Sambo P, Jannino L, Candela M, Salvi A, Donini M, Dusi S, Luchetti M M, Gabrielli A
Institute of Internal Medicine, Hematology and Clinical Immunology, University of Ancona, Italy.
J Invest Dermatol. 1999 Jan;112(1):78-84. doi: 10.1046/j.1523-1747.1999.00476.x.
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 (O2*-); and (ii) to investigate whether the O2*- 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 O2*-, unmanipulated monocytes of SSc patients generated more O2*- 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 02*- production was demonstrated by the finding that the cytosolic components of the enzyme, p47phox and p67phox, 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 O2*- 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 O2*- than controls. In SSc patients untreated polymorphonucleate neutrophils generated significantly less O2*- 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.
有人提出，毒性氧自由基可能参与系统性硬化症（硬皮病）（SSc）的发病机制。由于尚不清楚产生自由基的细胞，我们的目的是：（i）评估未经处理和经佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯刺激的SSc患者单核细胞和多形核中性粒细胞产生超氧阴离子（O2 * -）的能力；（ii）研究这些细胞产生的O2 * -是否涉及烟酰胺 - 腺嘌呤二核苷酸磷酸氧化酶生化途径的激活。利用超氧化物歧化酶可抑制的细胞色素c还原来评估O2 * -的产生，未经处理的SSc患者单核细胞产生的O2 * -比原发性雷诺现象患者和正常对照单核细胞更多（p = 0.0001），并且在弥漫性皮肤受累且病程为5年或更短的患者中释放量更高（p = 0.02）。烟酰胺 - 腺嘌呤二核苷酸磷酸氧化酶参与增强的O2 * -产生，这一发现表明该酶的胞质成分p47phox和p67phox都转位到了SSc患者富集但未经处理的单核细胞的质膜上。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，O2 * -产生缺乏抑制，这排除了线粒体氧化酶的参与。在用佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯刺激后，SSc患者的单核细胞产生的O2 * -比对照组更多。在SSc患者中，未经处理的多形核中性粒细胞产生的O2 * -明显少于单核细胞（p = 0.0001），仅略多于原发性雷诺现象患者和正常对照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，未经处理和经佛波酯12 - 肉豆蔻酸酯13 - 乙酸酯刺激的单核细胞通过激活烟酰胺 - 腺嘌呤二核苷酸磷酸氧化酶在体外释放增加量的超氧阴离子，从而导致该疾病中发现的氧化应激。
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