Original Contribution
Inhaled glutathione decreases PGE2 and increases lymphocytes in cystic fibrosis lungs

https://doi.org/10.1016/j.freeradbiomed.2005.03.032Get rights and content

Abstract

Reduced glutathione (GSH), a major antioxidant and modulator of cell proliferation, is decreased in the bronchoalveolar lavage fluid (BALF) of cystic fibrosis (CF) patients. We previously have shown that GSH inhalation in CF patients significantly increased GSH levels in BALF and improved lung function (M. Griese et al., 2004, Am. J. Respir. Crit. Care Med. 169, 822–828). GSH depletion in vitro enhances susceptibility to oxidative stress, increases inflammatory cytokine release, and impairs T cell responses. We therefore hypothesized that an increase in GSH in BALF reduces oxidative stress, decreases inflammation, and modulates T cell responses in lungs of CF patients. BALF from 17 CF patients (median FEV1 67% (43–105%) of predicted) was assessed before and after GSH inhalation for total protein, markers of oxidative stress (8-isoprostane, myeloperoxidase, and ascorbic and uric acid), pattern of protein oxidation, prostaglandin E2 (PGE2), and proinflammatory cytokines. BALF cells were differentiated using cytospin slides, and lymphocytes were further analyzed by flow cytometry. Inhalation of GSH decreased BALF levels of PGE2 and increased CD4+ and CD8+ lymphocytes in BALF significantly but had no effect on markers of oxidative stress. BALF lymphocytes correlated positively with lung function, whereas levels of PGE2 showed an inverse correlation. The patients with the greatest improvement in lung function after GSH treatment also had the largest decline in PGE2 levels. We conclude that GSH inhalation in CF patients increases lymphocytes and suppresses PGE2 in the bronchoalveolar space. Thus, GSH primarily affected the pulmonary immune response rather than the oxidative status in CF patients. The effect of GSH inhalation on PGE2 levels and lymphocytes in CF warrants further investigation.

Section snippets

Study design

The study population consisted of the previously reported 17 CF patients with mild to moderate lung disease (median FEV1 67% of predicted) (Table 1). Reduced GSH sodium salt (Biomedica Foscama, Ferentino, Italy) was reconstituted at concentrations of 200 or 300 mg/ml, resulting in a tonicity of 1301 or 1952 mosm/L. GSH was nebulized with a Pari LC Star nebulizer (Pari, Starnberg, Germany) and was inhaled three times daily. The total emitted volume was set to 1.5 ml by the AKITA inhalation

GSH administration

The aerosolized application of GSH for 2 weeks resulted in a three- to fourfold increase in BALF GSH levels with a mean pretreatment level of 2.9 μmol/L ± 0.7 SEM (substantially less than in normal individuals) and a mean posttreatment level of 12.4 μmol/L ± 4.2 SEM [22]. The GSH inhalation was well tolerated. Cough and an unpleasant odor from the GSH solution were reported by some patients.

Markers of oxidative stress

The delivery of substantial amounts of GSH into the lungs of CF patients did not change BALF

Discussion

We hypothesized that inhalation with GSH in CF patients would reduce oxidative stress, decrease inflammatory markers, and modulate lymphocyte response in BALF. The results show that the inhalation of GSH increased BALF lymphocytes, decreased BALF levels of PGE2, but did not affect the measured parameters of oxidative stress in BALF. No influence of GSH treatment on BALF levels of MPO, ascorbic acid, uric acid, and 8-isoprostane, as markers of oxidative stress, was found.

Furthermore, we found no

Acknowledgments

We thank Professor Luigi Inderst, Biomedica Foscama, Ferentino, Italy, and Omnia Pharma S.R.L., Italy, for providing the glutathione. This work was supported by grants from Mukoviszidose e.V., Bonn; CF-Selbsthilfe; Else-Kröner-Fresenius Stiftung; Friedrich-Baur-Stiftung; and a grant from the University and Science Program of the Ludwig-Maximilians-University (HWP).

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