls. In SSc patients untreated polymorphonucleate neutrophils generated significantly less O-2(.-) than monocytes (p = 0.0001) and only slightly more than polymorphonucleate neutrophils of primary Raynaud's phenomenon patients and normal controls (p = 0.03), In conclusion, we demonstrate that in patients with scleroderma, unmanipulated and phorbol 12-myristate 13-acetate-stimulated monocytes release in vitro increased amounts of superoxide anion through the activation of nicotinamide-adenine dinucleotide diphosphate oxidase and, thus, contribute to the oxidative stress found in this disease.