Regulation of LPS-mediated inflammation in vivo and in vitro by the thiol antioxidant Nacystelyn

• Published in: American journal of physiology. Lung cellular and molecular physiology 2004-06, Vol.286 (6), p.L1319-L1327
• Main Authors: Antonicelli, Frank, Brown, David, Parmentier, Maryline, Drost, Ellen M, Hirani, Nik, Rahman, Irfan, Donaldson, Ken, MacNee, William
• Format: Article
• Language: English
• Subjects: Acetylcysteine - analogs & derivatives; Acetylcysteine - pharmacology; Animals; Antioxidants - pharmacology; Cells, Cultured; Chemotaxis - drug effects; Chemotaxis - immunology; Cycloheximide - pharmacology; Enzyme Inhibitors - pharmacology; Gene Expression - drug effects; Gene Expression - immunology; Humans; In Vitro Techniques; Interleukin-8 - genetics; Interleukin-8 - metabolism; Lipopolysaccharides; Lysine - analogs & derivatives; Lysine - pharmacology; Male; Monocytes - cytology; Monocytes - drug effects; Monocytes - immunology; Neutrophils - cytology; Neutrophils - immunology; Okadaic Acid - pharmacology; Pneumonia - drug therapy; Pneumonia - immunology; Protein Synthesis Inhibitors - pharmacology; Rats; Rats, Wistar; RNA, Messenger - analysis; Transcription Factors - metabolism
• ISSN: 1040-0605; 1522-1504
• DOI: 10.1152/ajplung.00329.2003

1 Edinburgh Lung and Environment Group Initiative/Colt Research Laboratories, Department of Medical & Radiological Sciences, University of Edinburgh Medical School; and 2 School of Life Sciences, Napier University, Edinburgh EH8 9AG, United Kingdom Submitted 15 September 2003 ; accepted in final form 2 February 2004 Increased levels of proinflammatory cytokines are present in bronchoalveolar lavage fluid in various lung diseases. Redox-sensitive transcription factors such as NF- B regulate gene transcription for these cytokines. We therefore studied the effect of a new thiol antioxidant compound, Nacystelyn (NAL), on IL-8 regulation in a human macrophage-derived cell line (THP-1). LPS (10 µg/ml) increased IL-8 release compared with control levels. This LPS activation was inhibited by coincubation with NAL (1 and 5 mM). Pretreatment with cycloheximide or okadaic acid, protein synthesis, and serine/threonine phosphatase inhibitors, respectively, did not modify inhibition of IL-8 release caused by NAL. NF- B and C/EBP DNA binding were increased after LPS treatment compared with control, an effect inhibited by cotreatment with NAL. Activator protein (AP)-1 DNA binding was unaffected. The enhanced neutrophil chemotaxis produced by conditioned media from LPS-treated cells was inhibited when cells were cotreated with NAL. The selectivity of NAL inhibition upon IL-8 expression was studied. LPS-treated THP-1 cells also had higher levels of TNF- , transforming growth factor (TGF)- 1 and -3, MIP-1 and - , and RANTES gene expression. However, only LPS-induced IL-8 and TGF- 1 expressions were inhibited by NAL. An anti-inflammatory effect of NAL was confirmed in vivo as shown by a reduction in LPS-induced neutrophil recruitment to the lungs following instillation of NAL into the lungs. Our studies demonstrate that NAL has anti-inflammatory properties in vitro and in vivo, may therefore have a therapeutic role in lung inflammation, and has the advantage over other antioxidant agents in that it may be administrated by inhalation. interleukin-8; lipopolysaccharide; THP-1 cells Address for reprint requests and other correspondence: W. MacNee, ELEGI/Colt Research Laboratories, Univ. of Edinburgh Medical School, Teviot Place, Edinburgh EH8 9AG, UK (E-mail: w.macnee{at}ed.ac.uk ).