Department of Pharmacology and Pathophysiology, 1 Faculty of Pharmaceutical Sciences, and 2 Faculty of Biology, Utrecht University, 3508 TB Utrecht, The Netherlands; and 3 Department of Cardiovascular and Inflammation Pharmacology, Janssen Research Foundation, B-2340 Beerse, Belgium
![]() |
ABSTRACT |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The aim of this study was to determine the effects of glutathione (GSH) on trachea smooth muscle tension in view of previously reported interactions between GSH and nitric oxide (NO) (Gaston B. Biochim Biophys Acta 1411: 323-333, 1999; Kelm M. Biochim Biophys Acta 1411: 273-289, 1999; and Kharitonov VG, Sundquist AR, and Sharma VS. J Biol Chem 270: 28158-28164, 1995) and the high (millimolar) concentrations of GSH in trachea epithelium (Rahman I, Li XY, Donaldson K, Harrison DJ, and MacNee W. Am J Physiol Lung Cell Mol Physiol 269: L285-L292, 1995). GSH and other thiols (1.0-10 mM) dose dependently decreased the tension in isolated guinea pig tracheas. Relaxations by GSH were paralleled with sevenfold increased nitrite levels (P < 0.05) in the tracheal effluent, suggesting an interaction between GSH and NO. However, preincubation with a NO scavenger did not reduce the relaxations by GSH or its NO adduct, S-nitrosoglutathione (GSNO). Inhibition of guanylyl cyclase inhibited the relaxations induced by GSNO, but not by GSH. Blocking potassium channels, however, completely abolished the relaxing effects of GSH (P < 0.05). Preincubation of tracheas with GSH significantly (P < 0.05) suppressed hyperreactivity to histamine as caused by removal of tracheal epithelium. These data indicate that GSH plays a role in maintaining tracheal tone. The mechanism is probably an antioxidative action of GSH itself rather than an action of NO or GSNO.
nitrosothiols; epithelium; potassium channels; guanylyl cyclase
![]() |
INTRODUCTION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
AIRWAY HYPERRESPONSIVENESS is a key feature of several lung diseases. It is often associated with epithelial damage as a consequence of inflammatory processes (22). Damaged epithelium is impeded in its role of protecting the underlying smooth muscle against contractile stimuli. Moreover, the function of the epithelium as a source for relaxing factors that compensate for contractile stimuli will be impaired (15). One of those endogenous airway smooth muscle relaxants is nitric oxide (NO). NO is produced by a variety of cells and tissues in the respiratory tract, including the epithelial layer (4, 35). Under physiological conditions, genuine NO is very unstable and rapidly loses its biological activity by reacting almost instantaneously with oxygen, superoxide anion, and transition metals (16, 20, 23). Maintenance of an appropriate smooth muscle tone in the airways, therefore, requires continuous synthesis as well as stabilization of NO. Thiols are excellent candidates for the latter purpose. Under aerobic conditions, NO reacts with thiols to form nitrosothiols (RSNOs) via the nitrosylating intermediate dinitrogen trioxide (29). RSNOs are also produced by direct binding of nitrosonium ions to thiols (28). RSNOs can be regarded as stable pools of NO (18) and are themselves directly implicated in relaxing airway smooth muscle (24, 37). Whereas in principle, any thiol can bind NO, glutathione (GSH) is probably especially important in this respect. GSH is the major representative of the class of nonprotein thiols and plays a pivotal role in a variety of enzymatic and nonenzymatic reactions that protect tissues against oxidative stress (32). In view of the antioxidant role of GSH and widespread interactions between oxygen and tissues in the airways, it is not surprising that the airways are among the tissues containing the highest GSH concentrations in the body. Lung epithelial cells can be estimated to contain 10 mM GSH (39), and the epithelial lining fluid in the lungs contains 400 µM GSH, 100-fold higher than GSH levels in plasma (9).
In antioxidative reactions, GSH is converted into its oxidized form, glutathione disulfide (GSSG), that in its turn is enzymatically reduced into GSH to maintain a physiological redox balance. Under normal conditions, 95-99% of total GSH in the body is present in the reduced form (32). However, inflammatory diseases like asthma are associated with oxidative stress that places a large burden on the GSH pool (3, 11, 12). This may result in decreased levels of GSH available for NO stabilization and thus contribute to the development of airway hyperresponsiveness. Indeed, evidence was recently presented that an oxidative imbalance in the airways of asthmatics is reflected, among other parameters, by high levels of GSSG (27).
To address these issues, we tested whether addition of GSH to epithelium-denuded guinea pig tracheas increased NO levels as judged by a rise of nitrite levels in the perfusion buffer, and, if so, whether the increased nitrite levels showed a causal correlation with relaxation of tracheal smooth muscle. Because GSH-induced relaxations were indeed paralleled by a rise in nitrite levels in the buffer, we investigated whether guanylyl cyclase and potassium channels mediated these relaxations, since both guanylyl cyclase (6, 7, 26) and potassium channels (1, 25) are known play a role in NO-induced smooth muscle relaxation. Finally, the physiological relevance of alteration of tracheal tension by GSH was assessed by measuring whether perfusion of tracheas with GSH could moderate the hyperresponsiveness resulting from removal of their epithelium.
![]() |
METHODS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Animals and organ bath experiments. Male specific pathogen-free Dunkin Hartley guinea pigs weighing 350-400 g (Harlan Nederland, Horst, The Netherlands) were housed under controlled conditions. Water and commercial chow were allowed ad libitum. Guinea pigs were killed with an overdose of pentobarbital sodium (Nembutal; 0.6 g/kg ip body wt). Tracheas were dissected free of connective tissue and blood vessels, isolated, and divided into proximal and distal parts. Where indicated, the epithelial layer was removed from the tracheal segments as described earlier (14). Proper removal of the epithelium without causing damage to the underlying tissues was confirmed by light microscopy. Tracheas were mounted in perfused organ baths according to a modified method of Pavlovic et al. (36). The organ baths contained Krebs buffer (pH 7.4) of the following composition (mM): 118.1 NaCl, 4.7 KCl, 2.5 CaCl2, 1.2 MgSO4, 25.0 NaHCO3, 1.2 K2HPO4, and 8.3 glucose. The lumen of the trachea was perfused with Krebs solution independently from the outside by means of a peristaltic pump delivering a flow rate of 2 ml/min. The Krebs solution was continuously gassed with 5% CO2 in O2 and kept at 37°C. Two steel hooks were inserted through opposite sites of the tracheal wall with the smooth muscle between them. The lower hook was fixed to the bottom of the organ bath; the other hook was attached to an isometric force transducer (Harvard Bioscience, Kent, UK). Transducers were connected to an analog-to-digital converter, delivering digital signals to a computerized setup. The sampling frequency was 35 Hz. The setup allowed continuous sampling, online equilibrium detection, and real-time display of the responses on a computer screen.
The tracheal tension was set at an optimum counter weight of 4.0 or 2.0 g for thiol-induced relaxations and histamine-induced contractions, respectively. The use of different pre-tensions for assessing effects of relaxing and contractile agents is common practice in organ bath studies. The tissues were allowed to reach a stable tone for 60 min, during which the buffer was refreshed every 15 min. If necessary, tissues were allowed additional time to equilibrate without the buffer solution being changed.Thiol-induced tracheal relaxation. Epithelium-denuded tracheas were consecutively perfused with a range of concentrations (0.1-10 mM) of GSH, L-cysteine (Cys), or N-acetyl-L-cysteine (NAC). As a control, the nonthiol amino acid L-valine was tested at the same concentrations. In a separate set of experiments, GSH-induced relaxations were recorded in epithelium-denuded and in intact tracheal tubes, using the same range of concentrations as mentioned above.
Mechanisms of GSH-induced tracheal relaxations. For NO measurements, nitrite was assayed as a stable and representative breakdown product of NO formed enzymatically or NO released from RSNOs (28). Samples of 100 µl of tracheal effluent were collected just before or immediately after addition of GSH. The samples were injected into a purge vessel containing 2 ml of a 1% solution of sodium iodide in glacial acetic acid. The purge vessel was connected to a Sievers 270B NO analyzer (Boulder, CO). The sensitivity of the NO analyzer was 10 pmol/ml with a linearity of four log orders of magnitude. Calibrations were made according to the manufacturer's instructions with standard solutions of sodium nitrite (33).
In a separate set of experiments, the potential role of free NO in GSH-induced relaxations was investigated using the NO scavenger, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) (2). PTIO was added to the luminal buffer after the 60-min equilibrium period at a final concentration of 100 µM. Twenty minutes after adding PTIO, tracheas were relaxed by intraluminal addition of GSH, S-nitrosoglutathione (GSNO), or glyceryl trinitrate (GTN). The compounds were given in concentrations that evoke a submaximal response, i.e., 5.0 mM, 100 µM, and 10 µM for GSH, GSNO, and GTN, respectively. PTIO remained in the buffer during the relaxations. The inhibitor of soluble guanylyl cyclase (sGC) 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 50 µM) (21) or the nonselective potassium channel blocker tetraethylammonium (TEA; 20 mM) (31) was applied to both the mucosal and the serosal buffer. Controls received the vehicle of ODQ [DMSO; 0.25% (vol/vol) final concentration]. After 20 min of incubation, GSH (5 mM) or GSNO (100 µM) was added to the luminal buffer without removing TEA or ODQ, and relaxations were recorded. A 100-µl sample of the luminal perfusate of tracheas was taken before and after addition of GSH to be assayed for prostaglandin E2 (PGE2) content. PGE2 was quantified with an enzyme-linked immunoassay (Amersham, Roosendaal, The Netherlands) according to the manufacturer's instructions.Effect of GSH on tracheal responsiveness to histamine.
Epithelium-denuded tracheas were preincubated intraluminally with GSH
at a concentration of 1.0 mM or with saline (controls). After 40 min,
optimum tension (2.0 g) was readjusted mechanically, and contractions
were measured to increasing concentrations (108 to
10
3 M) of histamine in the inside buffer. GSH or saline
were left in the buffer during histamine-induced contractions.
Drugs. GSH, NAC, Cys, and L-valine were obtained from Sigma (St. Louis, MO). ODQ, GSNO, and PTIO were purchased from Alexis (Lausen, Switzerland). TEA was purchased from Merck (Darmstadt, Germany). GTN was obtained from Brocacef (Maarssen, The Netherlands).
Data analysis. Relaxations were determined as the percentage of the 4.0-g baseline tone that was set after completion of the equilibration period. Contractions were expressed as milligrams of tension on top of the 2.0-g baseline tone. Data are expressed as means ± SE. For most experiments, significance calculations were performed using the two-tailed Student's t-test. However, Wilcoxon's signed-rank test was used for assessing the statistics of the nitrite measurements, while a repeated measures analysis with a least significance differences post hoc test was used for analysis of the data pertaining to histamine reactivity. Differences were considered statistically significant if P < 0.05.
![]() |
RESULTS |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Thiol-induced tracheal relaxation.
GSH, as well as the sulfhydryl amino acids, Cys and NAC, relaxed
tracheas without epithelium dose dependently at concentrations of 1.0 mM and higher (Fig.
1A). Relaxations started
within seconds after administration of thiols (Fig. 1C shows
a representative tracing of a relaxant response to GSH; the profile of
the relaxations was similar for all thiols). The sensitivity to the
tested compounds increased in the order Cys GSH < NAC. When
administered at the highest concentration (10 mM), Cys, GSH, and NAC
reduced the initial 4.0-g baseline tension by ~20, 65, and 70%,
respectively. As expected, L-valine, which does not have a
thiol group, did not significantly alter baseline tension at any
concentration. GSH-induced relaxations were more extensive in denuded
tissues than in tissues with intact epithelium (Fig. 1B). At
10 mM, GSH reduced the initial 4.0-g baseline tension by 65.7 ± 6.5% in denuded tracheas and by 36.7 ± 5.9% in intact tracheal
tubes.
|
Mechanisms of GSH-induced tracheal relaxations.
Because NO is a well-known smooth muscle relaxant that can interact
with thiols, we measured whether perfusion with GSH increased nitrite
levels in the tracheal effluent. After the 60-min stabilization period,
i.e., just before addition of GSH, nitrite levels in the tracheal
effluent were 0.9 µM (95% confidence interval, 0.62-1.2), whereas immediately after addition of 10 mM GSH, levels increased significantly (P < 0.05) to 7.9 µM (95% confidence
interval 4.3-11.4; Fig. 2),
suggesting release of NO from the tissue by GSH.
|
|
|
Effect of GSH on tracheal responsiveness to histamine.
Perfusion of intact tracheas with histamine caused a moderate
concentration-dependent increase of smooth muscle tension, whereas contraction after removal of epithelium started at lower concentrations of histamine and was markedly stronger. This hyperresponsiveness upon
removal of epithelium has been reported earlier (34, 42). Preincubation with 1.0 mM GSH significantly attenuated the
hyperreactivity in tracheas without epithelium by 31.6 ± 9.46%
in terms of maximum response to histamine but did not change
responsiveness of tracheas with intact epithelium (Fig.
5).
|
![]() |
DISCUSSION |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
In this study, we have shown that GSH and other thiols reduced the smooth muscle tone of epithelium-denuded guinea pig tracheas in perfused organ baths. Relaxations up to 65-70% below baseline tension were induced by 10 mM GSH or NAC, the highest concentration tested (Fig. 1A). This concentration may be physiologically relevant because GSH is estimated to be present at this concentration in epithelial cells (39). Relaxations induced by Cys appeared less pronounced. Whether this indicates that the thiols need to be taken up in target cells to exert their relaxing effect is not clear. NAC, but not Cys, diffuses readily across the cell membrane. Uptake of GSH, however, has been shown to require an active transport mechanism that is present in epithelial cells but absent in many other cell types tested (10, 40, 43). In addition, the almost instantaneous relaxation induced by the thiols points to an extracellular rather than an intracellular mechanism. L-Valine failed to relax tracheas at any concentration, suggesting that induction of relaxation requires a sulfhydryl group and was not due to nonspecific effects such as osmolarity changes in the perfusion buffer. GSSG may be better than L-valine to control for these properties, but we obtained inconsistent results with GSSG. The compound had no effect on most tracheas tested (n = 6), but it relaxed some tracheas (n = 3) to the same extent as GSH. The latter observation is hard to explain. Because relatively high concentrations were required for relaxation, instantaneous reduction of GSSG to GSH by the latter tracheas seems unlikely.
Relaxations were more pronounced in epithelium-denuded tracheas than in intact tissues (Fig. 1B). The epithelium, therefore, possibly forms a physical barrier against relaxation by GSH on the underlying smooth muscle. Alternatively, intact epithelium already reduces tracheal tension by supplying the smooth muscle layer with GSH, so additional GSH can only have a limited effect.
Interestingly, GSH-induced relaxations in epithelium-denuded tissues were paralleled with a sevenfold rise in nitrite levels in the tracheal effluent (Fig. 2), suggesting release of NO by GSH (28). In view of the absence of the epithelium, the increase of nitrite levels has to be derived from sources other than the NO-rich epithelial cells. Sensory nerve endings in the trachea might have provided the NO and thus be the putative source of nitrite (5). It is doubtful, however, whether GSH caused relaxation by releasing genuine NO, in view of the effect of the free NO scavenger PTIO (2). This agent failed to inhibit relaxation by GSH, while it clearly inhibited relaxation by the genuine NO donor GTN (13) (Fig. 3). So, free NO, whether or not produced by nerve endings in the epithelium-denuded tracheas, is an unlikely mediator of the GSH-induced relaxation and nitrite formation. Alternatively, the GSH effects may involve nitrosylated proteins and other molecules in the subepithelial tissues. Those are likely to have been formed there before removal of the epithelium, because, once produced, NO can diffuse to neighboring cells and nitrosylate protein and nonprotein thiols via nitrosylating agents, like dinitrogen trioxide (29) and nitrosonium ions (28). GSH would then interact with tissue RSNOs to yield nitrite without the appearance of NO as an intermediate (41). It is also known that GSH forms GSNO in the presence of protein RSNOs (38). Although GSNO can cause tracheal relaxation, this molecule is also unlikely to be the mediator of the GSH-induced relaxations, since inhibition by ODQ of sGC, the primary target of NO and RSNOs (21), abolished the relaxing effect of exogenous GSNO, but not of GSH.
To see whether PGE2 would mediate the GSH-induced relaxations, this major relaxant prostanoid was measured in the perfusate of epithelium-denuded tracheas before and during the GSH-induced relaxation. PGE2 levels remained unaltered on addition of GSH to the organ bath buffer. These data show that PGE2 is not mediating GSH-induced relaxation.
In a further attempt to find a target of GSH that mediated the relaxations, the effects of the nonspecific potassium channel inhibitor TEA were investigated. TEA almost completely abolished trachea relaxation by GSH (Fig. 4), suggesting that GSH can activate particular potassium channels in this tissue. It is not unlikely that this is due to modification of sulfhydryl groups, since sulfhydryl reagents and other oxidizing compounds were reported to inactivate various potassium channels, while sulfhydryl reducing agents, like dithiothreitol and GSH, were shown to reverse inactivation or to cause their activation (8, 44). Further studies are needed to point out which potassium channel is involved in the observed relaxation and whether activation of that channel is regulated by sulfhydryl modification.
Interestingly, potassium channel inhibition also inhibited GSNO-induced trachea relaxation. Hence, in guinea pig trachea, GSNO apparently induces relaxation through cGMP-induced potassium channel activation. cGMP-dependent activation of calcium-activated potassium channels by RSNOs was reported earlier in rabbit coronary artery smooth muscle (19).
In addition to causing trachea relaxation, GSH also counteracted histamine-induced contraction in epithelium-denuded tracheas (Fig. 5). The more pronounced activity in epithelium-free tracheas is probably due to the same reasons pointed out above. The finding, however, is relevant because airway hyperresponsiveness is associated with sloughing of the epithelial layer (22).
Smooth muscle cell relaxation by GSH and other reduced thiols may represent a novel mechanism to maintain tracheal tone. Interestingly, this mechanism is probably not mediated by NO or GSNO, but possibly by an antioxidative action of GSH and other thiol compounds in the airways. Furthermore, the capacity of GSH to attenuate hyperreactivity in a model for damaged epithelium such as occurs in asthma (Fig. 5) suggests that replenishment of GSH in the airways might have therapeutic potential at physiological concentrations. Thiol replenishment in asthma has only been described once. Nebulization of GSH in mild asthmatics caused bronchoconstriction rather than a relief of symptoms (30), but this adverse effect may have been caused by the supraphysiological concentration of GSH (0.5 M), which was 500 times higher than the concentrations we used to dampen histamine-induced contractions. In a proper dose, replenishing the sulfhydryl content in the airways could be a potential therapy in diseases where excessive bronchoconstriction and oxidative stress are concomitant features.
![]() |
FOOTNOTES |
---|
Address for reprint requests and other correspondence: G. Folkerts, Dept. of Pharmacology and Pathophysiology, Faculty of Pharmaceutical Sciences, P.O. Box 80082, 3508 TB Utrecht, The Netherlands (E-mail: g.folkerts{at}pharm.uu.nl).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
April 12, 2002;10.1152/ajplung.00376.2001
Received 20 September 2001; accepted in final form 21 March 2002.
![]() |
REFERENCES |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
1.
Abderrahmane, A,
Salvail D,
Dumoulin M,
Garon J,
Cadieux A,
and
Rousseau E.
Direct activation of KCa channel in airway smooth muscle by nitric oxide: involvement of a nitrothiosylation mechanism?
Am J Respir Cell Mol Biol
19:
485-497,
1998
2.
Akaike, T,
Yoshida M,
Miyamoto Y,
Sato K,
Kohno M,
Sasamoto K,
Miyazaki K,
Ueda S,
and
Maeda H.
Antagonistic action of imidazolineoxyl N-oxides against endothelium-derived relaxing factor/NO through a radical reaction.
Biochemistry
32:
827-832,
1993[ISI][Medline].
3.
Antczak, A,
Kurmanowska Z,
Kasielski M,
and
Nowak D.
Inhaled glucocorticosteroids decrease hydrogen peroxide level in expired air condensate in asthmatic patients.
Respir Med
94:
416-421,
2000[ISI][Medline].
4.
Barnes, PJ,
and
Belvisi MG.
Nitric oxide and lung disease.
Thorax
48:
1034-1043,
1993[ISI][Medline].
5.
Belvisi, MG,
Miura M,
Stretton D,
and
Barnes PJ.
Endogenous vasoactive intestinal peptide and nitric oxide modulate cholinergic neurotransmission in guinea-pig trachea.
Eur J Pharmacol
231:
97-102,
1993[ISI][Medline].
6.
Belvisi, MG,
Ward JK,
Mitchell JA,
and
Barnes PJ.
Nitric oxide as a neurotransmitter in human airways.
Arch Int Pharmacodyn Ther
329:
97-110,
1995[ISI][Medline].
7.
Buga, GM,
Gold ME,
Wood KS,
Chaudhuri G,
and
Ignarro LJ.
Endothelium-derived nitric oxide relaxes nonvascular smooth muscle.
Eur J Pharmacol
161:
61-72,
1989[ISI][Medline].
8.
Cai, S,
and
Sauve R.
Effects of thiol-modifying agents on a K(Ca2+) channel of intermediate conductance in bovine aortic endothelial cells.
J Membr Biol
158:
147-158,
1997[ISI][Medline].
9.
Cantin, AM,
North SL,
Hubbard RC,
and
Crystal RG.
Normal alveolar epithelial lining fluid contains high levels of glutathione.
J Appl Physiol
63:
152-157,
1987
10.
Deneke, SM,
and
Fanburg BL.
Regulation of cellular glutathione.
Am J Physiol Lung Cell Mol Physiol
257:
L163-L173,
1989
11.
Dworski, R.
Oxidant stress in asthma.
Thorax
55, Suppl2:
S51-S53,
2000
12.
Dworski, R,
Murray JJ,
Roberts LJ, II,
Oates JA,
Morrow JD,
Fisher L,
and
Sheller JR.
Allergen-induced synthesis of F(2)-isoprostanes in atopic asthmatics. Evidence for oxidant stress.
Am J Respir Crit Care Med
160:
1947-1951,
1999
13.
Feelisch, M.
The use of nitric oxide donors in pharmacological studies.
Naunyn Schmiedebergs Arch Pharmacol
358:
113-122,
1998[ISI][Medline].
14.
Folkerts, G,
Engels F,
and
Nijkamp FP.
Endotoxin-induced hyperreactivity of the guinea-pig isolated trachea coincides with decreased prostaglandin E2 production by the epithelial layer.
Br J Pharmacol
96:
388-394,
1989[Abstract].
15.
Folkerts, G,
and
Nijkamp FP.
Airway epithelium: more than just a barrier!
Trends Pharmacol Sci
19:
334-341,
1998[ISI][Medline].
16.
Furchgott, RF,
Khan MT,
and
Jothianandan D.
Comparison of properties of nitric oxide and endothelium-derived relaxing factor: some cautionary findings.
In: Endothelium-Derived Relaxing Factors and Nitric Oxide, edited by Rubanyi GM,
and VanHoutte PM.. Basel: Karger, 1990.
17.
Garthwaite, J,
Southam E,
Boulton CL,
Nielsen EB,
Schmidt K,
and
Mayer B.
Potent and selective inhibition of nitric oxide-sensitive guanylyl cyclase by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one.
Mol Pharmacol
48:
184-188,
1995[Abstract].
18.
Gaston, B.
Nitric oxide and thiol groups.
Biochim Biophys Acta
1411:
323-333,
1999[ISI][Medline].
19.
George, MJ,
and
Shibata EF.
Regulation of calcium-activated potassium channels by S-nitrosothiol compounds and cyclic guanosine monophosphate in rabbit coronary artery myocytes.
J Investig Med
43:
451-458,
1995[ISI][Medline].
20.
Gryglewski, RJ,
Palmer RM,
and
Moncada S.
Superoxide anion is involved in the breakdown of endothelium-derived vascular relaxing factor.
Nature
320:
454-456,
1986[ISI][Medline].
21.
Hobbs, AJ.
Soluble guanylate cyclase: the forgotten sibling.
Trends Pharmacol Sci
18:
484-491,
1997[ISI][Medline].
22.
Hogg, JC.
The pathology of asthma.
Clin Chest Med
5:
567-571,
1984[ISI][Medline].
23.
Ignarro, LJ.
Biological actions and properties of endothelium-derived nitric oxide formed and released from artery and vein.
Circ Res
65:
1-21,
1989[ISI][Medline].
24.
Jansen, A,
Drazen J,
Osborne JA,
Brown R,
Loscalzo J,
and
Stamler JS.
The relaxant properties in guinea pig airways of S-nitrosothiols.
J Pharmacol Exp Ther
261:
154-160,
1992[Abstract].
25.
Johansson Rydberg, IG,
Andersson RG,
and
Grenegard M.
Effects of the nitric oxide-donor, GEA 3175, on guinea-pig airways.
Eur J Pharmacol
329:
175-180,
1997[ISI][Medline].
26.
Kannan, MS,
and
Johnson DE.
Modulation of nitric oxide-dependent relaxation of pig tracheal smooth muscle by inhibitors of guanylyl cyclase and calcium activated potassium channels.
Life Sci
56:
2229-2238,
1995[ISI][Medline].
27.
Kelly, FJ,
Mudway I,
Blomberg A,
Frew A,
and
Sandstrom T.
Altered lung antioxidant status in patients with mild asthma.
Lancet
354:
482-483,
1999[ISI][Medline].
28.
Kelm, M.
Nitric oxide metabolism and breakdown.
Biochim Biophys Acta
1411:
273-289,
1999[ISI][Medline].
29.
Kharitonov, VG,
Sundquist AR,
and
Sharma VS.
Kinetics of nitrosation of thiols by nitric oxide in the presence of oxygen.
J Biol Chem
270:
28158-28164,
1995
30.
Marrades, RM,
Roca J,
Barbera JA,
de-Jover L,
MacNee W,
and
Rodriguez-Roisin R.
Nebulized glutathione induces bronchoconstriction in patients with mild asthma.
Am J Respir Crit Care Med
156:
425-430,
1997
31.
McCann, JD,
and
Welsh MJ.
Calcium-activated potassium channels in canine airway smooth muscle.
J Physiol (Lond)
372:
113-127,
1986[Abstract].
32.
Meister, A,
and
Anderson ME.
Glutathione.
Annu Rev Biochem
52:
711-760,
1983[ISI][Medline].
33.
Menon, NK,
Pataricza J,
Binder T,
and
Bing RJ.
Reduction of biological effluents in purge and trap micro reaction vessels and detection of endothelium derived nitric oxide (edno) by chemiluminescence.
J Mol Cell Cardiol
23:
389-393,
1991[ISI][Medline].
34.
Munakata, M,
Huang I,
Mitzner W,
and
Menkes H.
Protective role of the epithelium in the guinea pig airway.
J Appl Physiol
66:
1547-1552,
1989
35.
Nijkamp, FP,
and
Folkerts G.
Nitric oxide and bronchial reactivity.
Clin Exp Allergy
24:
905-914,
1994[ISI][Medline].
36.
Pavlovic, D,
Fournier M,
Aubier M,
and
Pariente R.
Epithelial vs. serosal stimulation of tracheal muscle: role of epithelium.
J Appl Physiol
67:
2522-2526,
1989
37.
Perkins, WJ,
Pabelick C,
Warner DO,
and
Jones KA.
cGMP-independent mechanism of airway smooth muscle relaxation induced by S-nitrosoglutathione.
Am J Physiol Cell Physiol
275:
C468-C474,
1998
38.
Pietraforte, D,
Mallozzi C,
Scorza G,
and
Minetti M.
Role of thiols in the targeting of S-nitrosothiols to red blood cells.
Biochemistry
34:
7177-7185,
1995[ISI][Medline].
39.
Rahman, I,
Li XY,
Donaldson K,
Harrison DJ,
and
MacNee W.
Glutathione homeostasis in alveolar epithelial cells in vitro and lung in vivo under oxidative stress.
Am J Physiol Lung Cell Mol Physiol
269:
L285-L292,
1995
40.
Rahman, I,
and
MacNee W.
Lung glutathione and oxidative stress: implications in cigarette smoke-induced airway disease.
Am J Physiol Lung Cell Mol Physiol
277:
L1067-L1088,
1999
41.
Singh, SP,
Wishnok JS,
Keshive M,
Deen WM,
and
Tannenbaum SR.
The chemistry of the S-nitrosoglutathione/glutathione system.
Proc Natl Acad Sci USA
93:
14428-14433,
1996
42.
Tschirhart, E,
Frossard N,
Bertrand C,
and
Landry Y.
Arachidonic acid metabolites and airway epithelium-dependent relaxant factor.
J Pharmacol Exp Ther
243:
310-316,
1987[Abstract].
43.
Van Klaveren, RJ,
Demedts M,
and
Nemery B.
Cellular glutathione turnover in vitro, with emphasis on type II pneumocytes.
Eur Respir J
10:
1392-1400,
1997
44.
Wang, ZW,
Nara M,
Wang YX,
and
Kotlikoff MI.
Redox regulation of large conductance Ca2+-activated K+ channels in smooth muscle cells.
J Gen Physiol
110:
35-44,
1997
|
HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Visit Other APS Journals Online |