Human SLPI inactivation after cigarette smoke exposure in a new in vivo model of pulmonary oxidative stress.
The role of oxidative stress in inactivating antiproteases is the object of debate. To address this question, we developed an in vivo model of pulmonary oxidative stress induced by cigarette smoke (CS) in mice. The major mouse trypsin inhibitor contrapsin is not sensitive to oxidation, and the mouse secretory leukoprotease inhibitor (SLPI) does not inhibit trypsin. Instead, human recombinant (hr) SLPI inhibits trypsin and is sensitive to oxidation. Thus we determined the effect of CS in vivo on hrSLPI antiproteolytic function in the airways of mice. CS caused a significant decrease in total antioxidant capacity in bronchoalveolar lavage fluid (BALF) and significant changes in oxidized glutathione, ascorbic acid, protein thiols, and 8-epi-PGF(2alpha). Intratracheal hrSLPI significantly increased BALF antitryptic activity. CS induced a 50% drop in the inhibitory activity of hrSLPI. Pretreatment with N-acetylcysteine prevented the CS-induced loss of hrSLPI activity, the decrease in antioxidant defenses, and the elevation of 8-epi-PGF-(2alpha). Thus an inactivation of hrSLPI was demonstrated in this model. This is a novel model for studying in vivo the effects of CS oxidative stress on human protease inhibitors with antitrypsin activity.
Cavarra
E
E
Department of Physiopathology and Experimental Medicine, University of Siena, I-53100 Siena, Italy.
Lucattelli
M
M
Gambelli
F
F
Bartalesi
B
B
Fineschi
S
S
Szarka
A
A
Giannerini
F
F
Martorana
P A
PA
Lungarella
G
G
D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D004781:Environmental Exposure; D016166:Free Radical Scavengers / Q000494:pharmacology; D006801:Humans; D008168:Lung / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018384:Oxidative Stress / Q000502:physiology*; D010947:Plants, Toxic; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins / Q000378:metabolism / Q000502:physiology*; D011994:Recombinant Proteins; D053481:Secretory Leukocyte Peptidase Inhibitor; D015843:Serpins; D012906:Smoke; D014026:Nicotiana; D014361:Trypsin Inhibitors / Q000032:analysis
D016166:Free Radical Scavengers; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D011994:Recombinant Proteins; C506814:SLPI protein, human; D053481:Secretory Leukocyte Peptidase Inhibitor; D015843:Serpins; C506815:Slpi protein, mouse; D012906:Smoke; D014361:Trypsin Inhibitors; C033691:contrapsin; D000111:Acetylcysteine