Asymmetric dimethyl-arginine metabolism in a murine model of cigarette smoke-mediated lung inflammation.

Abstract

There is increasing evidence that the endogenous nitric oxide synthase (NOS) inhibitor asymmetric dimethyl-arginine (ADMA) is involved in the pathogenesis of chronic lung diseases. One important regulator of this molecule is the ADMA-metabolizing enzyme dimethyl-arginine dimethyl-aminohydrolase (DDAH). The objective of this study was to determine whether perturbation of the ADMA-DDAH pathway contributes to lung inflammation following exposure to cigarette smoke (CS). For these studies, wild-type and DDAH transgenic mice were sham or CS-exposed. Serum ADMA levels were determined by mass spectrometry. ADMA content and DDAH expression were also visualized in mouse lung tissue by immunohistochemistry. DDAH expression was determined by real-time quantitative PCR (qPCR). Inflammation was assessed by H&E staining and analyses of total cell counts and fluid tumor necrosis factor (TNF)-α levels (using ELISA) in lung lavage fluid. NF-κB binding activity in mouse lung epithelial (LA-4) cells was assessed by a transcription factor-binding assay. The results indicated that the concentration of serum ADMA was increased following exposure to CS, and this corresponded with increased ADMA content in bronchial epithelial cells in lung tissue. Total lung DDAH expression was significantly decreased in lung tissue and cultured LA-4 cells following CS exposure. Addition of exogenous ADMA increased CSE-mediated NF-κB binding activity and TNFα production in LA-4 cells more than 2-fold compared to that in CSE-exposed controls. CS-mediated lung inflammation was significantly attenuated in DDAH transgenic mice compared to in wild-type controls. These findings demonstrated that lung ADMA metabolism was altered in mice following CS exposure and suggested that ADMA played a role in CS-mediated inflammation through increasing the presence of inflammatory mediators in lung epithelial cells.

• Authors: Elizabeth B Staab, Jason Weigel, Fengxia Xiao, Nandakumar Madayiputhiya, Todd A Wyatt, Sandra M Wells
• Journal: Journal of immunotoxicology (J Immunotoxicol) 2015 Jul-Sep Vol. 12 Issue 3 Pg. 273-82 ISSN: 1547-6901 [Electronic] England
• PMID: 25913572 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Tumor Necrosis Factor-alpha
• N,N-dimethylarginine
• Arginine
• Nitric Oxide Synthase
• Amidohydrolases
• dimethylargininase

Topics

• Amidohydrolases (genetics, metabolism)
• Animals
• Arginine (analogs & derivatives, metabolism)
• Cell Line
• Environmental Exposure (adverse effects)
• Epithelial Cells (metabolism)
• Humans
• Lung (immunology, metabolism, pathology)
• Mice
• Mice, Inbred C57BL
• Mice, Transgenic
• Nitric Oxide Synthase (antagonists & inhibitors)
• Pneumonia (chemically induced, metabolism)
• Smoking (adverse effects)
• Tumor Necrosis Factor-alpha (metabolism)