Am J Physiol Lung Cell Mol Physiol 283: L246–L255, 2002;

10.1152/ajplung.00491.2001.

invited review
Antioxidant responses to oxidant-mediated lung diseases

SUZY A. A. COMHAIR AND SERPIL C. ERZURUM

Departments of Pulmonary and Critical Care Medicine and Cancer Biology,
Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio 44195

Comhair, Suzy A. A., and Serpil C. Erzurum. Antioxidant re-

sponses to oxidant-mediated lung diseases. Am J Physiol Lung Cell Mol
Physiol 283: L246–L255, 2002; 10.1152/ajplung.00491.2001.—Reactive
oxygen species (ROS) and reactive nitrogen species (RNS) are generated
throughout the human body. Enzymatic and nonenzymatic antioxidants
detoxify ROS and RNS and minimize damage to biomolecules. An im-
balance between the production of ROS and RNS and antioxidant capac-
ity leads to a state of “oxidative stress” that contributes to the pathogen-
esis of a number of human diseases by damaging lipids, protein, and
DNA. In general, lung diseases are related to inflammatory processes
that generate increased ROS and RNS. The susceptibility of the lung to
oxidative injury depends largely on its ability to upregulate protective
ROS and RNS scavenging systems. Unfortunately, the primary intracel-
lular antioxidants are expressed at low levels in the human lung and are
not acutely induced when exposed to oxidative stresses such as cigarette
smoke and hyperoxia. However, the response of extracellular antioxidant
enzymes, the critical primary defense against exogenous oxidative
stress, increases rapidly and in proportion to oxidative stress. In this
paper, we review how antioxidants in the lung respond to oxidative
stress in several lung diseases and focus on the mechanisms that up-
regulate extracellular glutathione peroxidase.
redox; reactive oxygen species; reactive nitrogen species

OXYGEN IS ONE OF THE MOST abundant elements in our (RNS) such as nitric oxide, nitrite, and peroxynitrite
world, constituting 21% of the air we breathe (20, 122). (ONOO⫺) are both physiologically necessary and po-
It is essential for the oxidation of organic compounds, tentially destructive.
which is the process by which mammalian cells gener- Another oxygen-mediated mechanism of damage is
ate the energy needed to sustain life. However, oxygen inflammation, during which leukocytes, macrophages,
may also damage the lung. Inhaled ozone and nitric and mast cells release mediators that may cause bron-
oxide may induce toxic processes that impair lung choconstriction and edema as observed during an asth-
function (20, 38, 82, 119, 122). Under normal condi- matic reaction (15, 38, 64). Lung tissue can also be
tions, potentially toxic oxygen metabolites are gener- destroyed during reperfusion after an ischemic period
ated at a low level in lung cells by the transfer of a such as that produced by surgery (42, 94, 99, 104, 107,
single electron during aerobic metabolism (25, 33, 46). 134). All these mechanisms have one thing in common:
The resulting reactive oxygen species (ROS), which damage is at least partly mediated by oxidants and
include hydroxyl radicals, superoxide (O2⫺䡠), and hydro- nitrogen species.
gen peroxide (H2O2), play an integral role in the mod- To minimize oxidant damage to biological molecules,
ulation of several physiological functions but can also the human lung is endowed with an integrated antiox-
be destructive if produced in excessive amounts (31, 38, idant system of enzymatic and expendable soluble an-
82, 99, 104, 107). Similarly, reactive nitrogen species tioxidants. This system includes several antioxidant
defense mechanisms that detoxify reactive products or
convert them to products that are quenched by other
Address for reprint requests and other correspondence: S. A. A.
Comhair, Dept. of Pulmonary and Critical Care Medicine, Cleveland
antioxidants (47, 58). If the oxidant burden is suffi-
Clinic Foundation, 9500 Euclid Ave./NB4-107, Cleveland, OH 44195 ciently great, the reactive species may overwhelm or
(E-mail: [email protected]). inactivate the antioxidant system. The resulting excess
L246 1040-0605/02 $5.00 Copyright © 2002 the American Physiological Society http://www.ajplung.org

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 INVITED REVIEW L247

oxygen species can damage major cellular components, Much of the damage done by O2⫺䡠 and H2O2 in vivo is
including membrane lipids, protein, carbohydrates, due to their production of hydroxyl radicals (䡠OH) in a
and DNA. The pathophysiological consequences of this series of reactions catalyzed by traces of transition
injury are inflammation and widespread tissue dam- ions. One such example is the iron-catalyzed Haber-
age (46). Weiss reaction in which Fe3⫹ is reduced to Fe2⫹, fol-
lowed by the Fenton reaction in which the Fe2⫹ cata-
OXYGEN AND REACTIVE OXYGEN SPECIES lyzes the transformation of H2O2 into (䡠OH) (Eq. 4) (54).
More than 90% of all the oxygen we breathe under- O2⫺䡠 ⫹ Fe3⫹ 3 Fe2⫹ ⫹ O2 Haber-Weiss reaction
goes a concerted tetravalent reduction to produce wa- H2O2 ⫹ Fe 2⫹
3 Fe 3⫹ ⫺
⫹ OH ⫹ 䡠OH Fenton reaction
ter in a reaction catalyzed by cytochrome oxidase in the
mitochondrial electron transport chain. Oxygen (O2) (4)
can also be reduced via a nonenzymatic pathway An alternative pathway for 䡠OH formation in vivo
through four successive one-electron (e⫺) reductions (6, may involve MPO and EPO. Under physiological con-
34) (Eq. 1). centrations of halides, MPO produces hypochlorous
O 2 ⫹ 4H ⫹ ⫹ 4e ⫺ 3 2H 2O (1) acid (HOCl), and EPO produces hypobromous acid
(HOBr). Studies of 䡠OH with spin-trapping agents (41,
Cytochrome oxidase is the terminal electron acceptor 124) and chemical traps (57, 95) have demonstrated
in the respiratory chain and must donate its reducing that hypohalous acids can generate 䡠OH after reacting
equivalents to oxygen to allow continued electron with O2⫺䡠 (Eq. 5). 䡠OH can react with different molecules
transport. Otherwise, ATP production cannot continue. such as protein (16), DNA, and lipids (49).
Thus the major role for oxygen in all aerobic organisms O 2⫺䡠 ⫹ HOX 3 䡠OH ⫹ X ⫺ ⫹ O 2 (5)
is simply to act as a sink or dumping ground for
electrons (34). The tetravalent reduction of oxygen by RNS
the mitochondrial electron-transport chain is consid-
ered a relatively safe process. Nonetheless, the electron The discovery that nitric oxide (NO) is endogenously
carriers catalyze alternating one-electron oxidant-re- formed throughout the human body has led to intense
duction reactions, and they can react with oxygen to interest in the variety of roles this unique molecule
generate ROS such as O2⫺䡠 (47, 96, 97). Mitochondria plays in vivo. NO is involved in a wide variety of
are the major intracellular sites of O2⫺䡠 generation regulatory mechanisms. In addition, NO is also a cyto-
under physiological conditions (53). One other poten- toxic agent present in environmental pollutants and
tially major source for the generation of O2⫺䡠 is the cigarette smoke (109). NO is formed from the semies-
NADPH oxidase enzymatic system, which is found in sential amino acid L-arginine by the action of nitric
neutrophils, monocytes, macrophages, cytochrome oxide synthase (NOS; Fig. 1) (5, 90). Several forms of
P-450, monoamine oxidase, and lipooxygenase (4, 22, NOS have now been characterized, and several distinct
31, 34). O2⫺䡠 is also generated by other mechanisms NOS genes have been identified (73). The NOS are
such as molybdenum hydroxylase reactions (including classified as either constitutive or inducible (76, 89,
the xanthine, sulfite, and aldehyde oxidases) and ara-
chidonic acid metabolism.
O2⫺䡠 is relatively unstable, with a half-life of only
milliseconds. Because it is charged, it does not easily
cross cell membranes (6). O2⫺䡠 will react, however, with
proteins that contain transition metal prosthetic
groups, such as heme moieties or iron-sulfur clusters.
These reactions may damage amino acids or cause
protein/enzyme function to be lost (50, 138). Most of the
O2⫺䡠 generated in vivo undergoes a nonenzymatic or
superoxide dismutase (SOD)-catalyzed reaction, re-
sulting in the nonradical H2O2 (Eq. 2) (79). H2O2 can
also be directly produced by several oxidase enzymes,
including xanthine oxidase, monoamine, and amino
acid oxidase (24).
O 2⫺䡠 ⫹ O 2⫺䡠 ⫹ 2H ⫹ 3 H 2O 2 ⫹ O 2 (2)
H2O2 can be oxidized by eosinophil-specific peroxi-
dase (EPO) and neutrophil-specific peroxidase (MPO)
using halides (X⫺) as a cosubstrate to form the potent Fig. 1. Reactive nitrogen species (RNS) synthesis. NO, nitric oxide;
oxidant hypohalous acids (HOX) and other reactive NO3⫺, nitrate; NO2⫺ nitrite; HbO2, oxyhemoglobin; Hb3⫹, methemo-
halogenating species (Eq. 3) (44, 56, 70, 130). globin; ONOOH, peroxynitrous acid; MPO, myeloperoxidase; HOCl,
hypochlorous acid; SNO: S-nitrosothiol; NOS II, nitric oxide syn-
H2O2 ⫹ X⫺ ⫹ H⫹ 3 HOX ⫹ H2O X ⫽ Br⫺, Cl (3) thase II.

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L248 INVITED REVIEW

131). The constitutive forms (NOS I and NOS III) are and ONOOH) and (di)nitrosyl iron complex (52, 60, 66,
cytosolic and originally described and cloned from neu- 126, 132). Nitrosation of amines by these reactive ni-
ronal and endothelial cells, respectively. They are de- trogen intermediates has been implicated in the muta-
pendent on Ca2⫹ and calmodulin and release low genic properties of NO, presumably through nitrosa-
amounts of NO for short periods in response to receptor tive deamination of DNA bases (125). It is also of
and physical stimulation (89). The inducible form interest that SNO such as S-nitroso-L-glutathione
(NOS II) is independent of Ca2⫹. Once expressed, NOS (GSNO) may inhibit enzymes associated with the re-
II generates NO in large amounts for long periods sponse to oxidative stress in eukaryotic cells, including
(131). The biochemical effect of NO is largely defined by glutathione peroxidase (GPx), glutathione reductase
the concentration of NO. The paramagnetic NO mole- (7), glutathione-S-transferase (24), and ␥-glutamyl cys-
cule contains an odd number of electrons, which ex- teine synthase (55).
plains its highly reactive and radical nature (Fig. 1)
(65, 113). Autooxidation of NO with O2 results in the ANTIOXIDANTS
formation of nitrite (NO2⫺). However, at physiological
concentrations of NO and O2, this reaction may be too ROS and RNS play important physiological func-
slow to be important in vivo (8, 66). NO2⫺ is also a tions and yet they can also cause extensive damage.
substrate for hemeperoxidases such as MPO and EPO, The balance between physiological functions and dam-
which catalyze peroxidase-mediated oxidation and age is determined by the relative rates of formation
chlorination of biological targets (40, 41, 66, 124, 130, and the removal of ROS and RNS.
135, 136). Moreover, peroxidase-catalyzed oxidation of Normally, ROS and RNS are removed rapidly before
NO2⫺ results in the formation of a nitrogen dioxide they cause cellular dysfunction and eventual cell
radical (NO2 䡠) or related molecules. These substances death. All aerobic organisms use a series of primary
can contribute to the nitration of phenolic compounds antioxidant defenses to protect against oxidative dam-
such as tyrosine to form dimerized (dityrosine) and age. Furthermore, numerous repair enzymes remove
nitrated (3-nitrotyrosine) products, which are stable and/or repair damaged molecules. However, an antiox-
(40, 41, 66, 124, 130, 135, 136). idant cannot distinguish between radicals that play a
Although NO2⫺ is a major end product of NO, it does physiological role and those that cause damage (6). More-
not accumulate in vivo but is rapidly oxidized to nitrate over, some antioxidant compounds also have prooxidant
(NO3⫺) (91, 131). NO is also rapidly oxidized by oxyhe- actions (6). This section will review the enzymatic and
moglobin (HbO2), which results in the formation of nonenzymatic primary antioxidant defenses.
methemoglobin (Hb3⫹) and NO3⫺ (1, 14, 51). The oxida- Enzymatic antioxidants. SOD (EC 1.15.1.11) is an
tive metabolism of NO may also lead to the formation ubiquitous enzyme with an essential function in pro-
of carcinogenic nitrosoamines (72, 75) and the rapid tecting aerobic cells against oxidative stress (79). It
loss of NO’s smooth muscle relaxant activity (59, 117). catalyzes O2⫺䡠 radicals to H2O2. There are three forms
The rapid reaction of NO with free radicals (radical- of SOD. The copper-zinc SOD is located in the cytosol,
radical reaction) has emerged as one of the major the manganese SOD is primarily a mitochondrial en-
routes to the formation of RNS. At present, the best zyme, and extracellular SOD is usually found on the
understood of these reactions is the reaction with O2⫺䡠 outside of the plasma membrane (43).
to form ONOO⫺ (91), a strong oxidant (113). Although Catalase (EC 1.11.1.6) is a tetrameric hemoprotein
ONOO⫺ is relatively stable, it can be protonated to that undergoes alternate divalent oxidation and reduc-
yield peroxynitrous acid (ONOOH) (31), which then tion at its active site in the presence of H2O2 and
rapidly decomposes to NO3⫺ via the intermediate for- catalyzes the dismutation reaction (22, 36, 102). As a
mation of 䡠OH and NO2-like species (31). ONOOH is result, catalase has appreciable reductive activity for
very unstable, highly reactive, and capable of both small molecules such as H2O2 and methyl or ethyl
oxidizing and nitrating reactions. For instance, irre- hydroperoxide. It does not metabolize large molecular
versible ONOOH modifications include nitration of ar- peroxides such as lipid hydroperoxide products of lipid
omatic amino acids, lipids, or DNA bases (127). The peroxidation (129). Catalase is most effective in the
amino acid tyrosine appears to be particularly suscep- presence of high H2O2 concentrations. However, in the
tible to nitration, and the formation of free or protein- presence of low concentrations of either H2O2 or other
associated 3-nitrotyrosine has recently attracted inter- peroxides, the glutathione system plays a critical role
est as a potential biomarker for the generation of RNS (20).
in vivo (101, 125). The glutathione system is a central mechanism for
Reactions with thiol residues leading to the forma- reducing H2O2. It complements catalase as a reducing
tion of S-nitrosothiols (SNO) have been proposed as a system for H2O2 but exceeds catalase in its capacity to
mechanism whereby NO groups are transported and eliminate additional varieties of toxic peroxides (105).
targeted to specific effector sites, a potentially unique Other metabolized substrate species include large mol-
signaling mechanism induced by nitrosative stress (77, ecule lipid peroxides, formed by free radical attack on
85, 92). The exact mechanism by which S-nitrosation polyunsaturated lipid membranes and products of li-
occurs in vivo is still unclear, but it involves the for- pooxygenase-catalyzed reactions (58). The key enzyme
mation of NO-derived intermediates with the redox in the redox cycle responsible for the reduction of H2O2
equivalence of NO⫹ (the primary candidates are N2O3 is GPx. This reaction specifically requires reduced glu-
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 INVITED REVIEW L249

tathione (GSH) to serve as the electron donor. The min (transition metal binding), and cysteine and cys-
glutathione disulfide (GSSG) formed in the course of teamine (donators of sulfhydryl groups).
the reaction is subsequently reduced back to GSH by
glutathione reductase, which uses NADPH generated ANTIOXIDANTS IN THE LUNG
from the hexose monophosphate shunt system as an
Lungs are unique because they have a large epithe-
electron donor (37, 80). Healthy, nonstressed cells
lial surface area that is at risk for oxidant-mediated
maintain a high intracellular GSH:GSSG ratio to en-
attack. The tracheobronchial tree and the alveolar
sure the availability of GSH and thereby promote ac-
space are exposed to reactive oxidizing species in the
tive reduction of H2O2 through the glutathione system
form of inhaled airborne pollutants, tobacco smoke,
(37, 80). In a role unrelated to its role in the GSH
and products of inflammation. The lung, therefore,
system, free GSH can also function as a water-soluble
requires additional antioxidant resources to prevent
antioxidant by interacting directly with radical inter-
airway-borne oxidant injury (58). The major airways
mediates in nonenzymatic catalyzed reactions. Scav-
contain high-molecular-weight mucopolypeptide glyco-
enging of O2⫺䡠 by GSH leads to the formation of thiyl
proteins, which are synthesized by the epithelial cells
radicals (GS 䡠 ) and H2O2 via several steps, which is a
and glands that increase mucus production in the pres-
radical propagation reaction. This reaction leads to the
ence of inflammation (58). The lung contains intracel-
formation of GS 䡠 and H2O2 and can occur in physiolog-
lular antioxidant enzymes to maintain a normal redox
ically relevant concentrations. Hence, a substance that
state. The alveolar space can recruit additional antiox-
is generally accepted to be an antioxidant may possess
idant activity from the epithelial lining fluid (ELF).
prooxidant activity under certain conditions (6, 38).
This fluid contains large amounts of GSH (100-fold
Four GPx have been described, all selenium en-
higher than in plasma), 90% of which is in the reduced
zymes: 1) the classic cytosolic form, found in all cells
form (19, 27, 35, 114, 115). The ELF also contains
(9); 2) a membrane-associated GPx phospholipid H2O2
catalase, SOD, and GPx (19, 27, 114, 115). Additional
(39); 3) another cytoplasmic enzyme, gastrointestinal
antioxidants contained in ELF include ceruloplasmin,
GPx, which was first found in cells of the gastrointes-
transferrin, ascorbate, vitamin E, ferritin, other serum
tinal tract (23); and 4) an extracellular GPx (eGPx),
proteins, and small molecules such as bilirubin (58).
first identified as a distinct enzyme in human plasma
The multiplicity of the antioxidant systems available to
(137). All members of this family of enzymes can be
the lung and their overlapping specific activities sug-
oxidized by organic hydroperoxides, hydroperoxide, or
gest that to maintain normal pulmonary cellular func-
both, and can subsequently be reduced by glutathione
tion, it is critically important for the lung to adequately
(137). The existence of multiple forms of GPx is due to
control redox balance. Disequilibrium, either through
the expression of different genes (103). All GPx contain
increased oxidant stress or decreased antioxidant re-
a selenium atom in the active site in the form of
sources, can result in a series of pathophysiological
selenocysteine.
events in the lung that culminate in cellular death and
Nonenzymatic antioxidants. Cells use nonenzymatic
pulmonary dysfunction (58). A partial list of major
antioxidant compounds to react directly with oxidizing
lung diseases associated with oxidants is presented in
agents and disarm them. Such antioxidants are said to
Table 1.
be “scavengers”; their roles are unavoidably suicidal.
For example, vitamin E (␣-tocopherol) is a membrane- EXTRACELLULAR ANTIOXIDANT RESPONSE IN LUNGS
bound antioxidant that terminates the chain reaction EXPOSED TO OXIDATIVE STRESS
of lipid peroxidase by scavenging lipid peroxyl radicals
(LOO 䡠 ) (6, 34, 123). In this reaction, vitamin E becomes Normally, the homeostasis of cellular functions dur-
a radical, but it is much less reactive than LOO 䡠 (123). ing oxidative stress depends on the rapid induction of
However, at high concentrations, the radical form of protective antioxidant enzymes. Naturally occurring
vitamin E may function as a prooxidant (6). Vitamin C antioxidants exist to protect cells and tissue against
can also directly scavenge O2⫺䡠 and 䡠OH by forming the the continuous production of ROS/RNS during normal
semidehydroascorbate free radical that is subse- metabolism (58). Tissues and cells respond to mild
quently reduced by GSH (78). Vitamin C, however, is oxidative stress by increasing antioxidant defenses
usually not considered a major antioxidant because it (119). However, high levels of ROS/RNS may over-
also has prooxidant properties. It is probably the only whelm antioxidant defenses, resulting in oxidant-me-
cellular reducing agent other than O2⫺䡠 capable of con- diated injury or cell death (4, 20).
verting Fe3⫹ to Fe2⫹, which then reacts with H2O2 to Numerous studies have revealed that oxidant stress
form 䡠OH (106). Whether the prooxidant or antioxidant plays a crucial role in the initiation and progression of
properties of vitamin C prevail in any particular tissue a wide range of diseases and in the regulation of a
is determined by the extent of available iron stores; number of important biological processes. Pulmonary
iron overload favors excess oxidant generation (6, 106). diseases associated with oxidative stress include
Other nonenzymatic antioxidants include ␤-carotene asthma, hyperoxia, sarcoidosis, and chronic beryllium
(scavenger of O2⫺䡠 anions and peroxyl radicals), uric disease (CBD). ROS play a key role in the initial lung
acid (hydroxyl radical, O2⫺䡠, peroxyl radical scavenger), response to asbestos and silica that leads to interstitial
glucose (hydroxyl radical scavenger), bilirubin (LOO䡠 pulmonary fibrosis (87, 88). Interestingly, during the
scavenger), taurine (hypochlorous acid quencher), albu- development of pulmonary diseases, antioxidant re-
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L250 INVITED REVIEW

Table 1. Lung diseases associated with oxygen radicals

Disease Mechanism References

Emphysema Tissue injury by oxidants in cigarette smoke (21, 63)

Tissue injury by inflammatory cell oxidants ␣1PI inactivation by cigarette
smoke and inflammatory cells
Inflammatory cell release of oxidants ␣1PI oxidative inactivation by
Adult respiratory distress syndrome inflammation cells (26)
Hyperoxia Hyperoxia-mediated oxygen radical synthesis in cells (19)
Idiopathic pulmonary fibrosis Inflammatory cell oxidant release Glutathione deficiency (17, 18)
Asthma Inflammatory cell release of oxidants (35, 114)
Decrease in superoxide dismutase activity in bronchial epithelial cells
␣1PI, ␣-1-proteinase inhibitor.

sponses are different. For example, asbestosis and sar- tive stress occurring in individuals with asthma or
coidosis lead to an increase of SOD, whereas there are CBD and in those who have been exposed to exogenous
no changes found in silicosis or hyperoxic lung injury oxidants such as cigarette smoke (Fig. 2) (2, 28–30).
(27, 62, 71). In contrast, SOD activity is significantly The upregulation of eGPx occurs rather late after ex-
lower in patients with asthma and decreases further posure (after 24 h) (28), which may explain why eGPx
during an asthmatic exacerbation. was not induced after 12 h of hyperoxia. In support of
The glutathione system is altered in lung inflamma- this, levels of eGPx mRNA and protein increase only
tory conditions. For instance, GSH levels are elevated after 72 h of hyperoxia in a mouse model (67). Induc-
in the ELF of chronic smokers and in chronic beryllium tion of eGPx mRNA in bronchial epithelial cells is
disease, an immune-specific granulomatous inflamma- associated with elevated protein levels in ELF, sug-
tion (29). Levels of GSH in ELF decrease rapidly in gesting that the increase of eGPx occurs, in part, by
patients with mild asthma during an asthma exacer- bronchial epithelial cell synthesis and secretion (28).
bation (27). Similarly, GSH levels are decreased in However, alveolar macrophages can also express eGPx
ELF in idiopathic pulmonary fibrosis (17, 74), asbesto- (2, 30). It is not known whether other lung cells or
sis (11), acute respiratory distress syndrome (13), and inflammatory cells upregulate eGPx gene in response
in human immunodeficiency virus-positive patients to oxidative stress.
(12). Levels of glutathione modulate the T helper type Bronchial epithelial cells significantly increase eGPx
1 (Th1) vs. the Th2 immune response pattern (93). For mRNA expression in response to increased intracellu-
example, high levels of glutathione in patients with lar or extracellular ROS in vitro. Supplementation of
CBD may contribute to the development and/or main- GSH in cell culture to physiological levels potentiates
tenance of a chronic Th1 cell-mediated immune re-
sponse to beryllium, whereas the low GSH levels may
contribute to the development or maintenance of Th2
cell immune response in asthma.
Other enzymes of the glutathione system are also
influenced by oxidant stress. Some studies have shown
increased GPx activity in ELF of smokers compared
with nonsmokers (29, 104), whereas others have shown
decreased GPx in smokers (81). The difference in GPx
activity may be due to the difference in smoking his-
tory (19, 86). GPx activity is not altered in asthma but
is increased in lungs of CBD patients. Overall, the
antioxidant response is inconsistent across different
oxidant-mediated lung diseases.
EGPX AND ITS ROLE IN OXIDATIVE STRESS

Expression of eGPx. eGPx transcripts have been

found in epithelial cells with well-developed brush bor-
ders that contain lipids and alkaline phosphatase ac-
tivity, e.g., the human airways, intestine, and renal
tubules (2, 3, 68, 120). Alveolar macrophages are also
able to synthesize and secrete eGPx (2, 28, 30). The Fig. 2. Expression of extracellular glutathione peroxidase (eGPx)
GPx family is an important enzymatic component of mRNA in human airway epithelial cells (means ⫾ SE). No significant
the mechanisms for detoxifying ROS in the lung and difference in the expression of eGPx mRNA was found in cells from
individuals with asthma, chronic beryllium disease (CBD), and
may play a significant role in preventing pulmonary smoke exposure (P ⬎ 0.05), but in all 3 groups, levels were signifi-
oxidant stress. eGPx gene expression is upregulated in cantly higher than in controls (P ⬍ 0.05). GAPDH, glyceraldehyde-
bronchial epithelial cells and ELF as a result of oxida- 3-phosphate dehydrogenase.

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 INVITED REVIEW L251

the induction of eGPx mRNA in response to ROS (28).

This effect is not reproduced by N-acetylcysteine, sug-
gesting that other effects of thiol groups are necessary
to achieve the synergistic effect on induction of the
eGPx gene (28). Although GSH is usually considered
an antioxidant, it can also act as an oxidant when
present at physiological levels (6). GSH may partici-
pate in oxidizing processes and/or accelerate the gen-
eration of ROS, specifically O2⫺䡠 (6, 84, 121, 133). Pre-
vious reports have shown that GPx can function as an
ONOO⫺ reductase and thereby prevent nitration reac-
tions caused by RNS (45). On the other hand, NO
donors (S-nitroso-N-acetyl-D,L, penicillamine/GSNO)
induce eGPx gene expression in a time- and dose-
dependent matter.
Transcriptional regulation. The regulation of genes
in response to oxidative stress occurs via transcription
and/or stabilization of mRNA. Studies support that the Fig. 4. Detoxification of ROS and RNS by eGPx in asthmatic indi-
ROS regulation of the eGPx gene expression occurs at viduals. a: Superoxide is detoxified through eGPx to H2O; b: in the
a transcriptional level (28). In general, ROS and RNS presence of low levels of superoxide dismutase, NO and superoxide
(O2⫺䡠) combine to form ONOOH; c: peroxynitrite is able to nitrate
regulate the expression of numerous genes via signal- tyrosine residues; d: nitrosation of reduced glutathione (GSH) by
ing mechanisms. Redox-sensitive transcription factors peroxynitrite leads to S-nitroso-L-glutathione (GSNO); e: detoxifica-
such as signal transducers and activators of transcrip- tion and liberation of NO through eGPx. GSSG, glutathione disul-
tion (STAT), nuclear factor-␬B, and transcription fac- fide.
tor activator protein-1 (AP-1) are activated in epi-
thelial cells and inflammatory cells during oxidative
STATs by oxidative stress is inhibited by antioxidants.
stress (98, 100). Although the STAT family of tran-
Several ROS-induced target genes have known STAT
scription factors is activated by many cytokines and
binding sites in their promoters. These include genes
growth factors, it can also be activated by oxidative
involved in antioxidant defense (111) such as SOD1
stress such as H2O2 (108, 112, 116). The activation of
(10) and genes involved in cell growth regulation such
as c-fos (128).
Studies from several laboratories have demonstrated
that oxidant stress such as cigarette smoke, treatment
with H2O2, depletion of intracellular GSH, or an in-
crease in the GSH:GSSG ratio stimulates AP-1 activa-
tion and binding (83, 99). AP-1 regulates many of the
inflammatory and immune genes in oxidant-mediated
diseases (69, 110, 118). AP-1 is a protein dimer com-
posed of the Jun and Fos gene products (100). These

Fig. 3. Proposed scheme for eGPx gene induction by oxidative stress.

Reactive oxygen species (ROS) and RNS cross the cell membrane,
which together with inflammatory mediators activate different tran-
scription factors, such as nuclear factor (NF)-␬B and activator pro-
tein-1 (AP-1). The 5⬘-flanking region of the eGPx gene necessary for
transcriptional activation shows that the inducible promoter ele- Fig. 5. Nitrotyrosine staining of asthmatic bronchial mucosa. Cyto-
ment contains the DNA binding element for AP-1. The activation and plasm of bronchial epithelial cells shows positive immunoreactivity
binding of AP-1 to the eGPx promoter is a possible mechanism for to nitrotyrosine (red; arrow); g, goblet cell; bm, basement membrane.
eGPx mRNA and protein induction. I␬B, inhibitor of NF-␬B. Magnification, ⫻40; hematoxylin counterstaining.

AJP-Lung Cell Mol Physiol • VOL 283 • AUGUST 2002 • www.ajplung.org

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L252 INVITED REVIEW

gene products can form homodimeric (Jun-Jun) or het- thione peroxidase. Am J Physiol Cell Physiol 266: C367–C375,
erodimeric (Jun-Fos) complexes (100). The 5⬘-flanking 1994.
4. Babior BM. Oxygen-dependent microbial killing by phago-
region of the eGPx gene contains the DNA-binding cytes. N Engl J Med 298: 659–668, 1978.
element for AP-1 (137) (Fig. 3). Cigarette smoke in- 5. Barnes PJ and Belvisi MG. Nitric oxide and lung disease.
creases AP-1 DNA binding in human epithelial cells in Thorax 48: 1034–1043, 1993.
vivo (99). Based on our results and those of others, 6. Bast A, Haenen GR, and Doelman CJ. Oxidants and anti-
oxidants: state of the art. Am J Med 91: 2S–13S, 1991.
we propose that increased formation of ROS and 7. Becker K, Gui M, and Schirmer RH. Inhibition of human
RNS in inflammatory cells and epithelium leads to glutathione reductase by S-nitrosoglutathione. Eur J Biochem
alterations in the redox system, sensed by the epi- 234: 472–478, 1995.
thelial cells, and leads to the activation of transcrip- 8. Beckman JS and Koppenol WH. Nitric oxide, superoxide,
tion factors such as STATs and AP-1 and down- and peroxynitrite: the good, the bad, and the ugly. Am J Physiol
Cell Physiol 271: C1424–C1437, 1996.
stream target gene expression. 9. Beutler E and Matsumoto F. Ethnic variation in red cell
Function. On the basis of our studies and the studies glutathione peroxidase activity. Blood 46: 103–110, 1975.
of others, we have generated a model for the function of 10. Bonni A, Frank DA, Schindler C, and Greenberg ME.
eGPx in lung diseases associated with oxidative stress Characterization of a pathway for ciliary neurotrophic factor
(Fig. 3). NO is produced in mammalian airways, and signaling to the nucleus. Science 262: 1575–1579, 1993.
11. Brown DM, Beswick PH, Bell KS, and Donaldson K. De-
increased levels are found in many inflammatory lung pletion of glutathione and ascorbate in lung lining fluid by
diseases such as asthma and hyperoxia (28, 30, 61). respirable fibres. Ann Occup Hyg 44: 101–108, 2000.
Inflammation leads to increased levels of ROS (Fig. 4, 12. Buhl R, Jaffe HA, Holroyd KJ, Wells FB, Mastrangeli A,
pathway a). NO reacts slowly with O2 to form the Saltini C, Cantin AM, and Crystal RG. Systemic glutathione
deficiency in symptom-free HIV-seropositive individuals. Lan-
cytotoxic compound NO2⫺ or very rapidly with O2⫺䡠 to cet 2: 1294–1298, 1989.
form ONOO⫺ (Fig. 4, pathway b) (51). This results in 13. Bunnell E and Pacht ER. Oxidized glutathione is increased
tyrosine nitration in lung tissue (Fig. 5 and Fig. 4, in the alveolar fluid of patients with adult respiratory distress
pathway c). Thus when O2⫺䡠 is produced at high rates syndrome. Am Rev Respir Dis 148: 1174–1178, 1993.
during lung inflammation, NO may accelerate the for- 14. Busse R and Mulsch A. Induction of nitric oxide synthase by
cytokines in vascular smooth muscle cells. FEBS Lett 275:
mation of ONOO⫺, leading to tyrosine nitration and 87–90, 1990.
aggravate lung damage (61). The NO/O2⫺䡠 pathway in 15. Calhoun WJ, Reed HE, Moest DR, and Stevens CA. En-
vitro appears to be significantly shifted toward the hanced superoxide production by alveolar macrophages and
formation of GSNO in the presence of physiological air-space cells, airway inflammation, and alveolar macrophage
density changes after segmental antigen bronchoprovocation in
levels of GSH (Fig. 4, pathway d) (30, 45, 77). Recent allergic subjects. Am Rev Respir Dis 145: 317–325, 1992.
studies have shown that GPx (Fig. 4, pathway e) can 16. Candeias LP, Patel KB, Stratford MR, and Wardman P.
protect against NO-mediated protein oxidation (48) Free hydroxyl radicals are formed on reaction between the
and can reduce GSNO. Thus the increased eGPx in neutrophil-derived species superoxide anion and hypochlorous
lung inflammation may have two functions: reduction acid. FEBS Lett 333: 151–153, 1993.
17. Cantin AM, Hubbard RC, and Crystal RG. Glutathione
of ROS (e.g., H2O2 and O2⫺䡠) and possibly protection deficiency in the epithelial lining fluid of the lower respiratory
against NO-mediated protein oxidation by regulation tract in idiopathic pulmonary fibrosis. Am Rev Respir Dis 139:
of RSNO levels. 370–372, 1989.
In conclusion, lung diseases occur in response to 18. Cantin AM, North SL, Fells GA, Hubbard RC, and Crystal
RG. Oxidant-mediated epithelial cell injury in idiopathic pul-
oxidative and nitrosative stress. The lung’s ability to monary fibrosis. J Clin Invest 79: 1665–1673, 1987.
respond to oxidative stress depends largely on its ca- 19. Cantin AM, North SL, Hubbard RC, and Crystal RG.
pacity to upregulate protective antioxidants. The anti- Normal alveolar epithelial lining fluid contains high levels of
oxidant responses to lung disease vary widely, but the glutathione. J Appl Physiol 63: 152–157, 1987.
upregulation of eGPx in oxidant-mediated lung dis- 20. Cantin PA. Oxidant and antioxidants in lung injury. In: Lam
and Other Diseases Characterized by Smooth Muscle Prolifera-
eases is likely important to defend airway surfaces tion, edited by Moss J. New York: Dekker, 1999, p. 519–531.
from ROS and RNS injury. 21. Carp H, Miller F, Hoidal JR, and Janoff A. Potential mech-
anism of emphysema: alpha 1-proteinase inhibitor recovered
We thank J. H. J. M. van Krieken, F. B. J. M. Thunnissen, P. N. R. from lungs of cigarette smokers contains oxidized methionine
Dekhuijzen, D. Roos, A. Bast, and P. Scheepers for helpful discussion and has decreased elastase inhibitory capacity. Proc Natl Acad
and comments. Sci USA 79: 2041–2045, 1982.
This work was supported in part by National Heart, Lung, and 22. Chance B, Sies H, and Boveris A. Hydroperoxide metabo-
Blood Institute Grant HL-60917. lism in mammalian organs. Physiol Rev 59: 527–605, 1979.
23. Chu FF, Doroshow JH, and Esworthy RS. Expression,
REFERENCES characterization, and tissue distribution of a new cellular sele-
nium-dependent glutathione peroxidase, GSHPx-GI. J Biol
1. Arteel GE, Briviba K, and Sies H. Protection against per- Chem 268: 2571–2576, 1993.
oxynitrite. FEBS Lett 445: 226–230, 1999. 24. Clark AG and Debnam P. Inhibition of glutathione S-trans-
2. Avissar N, Finkelstein JN, Horowitz S, Willey JC, Coy E, ferases from rat liver by S-nitroso-L-glutathione. Biochem
Frampton MW, Watkins RH, Khullar P, Xu YL, and Cohen Pharmacol 37: 3199–3201, 1988.
HJ. Extracellular glutathione peroxidase in human lung epi- 25. Clark JM and Lambertsen CJ. Pulmonary oxygen toxicity: a
thelial lining fluid and in lung cells. Am J Physiol Lung Cell Mol review. Pharmacol Rev 23: 37–133, 1971.
Physiol 270: L173–L182, 1996. 26. Cochrane CG, Spragg R, and Revak SD. Pathogenesis of
3. Avissar N, Ornt DB, Yagil Y, Horowitz S, Watkins RH, the adult respiratory distress syndrome. Evidence of oxidant
Kerl EA, Takahashi K, Palmer IS, and Cohen HJ. Human activity in bronchoalveolar lavage fluid. J Clin Invest 71: 754–
kidney proximal tubules are the main source of plasma gluta- 761, 1983.

AJP-Lung Cell Mol Physiol • VOL 283 • AUGUST 2002 • www.ajplung.org

Downloaded from journals.physiology.org/journal/ajplung (179.202.020.007) on March 17, 2020.

 INVITED REVIEW L253

27. Comhair SA, Bhathena PR, Dweik RA, Kavuru M, and 48. Fu Y, McCormick CC, Roneker C, and Lei XG. Lipopoly-
Erzurum SC. Rapid loss of superoxide dismutase activity saccharide and interferon-␥-induced nitric oxide production and
during antigen-induced asthmatic response. Lancet 355: 624, protein oxidation in mouse peritoneal macrophages are affected
2000. by glutathione peroxidase-1 gene knockout. Free Radic Biol
28. Comhair SA, Bhathena PR, Farver C, Thunnissen FB, Med 31: 450–459, 2001.
and Erzurum SC. Extracellular glutathione peroxidase induc- 49. Gardner HW. Oxygen radical chemistry of polyunsaturated
tion in asthmatic lungs: evidence for redox regulation of expres- fatty acids. Free Radic Biol Med 7: 65–86, 1989.
sion in human airway epithelial cells. FASEB J 15: 70–78, 50. Gardner PR and Fridovich I. Superoxide sensitivity of the
2001. Escherichia coli aconitase. J Biol Chem 266: 19328–19333,
29. Comhair SA, Lewis MJ, Bhathena PR, Hammel JP, and 1991.
Erzurum SC. Increased glutathione and glutathione peroxi- 51. Gaston B, Drazen JM, Loscalzo J, and Stamler JS. The
dase in lungs of individuals with chronic beryllium disease. biology of nitrogen oxides in the airways. Am J Respir Crit Care
Am J Respir Crit Care Med 159: 1824–1829, 1999. Med 149: 538–551, 1994.
30. Comhair SA, Thomassen MJ, and Erzurum SC. Differen- 52. Goldstein S, Czapski G, Lind J, and Merenyi G. Tyrosine
tial induction of extracellular glutathione peroxidase and nitric nitration by simultaneous generation of 䡠 NO and O2 under
oxide synthase 2 in airways of healthy individuals exposed to physiological conditions. How the radicals do the job. J Biol
100% O2 or cigarette smoke. Am J Respir Cell Mol Biol 23: Chem 275: 3031–3036, 2000.
350–354, 2000. 53. Halliwell B. Free radicals, antioxidants, and human disease:
31. Conner EM and Grisham MB. Inflammation, free radicals, curiosity, cause, or consequence? Lancet 344: 721–724, 1994.
and antioxidants. Nutrition 12: 274–277, 1996. 54. Halliwell B and Gutteridge JM. Role of free radicals and
32. Crapo JD, Barry BE, Foscue HA, and Shelburne J. Struc- catalytic metal ions in human disease: an overview. Methods
tural and biochemical changes in rat lungs occurring during Enzymol 186: 1–85, 1990.
exposures to lethal and adaptive doses of oxygen. Am Rev 55. Han J, Stamler JS, Li H, and Griffith OW. Inhibition of
Respir Dis 122: 123–143, 1980. ␥-glutamylcysteine synthetase by S-nitrosylation. In: The Biol-
33. Crystal RG. Oxidants and respiratory tract epithelial injury: ogy of Nitric Oxide Part 5, edited by Stamler JS. Colchester,
pathogenesis and strategies for therapeutic intervention. Am J England: Portland Press, 1996, p. 115.
Med 91: 39S–44S, 1991. 56. Harrison DG, Chapman MP, Christy JP, and Marcus ML.
34. Davies KJ. Oxidative stress: the paradox of aerobic life. Bio- Studies of functional site of origin of native coronary collaterals.
chem Soc Symp 61: 1–31, 1995. Am J Physiol Heart Circ Physiol 251: H1217–H1224, 1986.
35. De Raeve HR, Thunnissen FB, Kaneko FT, Guo FH, 57. Hazen SL, Zhang R, Shen Z, Wu W, Podrez EA, MacPher-
Lewis M, Kavuru MS, Secic M, Thomassen MJ, and Er- son JC, Schmitt D, Mitra SN, Mukhopadhyay C, Chen Y,
zurum SC. Decreased Cu,Zn-SOD activity in asthmatic airway Cohen PA, Hoff HF, and Abu-Soud HM. Formation of nitric
oxide-derived oxidants by myeloperoxidase in monocytes: path-
epithelium: correction by inhaled corticosteroid in vivo. Am J
ways for monocyte-mediated protein nitration and lipid peroxi-
Physiol Lung Cell Mol Physiol 272: L148–L154, 1997.
dation in vivo. Circ Res 85: 950–958, 1999.
36. Deisseroth A and Dounce AL. Catalase: physical and chem-
58. Heffner JE and Repine JE. Pulmonary strategies of antiox-
ical properties, mechanism of catalysis, and physiological role.
idant defense. Am Rev Respir Dis 140: 531–554, 1989.
Physiol Rev 50: 319–375, 1970.
59. Henry Y, Ducrocq C, Drapier JC, Servent D, Pellat C, and
37. Deneke SM and Fanburg BL. Regulation of cellular gluta-
Guissani A. Nitric oxide, a biological effector. Electron para-
thione. Am J Physiol Lung Cell Mol Physiol 257: L163–L173,
magnetic resonance detection of nitrosyl-iron-protein com-
1989.
plexes in whole cells. Eur Biophys J 20: 1–15, 1991.
38. Doelman CJ and Bast A. Oxygen radicals in lung pathology.
60. Hogg N, Singh RJ, and Kalyanaraman B. The role of glu-
Free Radic Biol Med 9: 381–400, 1990. tathione in the transport and catabolism of nitric oxide. FEBS
39. Duan YJ, Komura S, Fiszer-Szafarz B, Szafarz D, and Lett 382: 223–228, 1996.
Yagi K. Purification and characterization of a novel monomeric 61. Honda K, Kobayashi H, Hataishi R, Hirano S, Fukuyama
glutathione peroxidase from rat liver. J Biol Chem 263: 19003– N, Nakazawa H, and Tomita T. Inhaled nitric oxide reduces
19008, 1988. tyrosine nitration after lipopolysaccharide instillation into
40. Eiserich JP, Cross CE, Jones AD, Halliwell B, and van lungs of rats. Am J Respir Crit Care Med 160: 678–688, 1999.
der Vliet A. Formation of nitrating and chlorinating species by 62. Janssen YM, Marsh JP, Absher MP, Hemenway D, Vacek
reaction of nitrite with hypochlorous acid. A novel mechanism PM, Leslie KO, Borm PJ, and Mossman BT. Expression of
for nitric oxide-mediated protein modification. J Biol Chem 271: antioxidant enzymes in rat lungs after inhalation of asbestos or
19199–19208, 1996. silica. J Biol Chem 267: 10625–10630, 1992.
41. Eiserich JP, Hristova M, Cross CE, Jones AD, Freeman 63. Johnson D and Travis J. The oxidative inactivation of human
BA, Halliwell B, and van der Vliet A. Formation of nitric alpha-1-proteinase inhibitor. Further evidence for methionine
oxide-derived inflammatory oxidants by myeloperoxidase in at the reactive center. J Biol Chem 254: 4022–4026, 1979.
neutrophils. Nature 391: 393–397, 1998. 64. Kanazawa H, Kurihara N, Hirata K, and Takeda T. The
42. Erzurum SC, Danel C, Gillissen A, Chu CS, Trapnell BC, role of free radicals in airway obstruction in asthmatic patients.
and Crystal RG. In vivo antioxidant gene expression in hu- Chest 100: 1319–1322, 1991.
man airway epithelium of normal individuals exposed to 100% 65. Kelm M. Nitric oxide metabolism and breakdown. Biochim
O2. J Appl Physiol 75: 1256–1262, 1993. Biophys Acta 1411: 273–289, 1999.
43. Folz RJ, Guan J, Seldin MF, Oury TD, Enghild JJ, and 66. Kharitonov VG, Sundquist AR, and Sharma VS. Kinetics
Crapo JD. Mouse extracellular superoxide dismutase: primary of nitric oxide autoxidation in aqueous solution. J Biol Chem
structure, tissue-specific gene expression, chromosomal local- 269: 5881–5883, 1994.
ization, and lung in situ hybridization. Am J Respir Cell Mol 67. Kim KK, Whitin JC, Sukhova NM, and Cohen HJ. Increase
Biol 17: 393–403, 1997. in extracellular glutatione peroxidase in plasma and lungs of
44. Foote CS, Goyne TE, and Lehrer RI. Assessment of chlori- mice exposed to hyperoxia. Pediatr Res 46: 715–721, 1999.
nation by human neutrophils. Nature 301: 715–716, 1983. 68. Kingsley PD, Whitin JC, Cohen HJ, and Palis J. Develop-
45. Freedman JE, Frei B, Welch GN, and Loscalzo J. Gluta- mental expression of extracellular glutathione peroxidase sug-
thione peroxidase potentiates the inhibition of platelet function gests antioxidant roles in deciduum, visceral yolk sac, and skin.
by S-nitrosothiols. J Clin Invest 96: 394–400, 1995. Mol Reprod Dev 49: 343–355, 1998.
46. Freeman BA and Crapo JD. Biology of disease: free radicals 69. Klatt P, Molina EP, De Lacoba MG, Padilla CA, Martinez-
and tissue injury. Lab Invest 47: 412–426, 1982. Galesteo E, Barcena JA, and Lamas S. Redox regulation of
47. Fridovich I. The biology of oxygen radicals. Science 201: 875– c-Jun DNA binding by reversible S-glutathiolation. FASEB J
880, 1978. 13: 1481–1490, 1999.

AJP-Lung Cell Mol Physiol • VOL 283 • AUGUST 2002 • www.ajplung.org

Downloaded from journals.physiology.org/journal/ajplung (179.202.020.007) on March 17, 2020.

L254 INVITED REVIEW

70. Klebanoff SJ and Hamon CB. Role of myeloperoxidase-me- 93. Peterson JD, Herzenberg LA, Vasquez K, and Walten-
diated antimicrobial systems in intact leukocytes. J Reticuloen- baugh C. Glutathione levels in antigen-presenting cells mod-
dothel Soc 12: 170–196, 1972. ulate Th1 versus Th2 response patterns. Proc Natl Acad Sci
71. Lakari E, Paakko P, and Kinnula VL. Manganese superox- USA 95: 3071–3076, 1998.
ide dismutase, but not CuZn superoxide dismutase, is highly 94. Pietarinen-Runtti P, Raivio KO, Saksela M, Asikainen
expressed in the granulomas of pulmonary sarcoidosis and TM, and Kinnula VL. Antioxidant enzyme regulation and
extrinsic allergic alveolitis. Am J Respir Crit Care Med 158: resistance to oxidants of human bronchial epithelial cells cul-
589–596, 1998. tured under hyperoxic conditions. Am J Respir Cell Mol Biol 19:
72. Liu RH, Baldwin B, Tennant BC, and Hotchkiss JH. Ele- 286–292, 1998.
vated formation of nitrate and N-nitrosodimethylamine in 95. Podrez EA, Schmitt D, Hoff HF, and Hazen SL. Myeloper-
woodchucks (Marmota monax) associated with chronic wood- oxidase-generated reactive nitrogen species convert LDL into
chuck hepatitis virus infection. Cancer Res 51: 3925–3929, an atherogenic form in vitro. J Clin Invest 103: 1547–1560,
1991. 1999.
73. Lowenstein CJ and Snyder SH. Nitric oxide, a novel biologic 96. Powis G. Metabolism and reactions of quinoid anticancer
messenger. Cell 70: 705–707, 1992. agents. Pharmacol Ther 35: 57–162, 1987.
74. MacNee W and Rahman I. Oxidants/antioxidants in idio- 97. Powis G. Free radical formation by antitumor quinones. Free
pathic pulmonary fibrosis. Thorax 50, Suppl 1: S53–S58, 1995.
Radic Biol Med 6: 63–101, 1989.
75. Marletta MA. Mammalian synthesis of nitrite, nitrate, nitric
98. Rahman I and MacNee W. Role of transcription factors in
oxide, and N-nitrosating agents. Chem Res Toxicol 1: 249–257,
inflammatory lung diseases. Thorax 53: 601–612, 1998.
1988.
99. Rahman I and MacNee W. Lung glutathione and oxidative
76. Marletta MA. Nitric oxide synthase structure and mechanism.
stress: implications in cigarette smoke-induced airway disease.
J Biol Chem 268: 12231–12234, 1993.
77. Mayer B, Schrammel A, Klatt P, Koesling D, and Schmidt Am J Physiol Lung Cell Mol Physiol 277: L1067–L1088, 1999.
K. Peroxynitrite-induced accumulation of cyclic GMP in endo- 100. Rahman I and MacNee W. Oxidative stress and regulation of
thelial cells and stimulation of purified soluble guanylyl cy- glutathione in lung inflammation. Eur Respir J 16: 534–554,
clase. Dependence on glutathione and possible role of S-nitro- 2000.
sation. J Biol Chem 270: 17355–17360, 1995. 101. Ramezanian MS, Padmaja S, and Koppenol WH. Nitration
78. McCay PB. Vitamin E: interactions with free radicals and and hydroxylation of phenolic compounds by peroxynitrite.
ascorbate. Annu Rev Nutr 5: 323–340, 1985. Chem Res Toxicol 9: 232–240, 1996.
79. McCord JM and Fridovich I. Superoxide dismutase. An 102. Reid TJ III, Murthy MR, Sicignano A, Tanaka N, Musick
enzymic function for erythrocuprein (hemocuprein). J Biol WD, and Rossmann MG. Structure and heme environment of
Chem 244: 6049–6055, 1969. beef liver catalase at 2.5 A resolution. Proc Natl Acad Sci USA
80. Meister A and Anderson ME. Glutathione. Annu Rev Bio- 78: 4767–4771, 1981.
chem 52: 711–760, 1983. 103. Ren B, Huang W, Akesson B, and Ladenstein R. The
81. Melloni B, Lefebvre MA, Bonnaud F, Vergnenegre A, crystal structure of seleno-glutathione peroxidase from human
Grossin L, Rigaud M, and Cantin A. Antioxidant activity in plasma at 2.9 A resolution. J Mol Biol 268: 869–885, 1997.
bronchoalveolar lavage fluid from patients with lung cancer. 104. Repine JE, Bast A, and Lankhorst I. Oxidative stress in
Am J Respir Crit Care Med 154: 1706–1711, 1996. chronic obstructive pulmonary disease. Oxidative stress study
82. Meyer KC. Beryllium and lung disease. Chest 106: 942–946, group. Am J Respir Crit Care Med 156: 341–357, 1997.
1994. 105. Ross D, Norbeck K, and Moldeus P. The generation and
83. Meyer M, Schreck R, and Baeuerle PA. H2O2 and antioxi- subsequent fate of glutathionyl radicals in biological systems.
dants have opposite effects on activation of NF-␬B and AP-1 in J Biol Chem 260: 15028–15032, 1985.
intact cells: AP-1 as secondary antioxidant-responsive factor. 106. Rowley DA and Halliwell B. Formation of hydroxyl radicals
EMBO J 12: 2005–2015, 1993. from hydrogen peroxide and iron salts by superoxide- and
84. Misso NL, Peacock CD, Watkins DN, and Thompson PJ. ascorbate-dependent mechanisms: relevance to the pathology of
Nitrite generation and antioxidant effects during neutrophil rheumatoid disease. Clin Sci (Colch) 64: 649–653, 1983.
apoptosis. Free Radic Biol Med 28: 934–943, 2000. 107. Schraufstatter UI. Oxidants: types, sources and mechanisms
85. Moro MA, Darley-Usmar VM, Goodwin DA, Read NG, of injury. In: The Lung: Scientific Foundations, edited by Crys-
Zamora-Pino R, Feelisch M, Radomski MW, and Moncada tal JW, Weible ER, and Barnes PJ. New York: Raven, 1996.
S. Paradoxical fate and biological action of peroxynitrite on 108. Schreck R and Baeuerle PA. NF-␬B as inducible transcrip-
human platelets. Proc Natl Acad Sci USA 91: 6702–6706, 1994. tional activator of the granulocyte-macrophage colony-stimu-
86. Morrison D, Rahman I, Lannan S, and MacNee W. Epithe- lating factor gene. Mol Cell Biol 10: 1281–1286, 1990.
lial permeability, inflammation, and oxidant stress in the air 109. Schwartz SE. Properties, transformations, and fates. In:
spaces of smokers. Am J Respir Crit Care Med 159: 473–479,
Trace Atmospheric Constituents, edited by Schwartz SE. New
1999.
York: Wiley, 1983.
87. Mossman BT. Signal transduction by oxidants: look who’s
110. Sen CK and Packer L. Antioxidant and redox regulation of
talking. Free Radic Biol Med 28: 1315–1316, 2000.
gene transcription. FASEB J 10: 709–720, 1996.
88. Mossman BT and Churg A. Mechanisms in the pathogenesis
of asbestosis and silicosis. Am J Respir Crit Care Med 157: 111. Shull S, Heintz NH, Periasamy M, Manohar M, Janssen
1666–1680, 1998. YM, Marsh JP, and Mossman BT. Differential regulation of
89. Nathan C and Xie QW. Regulation of biosynthesis of nitric antioxidant enzymes in response to oxidants. J Biol Chem 266:
oxide. J Biol Chem 269: 13725–13728, 1994. 24398–24403, 1991.
90. Palmer RM, Rees DD, Ashton DS, and Moncada S. 112. Simon AR, Rai U, Fanburg BL, and Cochran BH. Activa-
L-arginine is the physiological precursor for the formation of tion of the JAK-STAT pathway by reactive oxygen species.
nitric oxide in endothelium-dependent relaxation. Biochem Bio- Am J Physiol Cell Physiol 275: C1640–C1652, 1998.
phys Res Commun 153: 1251–1256, 1988. 113. Singh S and Evans TW. Nitric oxide, the biological mediator
91. Parks NJ, Krohn KJ, Mathis CA, Chasko JH, Geiger KR, of the decade: fact or fiction? Eur Respir J 10: 699–707, 1997.
Gregor ME, and Peek NF. Nitrogen-13-labeled nitrite and 114. Smith LJ, Houston M, and Anderson J. Increased levels of
nitrate: distribution and metabolism after intratracheal admin- glutathione in bronchoalveolar lavage fluid from patients with
istration. Science 212: 58–60, 1981. asthma. Am Rev Respir Dis 147: 1461–1464, 1993.
92. Patel RP, McAndrew J, Sellak H, White CR, Jo H, Free- 115. Smith LJ, Shamsuddin M, Sporn PH, Denenberg M, and
man BA, and Darley-Usmar VM. Biological aspects of reac- Anderson J. Reduced superoxide dismutase in lung cells of
tive nitrogen species. Biochim Biophys Acta 1411: 385–400, patients with asthma. Free Radic Biol Med 22: 1301–1307,
1999. 1997.

AJP-Lung Cell Mol Physiol • VOL 283 • AUGUST 2002 • www.ajplung.org

Downloaded from journals.physiology.org/journal/ajplung (179.202.020.007) on March 17, 2020.

 INVITED REVIEW L255

116. Staal FJ, Roederer M, and Herzenberg LA. Intracellular 128. Wagner BJ, Hayes TE, Hoban CJ, and Cochran BH. The
thiols regulate activation of nuclear factor kappa B and tran- SIF binding element confers sis/PDGF inducibility onto the
scription of human immunodeficiency virus. Proc Natl Acad Sci c-fos promoter. EMBO J 9: 4477–4484, 1990.
USA 87: 9943–9947, 1990. 129. Webster NR and Nunn JF. Molecular structure of free radi-
117. Stamler JS, Singel DJ, and Loscalzo J. Biochemistry of cals and their importance in biological reactions. Br J Anaesth
nitric oxide and its redox-activated forms. Science 258: 1898– 60: 98–108, 1988.
1902, 1992. 130. Weiss SJ, Test ST, Eckmann CM, Roos D, and Regiani S.
118. Sun Y and Oberley LW. Redox regulation of transcriptional Brominating oxidants generated by human eosinophils. Science
activators. Free Radic Biol Med 21: 335–348, 1996. 234: 200–203, 1986.
119. Tanswell AK and Freeman BA. Antioxidant therapy in crit- 131. Wink DA, Hanbauer I, Grisham MB, Laval F, Nims RW,
ical care medicine. New Horiz 3: 330–341, 1995. Laval J, Cook J, Pacelli R, Liebmann J, Krishna M, Ford
120. Tham DM, Whitin JC, Kim KK, Zhu SX, and Cohen HJ. PC, and Mitchell JB. Chemical biology of nitric oxide: regu-
Expression of extracellular glutathione peroxidase in human lation and protective and toxic mechanisms. Curr Top Cell
and mouse gastrointestinal tract. Am J Physiol Gastrointest Regul 34: 159–187, 1996.
Liver Physiol 275: G1463–G1471, 1998. 132. Wink DA, Nims RW, Darbyshire JF, Christodoulou D,
121. Thomas EL, Learn DB, Jefferson MM, and Weatherred Hanbauer I, Cox GW, Laval F, Laval J, Cook JA, Krishna
W. Superoxide-dependent oxidation of extracellular reducing MC, Degraff WG, and Mitchell JB. Reaction kinetics for
agents by isolated neutrophils. J Biol Chem 263: 2178–2186, nitrosation of cysteine and glutathione in aerobic nitric oxide
1988. solutions at neutral pH. Insights into the fate and physiological
effects of intermediates generated in the NO/O2 reaction. Chem
122. Vallyathan V and Shi X. The role of oxygen free radicals in
Res Toxicol 7: 519–525, 1994.
occupational and environmental lung diseases. Environ Health
133. Winterbourn CC and Munday R. Glutathione-mediated re-
Perspect 105, Suppl 1: 165–177, 1997.
dox cycling of alloxan. Mechanisms of superoxide dismutase
123. Van Acker SA, Koymans LM, and Bast A. Molecular phar-
inhibition and of metal-catalyzed OH formation. Biochem Phar-
macology of vitamin E: structural aspects of antioxidant activ-
macol 38: 271–277, 1989.
ity. Free Radic Biol Med 15: 311–328, 1993. 134. Wright DT, Cohn LA, Li H, Fischer B, Li CM, and Adler
124. Van der Vliet A, Eiserich JP, Halliwell B, and Cross CE. KB. Interactions of oxygen radicals with airway epithelium.
Formation of reactive nitrogen species during peroxidase-cata- Environ Health Perspect 102, Suppl 10: 85–90, 1994.
lyzed oxidation of nitrite. A potential additional mechanism of 135. Wu W, Chen Y, and Hazen SL. Eosinophil peroxidase ni-
nitric oxide-dependent toxicity. J Biol Chem 272: 7617–7625, trates protein tyrosyl residues. Implications for oxidative dam-
1997. age by nitrating intermediates in eosinophilic inflammatory
125. Van der Vliet A, Eiserich JP, Shigenaga MK, and Cross disorders. J Biol Chem 274: 25933–25944, 1999.
CE. Reactive nitrogen species and tyrosine nitration in the 136. Wu W, Samoszuk MK, Comhair SA, Thomassen MJ,
respiratory tract: epiphenomena or a pathobiologic mechanism Farver CF, Dweik RA, Kavuru MS, Erzurum SC, and
of disease? Am J Respir Crit Care Medic 160: 1–9, 1999. Hazen SL. Eosinophils generate brominating oxidants in al-
126. Van der Vliet A, Hoen PA, Wong PS, Bast A, and Cross lergen-induced asthma. J Clin Invest 105: 1455–1463, 2000.
CE. Formation of S-nitrosothiols via direct nucleophilic nitro- 137. Yoshimura S, Suemizu H, Taniguchi Y, Arimori K, Ka-
sation of thiols by peroxynitrite with elimination of hydrogen wabe N, and Moriuchi T. The human plasma glutathione
peroxide. J Biol Chem 273: 30255–30262, 1998. peroxidase-encoding gene: organization, sequence and localiza-
127. Van der Vliet A, Smith D, O’Neill CA, Kaur H, Darley- tion to chromosome 5q32. Gene 145: 293–297, 1994.
Usmar V, Cross CE, and Halliwell B. Interactions of per- 138. Zhang Y, Marcillat O, Giulivi C, Ernster L, and Davies
oxynitrite with human plasma and its constituents: oxidative KJ. The oxidative inactivation of mitochondrial electron trans-
damage and antioxidant depletion. Biochem J 303: 295–301, port chain components and ATPase. J Biol Chem 265: 16330–
1994. 16336, 1990.

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