Effect of N-acetyl-L-cysteine on peroxynitrite and superoxide anion production of lung alveolar macrophages in systemic sclerosis.
Lung macrophages may play a relevant role in oxidative processes producing both superoxide anion (O(2)(-)) and NO. In this view, an antioxidant therapy can be useful in the treatment of systemic sclerosis (SSc) patients. N-Acetylcysteine (NAC) is able to expand natural antioxidant defenses by increasing intracellular gluthatione concentration and it has been proposed as an antioxidant therapy in respiratory distress syndromes. The aim of our study was to determine whether lung macrophages obtained from SSc patient bronchoalveolar lavage (BAL) express the inducible form of nitric oxide synthase (iNOS) and whether NAC can reduce the peroxynitrite (ONOO(-)) and O(2)(-) production of these cells. Alveolar macrophages were isolated from BAL of 32 patients and used for the immunocytochemical determination of iNOS, and the production of ONOO(-) and O(2)(-) was measured by fluorimetric or spectrophotometric methods, respectively. Lung macrophages obtained from SSc patients expressed a higher level of iNOS compared to healthy subject cells. NAC preincubation (5 x 10(-5)M, 24h) significantly reduced (-21%) the ONOO(-) production in formyl Met-Leu-Phe (fMLP)-activated cells and slightly reduced it under resting conditions, whereas NAC preincubation was unable to modify the release of O(2)(-) both in basal condition and in fMLP-stimulated cells. We conclude that since SSc lung macrophages express high levels of iNOS and produce a significant quantity of ONOO(-), NAC administration reduces ONOO(-) production and can be an useful treatment to alleviate SSc symptoms.
Failli
Paola
P
Department of Preclinical and Clinical Pharmacology, University of Florence, Viale Pieraccini 6, 50139, Firenze, Italy. failli@server1.pharm.unifi.it
Palmieri
Loredana
L
D'Alfonso
Caterina
C
Giovannelli
Lisa
L
Generini
Sergio
S
Rosso
Angela Del
AD
Pignone
Alberto
A
Stanflin
Nirvana
N
Orsi
Stefano
S
Zilletti
Lucilla
L
Matucci-Cerinic
Marco
M
D000111:Acetylcysteine / Q000494:pharmacology*; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000166:cytology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008168:Lung / Q000378:metabolism / Q000473:pathology; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D019001:Nitric Oxide Synthase / Q000096:biosynthesis; D052247:Nitric Oxide Synthase Type II; D030421:Peroxynitrous Acid / Q000032:analysis / Q000096:biosynthesis*; D012595:Scleroderma, Systemic / Q000378:metabolism* / Q000473:pathology; D013481:Superoxides / Q000032:analysis / Q000378:metabolism*
D013481:Superoxides; D030421:Peroxynitrous Acid; C496319:NOS2 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; D000111:Acetylcysteine