Inhibition of NF-κB Activation and Augmentation of IκBβ by Secretory Leukocyte Protease Inhibitor during Lung Inflammation

A Lentsch

J Jordan

B Czermak

K Diehl

E Younkin

V Sarma

P Ward

From the Department of Surgery,* University of Louisville School of Medicine, Louisville, Kentucky; the Departments of Pathology † and Surgery,‡ University of Michigan Medical School, Ann Arbor, Michigan; and the Department of Trauma Surgery,§ University of Freiburg, Freiburg/Breisgau, Germany

Accepted 1998 Sep 24.

Abstract

In earlier experiments, exogenous administration of secretory leukocyte protease inhibitor (SLPI) suppressed acute lung injury induced by deposition of IgG immune complexes. In the current studies we examined the mechanism of the protective effects of SLPI in this model. The presence of SLPI in the IgG immune complex-model of lung injury reduced the increase in extravascular leakage of ^{I-albumin, the intensity of up-regulation of lung vascular intercellular adhesion molecule-1, and the numbers of neutrophils accumulating in the lung. The presence of SLPI caused greatly reduced activation (ie, nuclear translocation) of the transcription nuclear factor-κB (NF-κB) in lung cells but did not suppress activation of lung mitogen-activated protein kinase. SLPI did not alter NF-κB activation in alveolar macrophages harvested 30 minutes after initiation of lung inflammation. In the presence of SLPI, content of tumor necrosis factor-α, CXC chemokines, and C5a in bronchoalveolar fluids was unaffected. In the inflamed lungs, inhibition of NF-κB activation by SLPI was associated with elevated levels of lung IκBβ (but not IκBα) protein in the absence of elevated mRNA for IκBβ. When instilled into normal lung, SLPI also caused similar changes (increases) in lung IκBβ. Finally, in the lung inflammatory model used, the presence of anti-SLPI caused accentuated activation of NF-κB. These data confirm the anti-inflammatory effect of SLPI in lung and point to a mechanism of anti-inflammatory effects of SLPI. SLPI appears to function as an endogenous regulator of lung inflammation.}

Abstract

During acute inflammatory injury of lung, neutrophils are recruited from the vascular space into interstitial and distal airway compartments. Secretory products of activated neutrophils, including oxidants and proteolytic enzymes, mediate lung parenchymal injury. Under normal circumstances, lung cells are protected from oxidant- and protease-mediated damage by a number of endogenous factors. Secretory leukocyte protease inhibitor (SLPI) represents an important component of the pulmonary anti-protease defense system. SLPI, a 12-kd protein that potently inhibits enzymes with serine protease activity, is constitutively expressed by a number of lung cell types including Clara cells of the bronchial epithelium and type II pneumocytes.^1-3 Recently, we have also shown that SLPI is expressed by pulmonary vascular endothelial cells in the inflamed lung.¹ It is known that inflammatory mediators induce SLPI expression in lung cells as well as in macrophages and neutrophils.^4-6 The protective properties of SLPI were originally thought to be due solely to its inhibitory effects on serine proteases because administration of SLPI reduced neutrophil elastase-induced emphysema.^7,8 However, other studies have suggested that SLPI increases the level of pulmonary glutathione, which may serve to reduce the extent of oxidant-mediated tissue injury.^9,10 SLPI also interferes with HIV entry into CD4^cells,¹¹ suggesting that its biological effects may extend beyond inhibition of serine proteases.

Acute lung injury induced by intrapulmonary deposition of IgG immune complexes in rats exhibits a pathophysiology similar to that observed during sepsis, ischemia/reperfusion, or trauma.^12-15 In this model, enhanced production of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) by activated lung macrophages causes up-regulation of the adhesion molecules, intercellular adhesion molecule-1 (ICAM-1), and E-selectin on the pulmonary endothelium.^16,17 Interactions of these vascular adhesion molecules with their respective ligands on blood neutrophils cause leukocyte adhesion to the endothelium and recruitment of neutrophils into the alveolar compartment. The ensuing lung injury is mediated by oxidants and proteases released by neutrophils and lung macrophages and is characterized by increased vascular permeability and alveolar hemorrhage.^18,19 During this inflammatory response, activation of the transcription factor, nuclear factor-κB (NF-κB), occurs in a time course similar to that for the production of TNF-α, IL-1β, and the expression of adhesion molecules in the pulmonary vasculature.²⁰

Administration of SLPI attenuates pulmonary recruitment of neutrophils and decreases lung injury induced by intrapulmonary deposition of IgG immune complexes.²¹ Blockade of endogenous SLPI results in an augmented inflammatory response with increased recruitment of neutrophils and lung injury (Gipson TS, Bless NM, Crouch LD, Shanley TP, Bleavins MR, Tefera W, McConnell PC, Mueller WT, Johnson KJ, Ward PA: Role of endogenous protease inhibitors in regulation of acute lung inflammatory injury. Submitted). However, the exact mechanisms by which SLPI exerts its anti-inflammatory effects in lung are unclear. Although SLPI inhibits serine proteases released by activated phagocytes, more recent studies have shown that SLPI interferes with the signal transduction pathway involved in the generation of matrix metalloproteinases.²² In the current studies, we sought to determine whether the protective effects of SLPI during lung inflammation might be caused by effects on other signal transduction mechanisms including NF-κB and mitogen-activated protein kinase (MAPK).

Values represent mean ± SEM with n = 4 for each group. SLPI (100 μg) was administered intratracheally with the anti-BSA.

Footnotes

Address reprint requests to Dr. Peter A. Ward, Department of Pathology, University of Michigan Medical School, 1301 Catherine Road, Ann Arbor, Michigan 48109-0602. E-mail: .ude.hcimu@drawp

Supported in part by the National Institutes of Health Grants GM-29587 and HL-31963 (P. A. Ward) and a grant from the American Lung Association (V. Sarma).

Footnotes

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