Genetic targets of hydrogen sulfide in ventilator-induced lung injury--a microarray study.

Sashko Spassov,Dietmar Pfeifer,Stefan Ryter,Simone Faller,Matthias Hummel,Alexander Hoetzel,Karl Strosing,Utpal Sen

doi:10.1371/journal.pone.0102401

Abstract

Recently, we have shown that inhalation of hydrogen sulfide (H2S) protects against ventilator-induced lung injury (VILI). In the present study, we aimed to determine the underlying molecular mechanisms of H2S-dependent lung protection by analyzing gene expression profiles in mice. C57BL/6 mice were subjected to spontaneous breathing or mechanical ventilation in the absence or presence of H2S (80 parts per million). Gene expression profiles were determined by microarray, sqRT-PCR and Western Blot analyses. The association of Atf3 in protection against VILI was confirmed with a Vivo-Morpholino knockout model. Mechanical ventilation caused a significant lung inflammation and damage that was prevented in the presence of H2S. Mechanical ventilation favoured the expression of genes involved in inflammation, leukocyte activation and chemotaxis. In contrast, ventilation with H2S activated genes involved in extracellular matrix remodelling, angiogenesis, inhibition of apoptosis, and inflammation. Amongst others, H2S administration induced Atf3, an anti-inflammatory and anti-apoptotic regulator. Morpholino mediated reduction of Atf3 resulted in elevated lung injury despite the presence of H2S. In conclusion, lung protection by H2S during mechanical ventilation is associated with down-regulation of genes related to oxidative stress and inflammation and up-regulation of anti-apoptotic and anti-inflammatory genes. Here we show that Atf3 is clearly involved in H2S mediated protection.

Highlights

Mechanical ventilation can induce lung injury in the healthy lung or exacerbate pre-existing lung injury, both conditions which are referred to as ventilator-induced lung injury (VILI)
In a search for alternative therapeutic strategies, we recently found that inhalation of hydrogen sulfide (H2S) during mechanical ventilation prevents VILI in mice [4]
In addition to describing the genes differentially regulated in VILI [10,11,12,13,14,15,16], the present study focused on newly identified H2S target genes within several functional groups, including anti-inflammatory and anti-apoptotic pathways, regulation of extracellular matrix (ECM) remodelling and angiogenesis

Summary

Introduction

Mechanical ventilation can induce lung injury in the healthy lung or exacerbate pre-existing lung injury, both conditions which are referred to as ventilator-induced lung injury (VILI). VILI remains a major problem in critical care medicine [2], despite the implementation of low tidal volume ventilation that has reduced the rate of morbidity and mortality [3]. In a search for alternative therapeutic strategies, we recently found that inhalation of hydrogen sulfide (H2S) during mechanical ventilation prevents VILI in mice [4]. The protective properties of H2S have been increasingly investigated in various models of hemorrhagic shock, ischemia–reperfusion, oxidative stress, endotoxemia, and bacterial sepsis [5]. Gaseous H2S exerts anti-inflammatory effects by limiting cytokine release and neutrophil transmigration [4]. Application of H2S donors, such as sodium hydrosulfide (NaHS) and sodium sulfide (Na2S), exerts anti-inflammatory, antioxidant, and reducing effects that contribute to protection against VILI [8,9]

Objectives

Methods

Results

Discussion

Conclusion