Involvement of endogenous hydrogen sulfide in cigarette smoke-induced changes in airway responsiveness and inflammation of rat lung

doi:10.1016/j.cyto.2010.12.006

Cytokine

Volume 53, Issue 3, March 2011, Pages 334-341

https://doi.org/10.1016/j.cyto.2010.12.006 Get rights and content

Abstract

Hydrogen sulfide (H₂S), recently considered the third endogenous gaseous transmitter, may have an important role in systemic inflammation. We investigated whether endogenous H₂S may be a crucial mediator in airway responsiveness and airway inflammation in a rat model of chronic exposure to cigarette smoke (CS). Rats randomly divided into control and CS-exposed groups were treated with or without sodium hydrosulfide (NaHS, donor of H₂S) or propargylglycine (PPG, inhibitor of cystathionine-γ-lyase [CSE], an H₂S-synthesizing enzyme) for 4-month exposure. Serum H₂S level and CSE protein expression in lung tissue were higher, by 2.04- and 2.33-fold, respectively, in CS-exposed rats than in controls (P < 0.05). Exogenous administration of NaHS to CS-exposed rats alleviated airway reactivity induced by acetylcholine (Ach) or potassium chloride (KCl) by 17.4% and 13.8%, respectively, decreased lung pathology score by 32.7%, inhibited IL-8 and TNF- α concentrations in lung tissue by 34.2% and 31.4%, respectively, as compared with CS-exposed rats (all P < 0.05). However, blocking endogenous CSE with PPG in CS-exposed rats increased airway reactivity induced by Ach or KCl, by 24.1% and 24.5%, respectively, and aggravated lung pathology score, by 44.8%, as compared with CS-exposed rats (all P < 0.01). Incubation in vitro with NaHS, 1–3 mmol/L, relaxed rat tracheal smooth muscle precontracted by Ach or KCl. However, the NaHS-induced relaxation was not blocked by glibenclamide (10⁻⁴ mol/L), L-NAME (10⁻⁴ mol/L), or ODQ (1 μmol/L) or denudation of epithelium. Endogenous H₂S may have a protective role of anti-inflammation and bronchodilation in chronic CS-induced pulmonary injury.

Introduction

Chronic obstructive pulmonary disease (COPD) is characterized by airflow obstruction due to chronic bronchitis or emphysema. Cigarette smoking is the major risk factor of COPD [1]. It induces an inflammatory response in the airways that might play a key role in the pathogenesis of COPD. Furthermore, multi-center clinical trials showed that current smokers with functional evidence of early COPD have airway hyper-responsiveness [2], [3]. Similarly, a dose-dependent effect of cigarette smoking on airway responsiveness was reported [4]. Airway hyper-responsiveness is an independent predictor of mortality in COPD patients [4]. However, the mechanisms of such hyper-responsiveness in COPD are less understood than are those in bronchial asthma.

The major effects of hydrogen sulfide (H₂S), long recognized as a toxic gas with a strong odor of rotten eggs in water pollution and industrial air pollution, are intoxication of the central nervous system and inhibition of the respiratory system [5]. Recently, H₂S was found endogenously generated in various mammalian tissues and may be a functional regulator in nervous and cardiovascular systems. H₂S is synthesized endogenously in various mammalian tissues by two pyridoxal-5′-phosphate-dependent enzymes responsible for metabolizing L-cysteine: cystathionine β-synthease (CBS) and cystathionine γ-lyase (CSE). The substrate of CBS and CSE, L-cysteine, can be derived from alimentary sources or can be liberated from endogenous proteins. It can also be synthesized endogenously from L-methionine through the trans-sulphuration pathway, with homocysteine being an intermediate in the process. Because of its endogenous metabolism and physiological functions, H₂S is well positioned in the novel family of endogenous gaseous transmitters and, in addition to nitric oxide (NO) and carbon monoxide (CO), might be the third endogenous signaling gasotransmitter [5]. Pulmonary tissue is rich in the H₂S synthase, cystathionine γ-lyase (CSE), and generates endogenous H₂S. Our previous study showed endogenous H₂S concentration increased in patients with stable COPD and decreased in patients with acute exacerbation of COPD; therefore, endogenous H₂S may be involved in the pathogenesis of airway inflammation and airflow obstruction in COPD [6], [7].

In this study, we investigated whether endogenous H₂S may be a crucial mediator in airway hyper-responsiveness and airway inflammation in a chronic cigarette smoke (CS)-exposed rat model.

Section snippets

Chronic cigarette smoke-exposed rat model

All animal care and experimental protocols were in compliance with the PR China Animal Management Rule (documentation 55, 2001, Ministry of Health of PR China) and the Third Hospital, Peking University Guide for the Care and Use of Laboratory Animals.

Male Sprague–Dawley rats weighing 200–250 g were supplied by the Animal Center, Health Science Center, Peking University, and randomly divided into six groups (each n = 6) for treatment: control, CS, sodium hydrosulfide (NaHS; an H₂S donor), NaHS + CS,

Chronic exposure to CS induced H₂S/CSE upregulation

As compared with controls, the NaHS-alone and NaHS + CS groups showed increased plasma levels of H₂S, by 2.70-fold and 3.29-fold, respectively, but the PPG-alone group showed decreased level, by 77.1% (all P < 0.01) (Fig. 1A). Exogenous administration of NaHS in NaHS + CS group increased the plasma level of H₂S, by 61.4%, whereas blockade of endogenous CSE with PPG in the PPG + CS group decreased the level, by 50.9%, as compared with the CS-alone group(all P < 0.01). Although the H₂S level of lung tissue

Discussion

The present study suggested that chronic exposure to CS can induce rat pulmonary injury and airway hyper-responsiveness. Endogenous H₂S generation was upregulated in the CS group. Administration of NaHS alleviated and PPG, inhibitor of CSE, aggravated lung injury and airway hyper-responsiveness in the CS group. The NaHS-induced relaxation in the rat tracheal was not blocked by inhibitors of K_ATP, NOS, sGC, or denudation of epithelium.

H₂S has been generally considered a toxic gas found in the

Acknowledgments

The work was supported by the National Natural Science Foundation of China (Grant No. 30871127), the Program for New Century Excellent Talents in University (Grant No. NCET-06-0003, 985-2-082-113) and the State Major Basic Research Development Program of the People’s Republic of China (2006CB503807).

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Wistar rats were divided into control and cigarettes smoke (CS) groups. CS group was daily subjected to cigarette smoke and treated by inhalation for 15 min/day with EUC (1 mg/mL) or vehicle during 30 days. After treatment, bronchoalveolar lavage (BAL) was collected to analyze the inflammatory profile, and tracheal rings were isolated for evaluation of the airway smooth muscle hyperresponsiveness. Lung function was analyzed in vivo.
The inflammatory profile was evaluated by optical microscopy performing total (Neubauer chamber) and differential leukocyte count (smear slides stained in H&E). The hyperresponsiveness was evaluated in tracheal rings contracted with potassium chloride (KCl) carbamoylcholine (CCh), or Barium chloride (BaCl₂) in presence or absence of nifedipine. The lung function (Newtonian resistance-R_N) was evaluated by bronco stimulation with methacholine (MCh).
BAL from CS group increased the influx of leukocyte, mainly neutrophils and macrophages compared to control group. EUC reduced by 71% this influx. The tracheal contractions induced by KCl, CCh or BaCl₂ were reduced by EUC in 59%, 42% and 26%, respectively. The last one was not different of nifedipine activity. Newtonian resistance (R_N) was also reduced in 37% by EUC compared to CS group.  
EUC reduces the hyperresponsiveness and the airway inflammatory profile, recovering the lung function.

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¹: These authors contributed equally to the development of the manuscript.

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Involvement of endogenous hydrogen sulfide in cigarette smoke-induced changes in airway responsiveness and inflammation of rat lung

Abstract

Introduction

Section snippets

Chronic cigarette smoke-exposed rat model

Chronic exposure to CS induced H2S/CSE upregulation

Discussion

Acknowledgments

Chest

Pulm Pharmacol Ther

Jpn J Pharmacol

Cytokine

Biochem Biophys Res Commun

J Sex Med

Curr Opin Pharmacol

Biochem Biophys Res Commun

Biochem Biophys Res Commun

J Pharmacol Sci

Biochem Biophys Res Commun

Biochem Biophys Res Commun

Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary

Am J Respir Crit Care Med

The lung health study: airway responsiveness to inhaled methacholine in smokers with mild to moderate airflow limitation

Am Rev Respir Dis

Chronic exposure to CS induced H₂S/CSE upregulation