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Am. J. Respir. Crit. Care Med., Volume 163, Number 4, March 2001, 977-982

Lipopolysaccharide-induced Diaphragmatic Contractile Dysfunction and Sarcolemmal Injury in Mice Lacking the Neuronal Nitric Oxide Synthase

ALAIN S. COMTOIS, ESTHER BARREIRO, PAUL L. HUANG, ANDRÉ MARETTE, MYLÈNE PERRAULT, and SABAH N. A. HUSSAIN

Critical Care and Respiratory Divisions, Royal Victoria Hospital and Meakins-Christie Laboratories, McGill University, Montréal, Québec, Canada; Respiratory Division, Centre de Recherche du CHUM, Campus Notre-Dame, Université de Montréal, Montréal, Québec, Canada; Department of Physiology and Lipid Research Unit, Laval University Hospital Research Centre, St. Foy, Québec, Canada; and Cardiovascular Research Center, Massachusetts General Hospital, Boston, Massachusetts

In this study we evaluated the role of the neuronal nitric oxide synthase (nNOS) in lipopolysaccharide (LPS)-induced diaphragmatic contractile dysfunction and sarcolemmal injury. Wild-type (WT) mice or mice deficient in the nNOS gene (nNOS-/-) were injected with either saline (control) or Escherichia coli LPS (LPS groups) and sacrificed 12 h later. The diaphragm was then examined for NOS expression, NOS activity, and in-vitro contractility. We also assessed sarcolemmal injury in isolated muscle strips under resting condition and after 3 min of artificial stimulations. In WT mice, LPS injection reduced maximum force to about 75% of that of control animals and raised total NOS activity significantly due to the induction of the iNOS isoform. Although muscle fiber injury was minimal under resting condition, the percentage of injured fibers in control and LPS-injected mice approached 27% and 40% of total fibers, respectively, in response to artificial stimulation. By comparison, LPS injection in nNOS-/- mice elicited a worsening of muscle contractility (maximum force < 60% of control animals) but elicited degrees of sarcolemmal injury similar to those observed in the WT animals. In addition, muscle NOS activity and iNOS protein level in nNOS-/- mice injected with LPS reached about 10% and 60% of that of WT animals, respectively (p < 0.05 compared with WT animals). Protein level of endothelial NOS isoform in the diaphragm was not altered by LPS injection in either WT or nNOS-/- animals. We conclude that nNOS plays a protective role in attenuating the negative influence of sepsis on diaphragmatic contractility but is not involved in the pathogenesis of sepsis-induced sarcolemmal injury.




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