Am J Physiol Lung Cell Mol Physiol 314: L743–L756, 2018.

First published January 4, 2018; doi:10.1152/ajplung.00373.2017.

REVIEW

Effects of cigarette smoke on pulmonary endothelial cells

X Qing Lu,1,2 Eric Gottlieb,2 and Sharon Rounds1,2
1
Vascular Research Laboratory, Providence Veterans Affairs Medical Center, Providence, Rhode Island; and 2Department of
Medicine, Alpert Medical School of Brown University, Providence, Rhode Island
Submitted 18 August 2017; accepted in final form 22 December 2017

Lu Q, Gottlieb E, Rounds S. Effects of cigarette smoke on pulmonary

endothelial cells. Am J Physiol Lung Cell Mol Physiol 314: L743–L756, 2018. First
published January 4, 2018; doi:10.1152/ajplung.00373.2017.—Cigarette smoking
is the leading cause of preventable disease and death in the United States.
Cardiovascular comorbidities associated with both active and secondhand cigarette
smoking indicate the vascular toxicity of smoke exposure. Growing evidence
supports the injurious effect of cigarette smoke on pulmonary endothelial cells and
the roles of endothelial cell injury in development of acute respiratory distress
syndrome (ARDS), emphysema, and pulmonary hypertension. This review sum-
marizes results from studies of humans, preclinical animal models, and cultured
endothelial cells that document toxicities of cigarette smoke exposure on pulmo-
nary endothelial cell functions, including barrier dysfunction, endothelial activation
and inflammation, apoptosis, and vasoactive mediator production. The discussion is
focused on effects of cigarette smoke-induced endothelial injury in the develop-
ment of ARDS, emphysema, and vascular remodeling in chronic obstructive
pulmonary disease.
acute respiratory distress syndrome; cigarette smoke; emphysema; endothelial
cells; pulmonary hypertension

INTRODUCTION also associated with increased risk of coronary artery disease

Cigarette smoking is the leading cause of preventable dis-
ease and death in the US, causing ⬎480,000 deaths per year in
the US, according to the 2014 Surgeon General’s report (136a).
Worldwide, the World Health Organization estimates that
tobacco use kills ⬎7 million people each year, 6 million of
whom are smokers while 890,000 die from the effects of
secondhand smoke exposure [World Health Organization To-
bacco Fact Sheet (150a)]. Although the incidence of tobacco
use in the US has decreased, 20.1% of adults still continued to
use tobacco products in 2015 (106). Thus cigarette smoking
remains a significant cause of morbidity and mortality.
Cigarette smoking has systemic effects, in addition to alter-
ing lung function. Causes of smoking-related deaths include
cancers, cardiovascular and metabolic diseases, pulmonary
diseases, and conditions related to pregnancy and birth [2014
Surgeon General’s report (136a)]. Chronic lung diseases
caused by cigarette smoking include chronic obstructive pul-
monary disease (COPD), idiopathic pulmonary fibrosis, and
asthma. COPD in particular is associated with systemic comor-
bidities (“the comorbidome”; 35). Among the systemic comor-
bidities, cardiovascular diseases, including pulmonary hyper-
tension, are the most prevalent and carry a high risk of
mortality (32). Furthermore, secondhand smoke exposure is
Address for reprint requests and other correspondence: Q. Lu, Vascular
Research Laboratory, Providence VA Medical Center, 830 Chalkstone Ave.,
Providence, RI 02908 (e-mail: Qing_Lu@brown.edu).
http://www.ajplung.org
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and other cardiovascular diseases (13). These cardiovascular
comorbidities of both mainstream and secondhand smoke ex-
posure suggest that toxic cigarette smoke constituents are
absorbed from the airways into the blood and that the inhala-
tion of cigarette smoke causes systemic toxicity.
A growing body of evidence indicates that cigarette smoke
alters vascular function, in addition to airways and immune
function (127). Voelkel and colleagues were among the first to
demonstrate lung endothelial cell apoptosis in human emphy-
sema [66; Imai et al. (59)] and in animal models of cigarette
smoke exposure [Marwick et al. (88)] and suggested the
“vascular hypothesis” for pathogenesis of emphysema (140).
Endothelial progenitor cell dysfunction has been recognized in
acute exacerbation of COPD, suggesting that cigarette smoke
(CS)-induced inadequate repair of the endothelium may also
contribute to COPD mortality (80).
CS is complex and includes ~4,500 components. Among
these, CO, nicotine, oxidants, fine particulate matter, aryl
hydrocarbons, and aldehydes such as formaldehyde and acro-
lein have potential for causing endothelial cell injury. Acrolein
is of particular interest since increased levels have been found
in blood and urine of smokers (7). Intraperitoneal administra-
tion of acrolein causes emphysema in rats (71), and intratra-
cheal instillation increases lung vascular permeability in mice
(82). A recent review summarized the known effects on lung
function of nicotine administered by electronic cigarettes (30).
It is now recognized that the lung endothelium is not a
passive barrier to gas exchange, but that it is metabolically
L743
L744 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM

active and is a key mediator of inflammation. Among the
functions of lung endothelium, barrier function is critical in
preventing extravasation of water, protein, and cells from the
circulation, thus maintaining normal gas exchange. A conse-
quence of injury to the lung endothelium is loss of barrier
function and increased-permeability pulmonary edema, the
pathophysiologic hallmark of acute respiratory distress syn-
drome (ARDS). ARDS is an important cause of mortality that
does not have specific treatment, despite great improvements in
supportive care. In this review we consider growing epidemi-
ologic evidence that cigarette smokers are at increased risk for
ARDS and are more susceptible to loss of lung endothelial
barrier function from other events associated with development
of ARDS (Table 1). In addition, we summarize evidence for
cigarette smoke effects on alveolar loss in emphysema and on
vascular remodeling and reactivity in pulmonary hypertension
associated with COPD. We review studies using cell and
animal models of lung endothelial toxicity caused by cigarette
smoke exposure, resulting in abnormal barrier function, in-
creased permeability pulmonary edema, endothelial cell apo-
ptosis, endothelial activation and inflammation, and endothe-
lial-dependent vascular remodeling.
EFFECT OF CIGARETTE SMOKING ON ENDOTHELIAL
PERMEABILITY AND ACTIVATION IN ARDS
CS Is a Priming Factor for ARDS in Human Subjects
CS increases susceptibility to lung infections and develop-
ment of ARDS in human subjects. Respiratory tract infections
and complications are common causes of morbidity and mor-
tality among smokers (136a). Smokers are more susceptible to
infections by human immunodeficiency virus (HIV; 107, 113),
Mycobacterium tuberculosis (27), Streptococcus pneumoniae
(15), and influenza A (150). Current smokers with community-
acquired pneumonia often progress to severe sepsis (15). In
addition to increased risk of infections, recent work has impli-
cated primary and secondhand smoke exposure as a risk factor
for the development of ARDS. Iribarren et al. extracted ARDS
cases from a Kaiser Permanente database of hospitalizations
(60). They found that current heavy smokers (⬎20 cigarettes
per day) had 5.7 times the risk of developing ARDS, with a
dose-response association in a multivariate regression. They
estimated that 50% of the risk of ARDS is attributable to
cigarette smoking. In a study of esophagectomy patients, a
history of smoking was one of two preoperative factors asso-
ciated with the later development of ARDS (132). Wacharasint
et al. found that in a cohort of patients admitted to a surgical
intensive care unit in Thailand, those who were active smokers
had a higher incidence of ARDS than former smokers, who in
turn had a higher incidence than never smokers (141). This
study also demonstrated a dose-response association in which
pack years correlated with risk of ARDS. Ware et al. evaluated
the cohort from the Validating Acute Lung Injury Biomarkers
for Diagnosis (VALID) study (147). Although they were test-
ing for an association between ambient ozone levels and risk of
ARDS, they found that in this critically ill population, ozone
exposure was associated with ARDS only in patients who
smoked (147). Furthermore, when ARDS survivors were fol-
lowed as in the ARDS Network (ARDSNET) Long-Term
Outcomes Study (ALTOS), those who smoked had a poorer
health-related quality of life (22).
There are a few studies that have not shown smoking to be
a significant factor. Gajic et al. examined a variety of variables
in a prospective study to determine which were associated with
the development of mild and severe ARDS (42). While char-
acteristics including alcohol abuse, obesity, and male sex were
significant, smoking was not associated with higher rates of

Table 1. Human studies on effects of cigarette smoke on development of ARDS

Authors Year Cohort Smoking Status Association

Iribarren et al. 2000 HMO database, extracted ARDS admissions Patient report Dose-response effect between cigarette
(60) smoking and risk of ARDS
Tandon et al. 2001 Esophagectomy patients in the United Patient report Smoking was associated with
(132) Kingdom postesophagectomy ARDS
Calfee et al. (26) 2011 Emergency department visits for blunt trauma Cotinine levels measured at Cotinine level correlated with risk of ARDS
admission
Hsieh et al. (57) 2014 ARDSNET cohort (2 studies) 4-(Methylnitrosamino)-1-(3- Similar risk of ARDS despite being younger
pyridyl)-1-butanol levels and healthier
Ware et al. (146) 2014 Brain-dead donors for Beta-Agonists for Medical record and interviews Current smokers had higher lung weights
Oxygenation in Lung Donors (BOLD) with next of kin than ever smokers, and ever smokers had
study higher lung weights than lifelong
nonsmokers (measure of edema)
Ware et al. (147) 2016 Validating Acute Lung Injury Biomarkers for Medical record/patient report Ozone exposure significantly associated
Diagnosis (VALID) study cohort with ARDS risk only in current smokers
Wacharasint et 2015 Thai-Surgical Intensive Care Unit (Thai- Medical record/patient report Smokers with higher number of pack years
al. (141) (poster) SICU) study databases and heavier smokers had higher incidence
of ARDS and longer ICU length of stay
Moazed et al. 2016 Volunteer smokers and nonsmokers Self-report Greater increase in total protein in smokers
(94) (measure of permeability) after exposure
to LPS, multiple biomarkers
García-Lucio et 2016 Lobectomy/pneumonectomy patients for Patient report Elevated ANGPT-2 in smokers
al. (43) solitary pulmonary nodule
Gajic et al. (42) 2011 Admitted patients with risk factors for ARDS Patient report No association between smoking and ARDS
Ferro et al. (38) 2010 Admitted trauma patients Patient report No difference in adverse outcomes when
stratified by smoking status
ANGPT-2, angiopoietin-2; ARDS, acute respiratory distress syndrome; ARDSNET, ARDS Network; HMO, health maintenance organization; ICU, intensive
care unit.

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 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM
ARDS (42). In a retrospective study of trauma patients, Ferro
et al. showed no association between smoking and various
adverse outcomes including ARDS, as well as sepsis, septic
shock, and mortality (38). Smoking also did not impact the
number of days of mechanical ventilation or intensive care unit
length of stay (38).
Notably, all the aforementioned studies relied on self-report
and/or chart review to determine smoking status. To better
avoid bias inherent in self-report and to capture secondhand
smoke exposure, a few studies have used biomarkers such as
cotinine and NNAL [4-(methylnitrosamino)-1-(3-pyridyl)-1-
butanol] to assess smoking status. Calfee et al. measured levels
of plasma cotinine and tracked the subsequent incidence of
ARDS (26). They found that patients in the highest quartile of
cotinine concentrations were 3.25 times as likely as those in the
lowest quartile to develop ARDS (26). Hsieh et al. performed
a secondary analysis of the ARDSNET Albuterol for the
Treatment of Acute Lung Injury (ALTA) and Omega-3 Fatty
Acid/Antioxidant Supplementation for Treating People with
Acute Lung Injury or Acute Respiratory Distress Syndrome
(Omega) studies. Smoking status was determined on the basis
of urine NNAL concentrations at the time of randomization in
the primary study (57). Smokers tended to be younger and
healthier and have fewer comorbidities and lower Acute
Physiology and Chronic Health Evaluation (APACHE)
scores than patients with undetectable NNAL, but they
nonetheless had equally severe lung injury. In these patients,
smoking nullified the protective benefit of overall better
health when they developed ARDS. Smokers were also
overrepresented in this sample of patients with ARDS com-
pared with national smoking rates (57).
CS increases alveolar-capillary barrier permeability and
inflammation in humans. The poor outcomes of ARDS among
smokers are likely due to inflammation, alveolar epithelial and
endothelial injury, and increased alveolar-capillary permeabil-
ity, all of which predispose to development of pulmonary
edema. In a study of donor lungs harvested with the inte-
ntion of transplant, Ware et al. (146) showed that lungs from
current smokers weighed more than lungs of former smo-
kers and that these were heavier than the lungs of never
smokers, all presumably because of edema. They found the
same pattern for efficiency of oxygenation [arterial partial
pressure of O2 (PaO2)/inspired O2 fraction (FIO2)] before har-
vest. Additionally, they found that a more extensive smoking
history was associated with worse results in a laboratory test of
alveolar fluid clearance (146), a measure of alveolar epithelial
cell function. Relative to the more permeable pulmonary
capillary endothelial barrier (pore radius of intercellular
junctions is ~40 – 80 Å), the alveolar epithelial barrier is less
permeable because of tight junctions of an estimated pore
radius of 4 –10 Å (133). Pulmonary clearance of inhaled
99m
technetium-labeled diethylenetriamine pentaacetic acid
(99mTc-DTPA) has been used to noninvasively assess alve-
olar epithelial barrier permeability in humans (14). Using
this technique, Jones et al. have documented increased
alveolar epithelial barrier permeability in cigarette smokers
(63). Taken together, CS increases alveolar epithelial barrier
permeability.
Whether CS exposure increases pulmonary endothelial bar-
rier permeability in humans has been controversial. A pilot
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L745
study on four smokers suggests that smokers may have de-
creased pulmonary capillary barrier permeability to urea (145).
In contrast, pulmonary endothelial denudation has been docu-
mented in smokers with COPD (9). Altered endothelial func-
tion has also been suggested by biomarkers in human subjects.
García-Lucio et al. (43) obtained pneumonectomy specimens
from a sample of patients with solitary nodules, who either had
COPD, were smokers with normal lung function, or were
nonsmokers with normal lung function. In these patients,
expression of angiopoietin-2 (ANGPT-2), as measured by
immunohistochemistry, was associated with increased pulmo-
nary artery vessel wall remodeling. In another cohort, they
found that plasma levels of ANGPT-2 were higher in current
smokers than in former smokers and that COPD patients
overall had higher levels of ANGPT-2 than nonsmokers.
Again, they demonstrated a dose-response association with
pack years (43). A similar association was demonstrated by
Petta et al. in healthy smokers with asthma (105). Since
ANGPT-2 is secreted by vascular endothelial cells in Weibel-
Palade bodies, these findings support the notion that cigarette
smoking alters endothelial cell function in humans.
Moazed et al. (94) demonstrated experimentally that after
exposure to inhaled lipopolysaccharide (LPS), smokers had a
greater increase in plasma IL-8, a known mediator of inflam-
mation in ARDS, compared with nonsmokers. These smokers
had increased bronchoalveolar lavage (BAL) protein, suggest-
ing increased alveolar-capillary permeability. They also had
elevated plasma matrix metalloproteinase-8 (MMP-8), a poten-
tial cause of lung matrix breakdown. MMP-8 is secreted by
neutrophils, which were elevated in total numbers and percent-
ages in BAL fluid. LPS inhalation caused an exaggerated
decrease in BAL VEGF, which has been shown to be de-
creased in ARDS patients and to increase with recovery (1).
Smokers had increased levels of soluble VEGF receptor 1
(VEGFR1), which inhibits VEGF and may contribute to in-
creased permeability (94). A recent study of mechanically
ventilated trauma patients also showed differences in the lung
microbiota of smokers and nonsmokers, at admission and after
48 h, which in turn affected expression of inflammatory mark-
ers and was associated with differences in incidence and
severity of ARDS (101).
In summary, with some exceptions, these clinical studies
present significant evidence that cigarette smoking potentiates
ARDS and worsens outcomes. Cigarette smoke exposure in-
creases both epithelial and endothelial barrier permeability at
baseline and upregulates inflammatory pathways in humans.
The role of inflammation in ARDS may differ among cases;
however, evidence from several ARDSNET trials suggests that
there are at least two subphenotypes of ARDS with distinct
biochemical and inflammatory profiles (25). This is clinically
significant because these subphenotypes respond differently to
liberal vs. conservative fluid strategies and to high vs. low
positive end-expiratory pressure (25, 36, 122). In clinical
practice, subphenotype determination is limited by the bio-
chemical assays available at the bedside. Future studies should
incorporate smoking history as a factor in the characterization
of ARDS subphenotypes and determine whether it can be used
to identify the subphenotype and influence treatment strategies
(25, 36, 122).
L746 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM

Both Brief and Subacute CS Exposures Increase Alveolar- result in endothelial cell injury and increased permeability. By
Capillary Barrier Permeability in Animal Models assessing blood concentrations of intratracheally instilled io-
dine-125 bovine serum albumin (125I-BSA), which has dimen-
Loss of alveolar-capillary barrier function is a hallmark of sions of 140 ⫻ 40 ⫻ 40 Å and is impermeable to a healthy
ARDS. By assessing pulmonary clearance of fluorescein iso- capillary endothelial barrier, Li et al. reported that intratracheal
thiocyanate-dextran (FITC-D; molecular radius at 22.2 Å), instillation of cigarette smoke condensate increases pulmonary
Burns et al. reported that CS exposure increases alveolar capillary barrier permeability in rats (78). They also suggested
epithelial barrier permeability in guinea pigs (23). When the that CS-induced increase in alveolar-capillary barrier permea-
alveolar epithelial barrier is compromised, components of CS bility was due to oxidative stress but not a consequence of lung
may pass through the leaky epithelial barrier and potentially inflammation (78). We have demonstrated that lung microvas-
cular endothelial cells (LMVEC) isolated from mice exposed
to CS had a greater permeability and incomplete recovery after
A challenges by LPS or thrombin (Fig. 1, A and B; 20). Our
results suggest that CS exposure may cause epigenetic changes
1.2
LPS in endothelial cells, which make these cells more vulnerable to
a second injury. Both brief (hours) and subacute (4 wk) CS
exposures increased BAL protein levels in guinea pigs (74).
Normalized Resistance

1.0
We have shown that acute CS preexposure followed by saline
∗ ∗ ∗ ∗ ∗ (control) caused lung edema; this effect was augmented in
ξ ∗
ξ response to a second injury (LPS or Pseudomonas) in mice
0.8 ξ ξ ξ ξ ξ
ξ (Fig. 2; 20, 84). To our surprise, CS exposure alone without
saline did not increase BAL protein levels or lung wet-to-dry
RA, V
0.6 weight ratio (data not shown). We suspect that the increased
RA, LPS
CS, V
alveolar-capillary permeability by CS exposure alone is so
CS, LPS covert in mice that it is not detectable by BAL protein content
0.4
or wet-to-dry lung weight ratio. We speculate that more sen-
0 5 10 15 20 sitive methods may be necessary to assess the effect of CS
Time (hours) alone on alveolar-capillary permeability in mice. Nevertheless,
B there is strong evidence suggesting that CS exposure increases
lung alveolar-capillary barrier permeability both in humans and
1.2 in other animal models.
Thrombin
Both Brief and Subacute CS Exposures Increase
Normalized Resistance

1.0 Susceptibility to Acute Lung Injury in Animal Models

ξ Similarly to the human studies described above, CS preex-

0.8 ξ posure delayed bacterial clearance and promoted lung inflam-
ξ
ξ
mation induced by Pseudomonas aeruginosa infection in mice
∗ ξ ξ ξ ξ
(33). Two weeks of CS exposure also enhanced alveolar
RA, V
0.6
RA, thrombin
inflammation and apoptosis induced by poly(I:C), a viral
CS, V
pathogen-associated molecular pattern (PAMP) in mice (65).
CS, thrombin
0.4
0 5 10 15 20
Time (hours) Fig. 1. Cigarette smoke (CS) increased lung microvascular endothelial cell
C (LMVEC) permeability in vitro. A and B: 6-wk-old male C57BL/6 mice were
exposed to room air (RA) or CS for 6 h. After overnight rest, LMVEC were
isolated from cortical lung tissues. Isolated and characterized LMVEC at
CSE passage 1 were plated onto electric cell-substrate impedance sensing (ECIS)
1.2
arrays at equal numbers of cells (2.5 ⫻ 105 cells per well). After overnight
attachment, cells were treated with vehicle (V), LPS (1 ␮g/ml), or thrombin (2
U/ml) for 20 h, and monolayer permeability was assessed by measuring
Normalized Resistance

electric resistance across the monolayers. Four independent experiments with

0.9 ∗ duplicated ECIS wells for each condition at each time were conducted. C:
∗
∗ primary human LMVEC were treated with vehicle (10% PBS) or varying
∗ concentrations of cigarette smoke extract (CSE) for indicated times, and
0.6 ∗ ∗ ∗ ∗ ∗
monolayer permeability was assessed by ECIS. Three independent experi-
V (10% PBS) ments with duplicated ECIS wells for each condition at each time were
5% CSE conducted. Data are presented as the means ⫾ SE of the normalized electrical
0.3 resistance at the selected time points relative to their initial resistance. ANOVA
7.5% CSE and Tukey-Kramer post hoc test were used to determine statistically significant
10% CSE difference across means among groups. *P ⬍ 0.05 vs. RA⫹V; ␰P ⬍ 0.05 vs.
0 RA⫹LPS (A) or RA⫹thrombin (B). Arrows indicate the time for addition of
0 2 4 6 8
treatments. [Reprinted with permission of the American Journal of Respiratory
Time (hours) Cell and Molecular Biology (20).]

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 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM L747
Four weeks of CS exposure exacerbated lung inflammation and chest trauma-induced lung inflammation and lung cell apopto-
compromised adaptive immunity to infection by Hemophilus sis in mice (142).
influenzae in mice (86). Two weeks of CS exposure did not Using two strains of inbred mice with different susceptibility
change basal BAL protein levels or neutrophil count but to CS-induced emphysema (48), we found that as little as 3– 6
significantly exacerbated mechanical ventilation-induced in- h of exposure to CS potentiated LPS-induced lung edema in
crease in BAL protein content and neutrophil influx in rats both C57BL/6 (Fig. 2A) and AKR mice (116). Similar exac-
(54). Similarly, CS exposure for 3– 4 wk aggravated blunt erbating effects were observed after 3 wk of preexposure to CS
(Fig. 3A). We also noted a strain difference in susceptibility to
CS priming for acute lung injury, with a greater effect in AKR
A C57BL/6 € mice (Fig. 3A; 116).
Taken together, studies of both human and preclinical ani-
1.2
mal models indicate that increased susceptibility to and sever-
∗ ity of acute lung injury/ARDS are important adverse health
1.0
ε consequences of CS exposure that need to be taken into
consideration when treating critically ill individuals. In addi-
BAL Protein

0.8
tion, there is evidence from humans and from animal studies
(mg/ml)

0.6
that CS exposure increases lung barrier permeability, through
effects on both epithelial and endothelial cells. In this review
0.4 we focus on mechanisms of CS effects on lung endothelial cell
permeability.
0.2
Cigarette Smoke Extract Increases Endothelial Monolayer
0 Permeability in Vitro
RA CS RA CS 6 hrs
Although the nature and concentrations of smoke compo-
ctrl LPS nents may differ between CS inhalation in vivo and aqueous
cigarette smoke extract (CSE) exposure of culture cells in
B C57BL/6
vitro, CSE exposure is a widely used and acceptable in vitro
∗ system to address effects of CS on specific cells and underlying
4.8 ε mechanisms (92, 155). As has been shown in human and
animal studies, exposure to CSE in vitro increases pulmonary
macrovascular and microvascular endothelial monolayer per-
4.4
meability (Fig. 1C; 20, 55). The underlying mechanism in-
Lung Wet/Dry

volves inhibition of RhoA and focal adhesion kinase (FAK)

4.0 signaling (84), microtubule depolymerization (16), histone
deacetylase-6 (HDAC6) activation (20), p38 activation (81),
3.6 ceramide upregulation (120), and disruption of intercellular
adhesion molecules, as depicted in Fig. 4.
3.2 Endothelial barrier integrity is regulated by adherens junc-
tions (AJs), F-actin cytoskeleton, microtubules (102), and focal
0
adhesion complexes (FAC; 138). It has been well documented
RA CS RA CS 6 hrs that RhoA is required for coordinated assembly of F-actin
fibers and that AJs and FAC are critical for establishment of
ctrl LPS
cell-cell contacts (21). RhoA activation also mediates in-
creased endothelial permeability induced by various edema-
C C57BL/6
genic agents, such as thrombin and histamine, by increasing
contractile F-actin stress fiber formation (136). The role of
2.5
RhoA signaling in CSE-induced endothelial monolayer perme-
€
ability has been controversial. Inhibition of Rho kinase has
2.0 been reported to attenuate CSE-induced increases in lung
BAL Protein

1.5
(mg/ml)

∗ Fig. 2. Effects of a brief cigarette smoke (CS) exposure on LPS- and

1.0 Pseudomonas-induced lung edema. Male 6-wk-old C57BL/6 were exposed to
room air (RA) or CS for 6 h. One hour after CS exposure, mice were intratrache-
ally administered with 2.5 mg/kg of LPS or 5 ⫻ 103 colony-forming units of P.
ε aeruginosa (strain PA103) or equal volume of saline as a control (ctrl). After 18
0.5
h, bronchoalveolar lavage (BAL) protein content (A and C) and lung wet-to-dry
weight ratio (B) were assessed. Four to six mice per group were used. εP ⬍ 0.05
0 CS/ctrl vs. RA/ctrl; *P ⬍ 0.05 RA/LPS vs. RA/ctrl; €P ⬍ 0.05 CS/LPS vs.
RA CS RA CS 6 hrs RA/LPS or RA/PA103. [Reprinted with permission of the American Journal of
Physiology Lung Cellular and Molecular Physiology (84) and the American
ctrl PA 103 Journal of Respiratory Cell and Molecular Biology (20).]

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L748 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM

A dize and activate RhoA through direct oxidation of two cys-

2.0 teine residues located in the redox-sensitive motif (2). Con-
€
versely, high levels of oxidants inhibit RhoA through forma-
tion of an intramolecular disulfide bridge that prevents GTP
BAL Protein

1.5
(mg/ml)

binding (53). Whether CSE inhibits RhoA via intramolecular

1.0 disulfide bridge formation needs further investigation.
€
∗ ∗ FAK is a key component of FAC and an essential survival
0.5
ε ε factor for anchorage-dependent cells (83). Although FAK ac-
C57BL/6
tivation has been implicated in endothelial barrier dysfunction
AKR
0 in some settings (49, 153), FAK activity is also required for
RA CS RA CS RA CS RA CS 3wks maintenance of endothelial barrier function (72). Endothelial
ctrl LPS ctrl LPS cells isolated from FAK knockout mouse embryos or embryos
expressing kinase-defective FAK displayed increased mono-
B layer permeability (160). Knockdown of FAK by small inter-
AKR fering RNA (siRNA) also caused disruption of tight junctions
€
in the testis (126). FAK activity is also required for resealing of
6
∗
Lung Wet-to-dry

endothelial adhesion junctions after exposure to H2O2 (112)

4 and thrombin (56). We have shown that CSE dose and time
dependently inhibits FAK activity via oxidative stress and that
2 overexpression of FAK attenuates CSE-induced disruption of
AJs, FAC, and F-actin fibers (84). The reactive aldehyde,
0 4-hydroxy-2-nonenal (4-HNE), a component of CSE, also
RA CS RA CS 3wks
decreases FAK activity and endothelial barrier function via
ctrl LPS oxidative stress (137). Taken together, oxidative stress-induced
FAK inhibition contributes to CSE-induced endothelial barrier
C dysfunction.
AKR € Intact microtubules are critical for maintenance of endothe-
(μg/mg lung tissue)

90
lial barrier integrity (17, 91, 139). ␣-Tubulin is a building
Evans blue dye

60 ∗ block of microtubules, and acetylation of ␣-tubulin is charac-

30
0 decreases ␣-tubulin acetylation and causes microtubule depo-
RA CS RA CS 3wks
ctrl LPS
Fig. 3. Effects of prolonged cigarette smoke (CS) exposure on LPS-induced
lung edema. Male 6-wk-old C57BL/6 and AKR mice were exposed to room
air (RA) or CS for 3 wk. One hour after the last CS exposure, mice were
intratracheally administered with 2.5 mg/kg of LPS or equal volume of
saline as a control (ctrl). After 18 h, bronchoalveolar lavage (BAL) protein
content (A), lung wet-to-dry weight ratio (B), and lung extravasation of
albumin-conjugated Evans blue dye (C) were assessed. Three to four
C57BL/6 mice per group and 4 – 6 AKR mice per group were used. εP ⬍
0.05 CS/ctrl vs. RA/ctrl; *P ⬍ 0.05 RA/LPS vs. RA/ctrl; €P ⬍ 0.05
CS/LPS vs. RA/LPS. [Reprinted with permission of the American Journal
of Physiology Lung Cellular and Molecular Physiology (116).]
microvascular endothelial monolayer permeability (120). In
contrast, we have shown that CSE inactivated RhoA via
oxidative stress in a dose- and time-dependent manner (84). In
support of our results, reactive oxygen species (ROS) have
been shown to inhibit RhoA, leading to structural disruption of
FAC and F-actin fibers in other cells (24). We further demon-
strated that overexpression of constitutively active RhoA
blunted CSE-induced disassembly of AJs, FAC, and cortical
F-actin fibers in pulmonary endothelial cells (84). Our data
suggest that inhibition of RhoA contributes to CSE-induced
endothelial barrier dysfunction. The apparent discrepancy of
these results may be due to different preparations and concen-
trations of CSE used. RhoA contains a redox-sensitive motif
[GXXXCGK(S/T)C]. It has been suggested that physiological
levels of ROS and high reduction potential may directly oxi-
teristic of stable microtubules (89, 159). CSE has been shown
to cause microtubule depolymerization in human umbilical
vein endothelial cells (HUVEC; 16). We have shown that CSE
lymerization in pulmonary endothelial cells (20). The micro-
tubule stabilizer paclitaxel elevates ␣-tubulin acetylation and
prevents CSE-induced pulmonary endothelial barrier dysfunc-
tion (20). These results indicate that microtubule instability
contributes to CSE-induced endothelial barrier dysfunction.
Acetylated ␣-tubulin is deacetylated by HDAC6 (158).
HDAC6 is a microtubule-associated protein deacetylase that is
ubiquitously expressed and predominantly located in the cyto-
plasm (58). HDAC6 knockout mice do not have a visible
abnormal phenotype (158) but have better survival and reduced
lung injury after LPS challenge (28, 144). Histone deacetylase
inhibitors attenuate lung injury induced by LPS (96), P. aerugi-
nosa (20), mechanical ventilation (29), and cecal ligation and
puncture (157). HDAC6 inhibitors also blunt increased perme-
ability of lung endothelial monolayers induced by thrombin
(114) and LPS (28). In addition, inhibition of HDAC6 has been
proposed as a novel promising therapeutic option for a variety
of disorders, including cardiac dysfunction, depression, Alz-
heimer’s disease, COPD-associated cilia dysfunction, and di-
abetes (31, 41, 46, 62, 75, 149, 154).
The role of HDAC6 in CS-induced lung injury is less well
understood. We have recently shown that HDAC6 activity is
enhanced in cultured lung endothelial cells exposed to CSE
likely via oxidative stress-mediated Akt inactivation and sub-
sequent GSK-3␤-mediated phosphorylation of HDAC6 at ser-
ine-22 (20). Tubacin, a specific HDAC6 inhibitor (50), and
HDAC6 siRNA abolished CSE-induced barrier dysfunction in
primary human LMVEC in vitro (20). Importantly, tubacin

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 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM L749

EC ET1
Cigarette
smoke
NO

Va
s
oc
SM

on
st
C

ric
pr

tio
ol
Va

ife

n
s
VEGF FAK

ra
od
RhoA FAK

tio
ila

n
AAT UPR

tio
HDAC6 p38

n
Adenosine p53
Ceramide IAM
Ceramide p38
NF-κB
Flow in
remaining
vessels PH in COPD
EC activation EC permeability EC apoptosis
Excessive to
be cleared

Inflammation Lung edema Necrosis

DAMPs
eMPs

Risk for ARDS Autoimmunity

Emphysema
Fig. 4. Proposed model of cigarette smoke-induced pulmonary endothelial cell injury. Cigarette smoke (CS) exposure causes lung endothelial cell (EC) activation,
resulting in inflammation, via activation of NF-␬B signaling; CS exposure directly increases EC permeability likely through multiple signaling pathways, including
inhibition of RhoA, focal adhesion kinase (FAK), and intercellular adhesion molecules (IAM) and activation/upregulation of histone deacetylase-6 (HDAC6), p38, and
ceramide; CS exposure also causes EC apoptosis through signaling pathways involving downregulation of VEGF, FAK, ␣1-antitrypsin (AAT), and unfolded protein
response (UPR) and upregulation of ceramide, adenosine, p38, and p53. Excessive EC apoptosis can lead to necrosis and autoimmunity because of release of
mitochondrial damage-associated molecular patterns (DAMPs), endothelial microparticles (eMPs), and other cellular contents. CS-induced increases in EC activation,
permeability, apoptosis, and necrosis collectively contribute to increased risk of development of acute respiratory distress syndrome (ARDS). CS-induced lung inflammation, EC
apoptosis and necrosis, and autoimmunity cause emphysema. CS exposure increases EC endothelin (ET)-1 levels, leading to increased smooth muscle cell (SMC) proliferation
and vasoconstriction. CS exposure also decreases vasodilation because of reduction in EC NO. All these changes, in combination with increased blood flow in remaining
vessels due to EC apoptosis and loss of vessels, cause vascular remodeling and pulmonary hypertension (PH) in chronic obstructive pulmonary disease (COPD) patients.

prevented CS priming for lung injury after infection with P.
aeruginosa in mice (20). We speculated that HDAC6 inhibitors
may become a novel therapeutic option for CS-associated lung
injury.
Endothelial intercellular adhesion molecule CD146, also
known as cell surface glycoprotein MUC18, is decreased in
lung tissue of smokers with COPD and of rats exposed to CS,
as well as in cultured lung macrovascular and microvascular
endothelial cells exposed to CSE (73). CD146 knockout mice
exhibit perivascular edema and lung inflammation (73). How-
ever, it remains unknown whether loss of CD146 plays a role
in CS-induced endothelial barrier dysfunction and increased
vascular permeability in vivo. Notably, soluble sCD146, which
lacks transmembrane and intracellular domains of CD146, is
increased in plasma and BAL of COPD patients as well as in
BAL of rats exposed to CS (73). Further investigation is
needed to determine whether circulating or BAL sCD146 can
be used as biomarkers of CS-induced lung injury.
Platelet endothelial cell adhesion molecule-1 (PECAM-1)
regulates endothelial barrier permeability by facilitating de-
phosphorylation of ␤-catenin (18). PECAM-1 null mice exhibit
prolonged and increased permeability after inflammatory in-
sults (18). CSE causes a 10-fold increase in tyrosine phosphor-
ylation of PECAM-1 in HUVEC (124). However, the role of
PECAM-1 in CS-induced lung endothelial injury remains un-
known.
It is likely that multiple components of cigarette smoke are
responsible for increased endothelial permeability. We have
demonstrated that acrolein increased endothelial monolayer
and lung microvascular permeability in vivo (82). Schweitzer
et al. reported that nicotine, e-cigarette solution, and condensed
e-cigarette vapor increased endothelial monolayer permeability
(119). In addition, they demonstrated nicotine-independent
effects of e-cigarette solutions on endothelial permeability.
Acrolein is among the components of combustion of e-ciga-
rette solutions.
CS Causes Lung Endothelial Cell Activation
and Inflammation
CS activates the lung endothelium and causes inflammatory
cell accumulation. Sharma et al. showed in human LMVEC
that this mechanism involves the inhibition of platelet aggre-
gating factor acetylhydrolase (PAF-AH), which degrades

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L750 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM
platelet aggregating factor (PAF), leading to increased levels of
PAF (123). Increased PAF is in turn associated with neutrophil
[polymorphonuclear neutrophil (PMN)] adherence to the en-
dothelium. They also showed that CSE stimulates expression
of other endothelium-activating molecules, including P-selec-
tin, E-selectin, intercellular adhesion molecule-1 (ICAM-1),
and vascular cell adhesion molecule-1 (VCAM-1; 123). The
same group subsequently showed that increased PAF is asso-
ciated with increased adherence of metastatic breast cancer
cells to the lung endothelium and that CSE stimulates the
upregulation of the PAF receptor on the breast cancer cells as
well (70).
Acute CS exposure also activates xanthine oxidase and
elicits the rolling and adhesion of leukocytes to endothelium of
arterioles and postcapillary venules of striated muscle micro-
circulation and aortic endothelium in hamsters in vivo; these
effects were prevented by superoxide dismutase and water-
soluble antioxidants but not by lipid-soluble antioxidants (76,
77). CSE increases the surface expression of adhesion mole-
cules, including ICAM-1, endothelial leukocyte adhesion mol-
ecule-1 (ELAM-1), VCAM-1, and E-selectin, as well as cyto-
kines and chemokines, including tumor necrosis factor-␣, IL-6,
and IL-1␤, via NADPH oxidase-dependent NF-␬B transcrip-
tional activation in endothelial cells (98) CSE also increases
adherence of monocytes to the endothelium and transendothe-
lial migration (124), as well as neutrophil transmigration across
HUVEC (99). CS-induced upregulation of the C-X-C motif
chemokine receptor 3 (CXCR3) receptor in endothelial cells
may mediate endothelial cell apoptosis (47). CSE synergizes
with the inflammatory cytokine IL-1␤ to increase vascular
permeability and endothelial dysfunction via ROS/p38/phos-
phatase and tensin homolog (PTEN)-mediated tyrosine phos-
phorylation of vascular endothelial cadherin and ␤-catenin, as
well as subsequent ␤-catenin nuclear translocation and expression
of inflammatory genes, such as cyclooxygenase-2 (COX-2) in
cardiac endothelial cells (11, 12). Taken together, CS causes lung
inflammation via its direct effect on endothelial cell activation,
leading to enhanced endothelial cell apoptosis, increased barrier
permeability, and endothelial dysfunction.
Damage-associated molecular patterns (DAMPs) enhance
PMN adherence to endothelial cells by increasing surface
expression of adhesion molecules in both PMN and human
pulmonary artery endothelial cells in vitro (130). CS exposure
induces necrosis of bronchial epithelial cells and neutrophils,
leading to DAMP release and proinflammatory responses (51,
111). Levels of DAMPs, including high-mobility group box-1
(HMGB1), are increased in extracellular lung fluids of COPD
patients (109), and increased DAMPs have been implicated in
inflammation in COPD (110). Future studies are needed to
address the sources and roles of DAMPs in CS-induced endo-
thelial activation and related lung diseases.
EFFECT OF CIGARETTE SMOKE ON ENDOTHELIAL
APOPTOSIS IN COPD
Robust pulmonary endothelial cell apoptosis has been ob-
served in patients with severe ARDS (1) and in mice with mild
ARDS induced by LPS (40). Pulmonary microvascular endo-
thelial cell apoptosis has been suggested as a cause of barrier
dysfunction and edema in septic mice (44, 45). Inhibition of
apoptosis by a broad-spectrum caspase inhibitor prolonged
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survival of mice exposed to LPS (67). These studies suggest
that lung endothelial cell apoptosis contributes to the patho-
genesis of ARDS.
Recent clinical studies also implicate vascular endothelial
damage in the pathogenesis of COPD. This has been observed
outside the lungs as well, most notably with the loss of
glomerular integrity in patients with COPD. Polverino et al.
(108) showed that COPD patients had lower estimated glomer-
ular filtration rates than smokers without COPD and nonsmok-
ing controls and that lower forced expiratory volume in 1 s
(FEV1) %predicted was associated with double contouring of
the glomerular basement membrane. Most smokers had albu-
minuria as well, but the quantity did not correlate with FEV1.
In mouse studies, cigarette smoke also increased albuminuria.
This group also showed that levels of oxidative stress-ad-
vanced glycation end products (AGEs) and their receptors
(RAGEs) increased in mice with cigarette smoke exposure.
Treatment with enalapril blunted these effects in mice, sug-
gesting a role of angiotensin I-converting enzyme (ACE)
inhibitors for renal protection in patients with COPD (108).
Agrawal et al. also demonstrated increased albuminuria in
patients with acute exacerbations of COPD (3).
Lung tissue from patients with emphysema displays in-
creased apoptosis of alveolar epithelial and endothelial cells
(59, 66). We also observed an increase in lung endothelial cell
apoptosis in AKR mice exposed to CS for 3 wk (115). Circu-
lating endothelial microparticles (eMPs) are thought to be shed
into the bloodstream from activated, apoptotic, or necrotic
endothelial cells. Blood eMPs are significantly elevated in
healthy smokers (128) and patients with COPD (131, 134).
Circulating eMP levels remain elevated in COPD patients
despite smoking cessation but return to normal low levels in
healthy former smokers, suggesting that pulmonary endothelial
cell injury is reversible only in healthy smokers who quit
smoking (128). Circulating eMPs are also increased in rats
exposed to CS for 2– 6 mo (79). Further studies are needed to
determine whether circulating eMPs are markers of vascular
endothelial injury or part of the pathogenesis of diseases linked
to endothelial injury and inflammation in smokers (121).
CSE triggers apoptosis of pulmonary endothelial cells (115,
135) and induces necrosis and inhibits apoptosis of alveolar
epithelial cells and HUVEC (148). The potential mechanisms
underlying CS-induced endothelial apoptosis include inhibi-
tion of VEGF, FAK, and ␣1-antitrypsin (AAT) signaling and
upregulation of ceramide and adenosine, as well as impairment
of unfolded protein response (UPR), as depicted in Fig. 4.
VEGF/VEGFR2 signaling is reduced in the lungs of smok-
ers with and without COPD (59, 66) and of rodents exposed to
CS (88). CSE decreases expression of VEGF in pulmonary
endothelial cells in vitro (135). We observed that FAK activity
is reduced in AKR mice exposed to CS for 3 wk and in lung
endothelial cells exposed to CSE; these effects were associated
with enhanced pulmonary endothelial cell apoptosis in vivo
and in vitro (115). Overexpression of FAK prevented CSE-
induced endothelial cell apoptosis, suggesting that reduced
FAK activity may contribute to CSE-induced endothelial cell
apoptosis (115). Inhibition of FAK also causes emphysema-
like changes in rat lungs (93). Further studies are necessary to
address whether reduced FAK activity contributes to CS-
induced lung endothelial cell apoptosis in vivo. AAT inhibits
apoptosis of cultured lung endothelial cells by blocking inter-
 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM
action of caspases-3, -6, and -7 with their substrates (103).
Overexpression of AAT inhibits apoptosis of cultured lung
endothelial cells and attenuates emphysema caused by active
caspase-3 and blockade of VEGF signaling in vivo (103).
Exogenous AAT protects cultured pulmonary endothelial cells
from CSE-induced apoptosis in vitro (6). Whether exogenous
AAT or overexpression of AAT prevents or reverses CS-
induced lung endothelial cell apoptosis and emphysema in vivo
remains to be determined. Ceramide is upregulated in emphy-
sematous lungs of patients and animal models, as well as in
cultured pulmonary endothelial cells exposed to CSE (104).
This increase in ceramide enhances alveolar cell apoptosis
(104).
CS exposure increases lung tissue adenosine levels in mice,
an effect associated with lung endothelial cell apoptosis and
early emphysema (85). Sustained increased adenosine in aden-
osine deaminase (ADA)-deficient mice enhances alveolar cell
apoptosis and causes emphysema-like changes in mice (161).
ADA expression and activity are reduced in lungs of smokers
with COPD (162). Whether chronically elevated adenosine
contributes to CS-induced lung endothelial cell apoptosis and
development of emphysema remains to be investigated. Other
investigators have reported that CS increased xanthine oxi-
doreductase (XOR) expression and activity and that this in-
crease was sufficient and necessary for p53 induction and
subsequent endothelial cell apoptosis (69).
The UPR is an important mechanism of elimination of
endoplasmic reticulum stress and enhanced cell survival (118).
The UPR is activated in lung tissue of smokers without
emphysema (68). UPR is also activated by CSE in cultured
human bronchial epithelial cells and 3T3 fibroblasts (52, 64)
and in cultured pulmonary endothelial cells (115). Using
mouse models of CS exposure, we have observed a strong link
between impairment of eukaryotic initiation factor 2␣ (eIF2␣)
signaling and lung endothelial cell apoptosis in a mouse strain
susceptible to CS-induced emphysema (115). Future studies
are necessary to determine whether impaired eIF2␣ signaling
contributes to lung endothelial cell apoptosis and emphysema.
In summary, CS exposure causes lung endothelial cell apo-
ptosis. The underlying mechanism is rather complicated, as
summarized in Fig. 4. This may be due to the complexity of
toxins in smoke. Therefore it would be difficult to prevent and
treat cigarette smoke-induced lung diseases by using a single
approach/drug. It may be important to target multiple path-
ways.
EFFECT OF CIGARETTE SMOKE ON ENDOTHELIAL
DYSFUNCTION IN PULMONARY HYPERTENSION
ASSOCIATED WITH COPD
Pulmonary hypertension is a complication of smoking-in-
duced COPD with a prevalence of ~19% in COPD patients
(39). The causes of pulmonary hypertension in COPD are
complex and include vasoconstriction caused by acute hypoxia
and acidosis, loss of microcirculation in emphysema, increased
blood viscosity caused by polycythemia, and vascular remod-
eling caused by chronic hypoxia.
Pulmonary vasoconstriction and vascular remodeling have
been recognized in smokers with normal lung function and in
patients with mild COPD without hypoxemia (117). Pulmo-
nary endothelial dysfunction has also been reported in other-
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L751
wise healthy young smokers (100). In addition, pulmonary
endothelial dysfunction and pulmonary vascular smooth mus-
cle cell proliferation precede emphysema in guinea pigs ex-
posed to cigarette smoke (37). These results suggest that
cigarette smoke directly alters the structure and function of
pulmonary vessels at early stage COPD. CS-induced pulmo-
nary endothelial cell dysfunction might predispose patients
with COPD to further smooth muscle hypertrophy, thus con-
tributing to development of pulmonary hypertension in COPD.
Furthermore, changes in endogenous pulmonary vasoconstric-
tors and vasodilators produced by lung endothelium can cause
vasoconstriction. The potential role of endothelial dysfunction
in the pathophysiology of pulmonary hypertension in COPD
suggests that therapies targeting the endothelium may be ef-
fective in treating these patients.
Endothelin (ET)-1 and nitric oxide (NO) are endothelium-
derived mediators with opposite effects on vascular tone and
cell growth. ET-1 is a potent vasoconstrictor and mitogenic
agent (19). Plasma ET-1 levels are elevated in smokers, and
patients with both COPD and pulmonary hypertension have
increased expression of ET-1 in pulmonary endothelial cells
(156). ET-1 levels are also elevated in hypertensive intrapul-
monary arteries of guinea pigs exposed to CS for 6 mo (151).
CSE promotes ET receptor expression in pulmonary artery
endothelial cells (90). Future studies are needed to address the
role of ET-1 in pulmonary vascular remodeling in COPD
patients with pulmonary hypertension.
NO, a major vasodilator, is produced by endothelial NO
synthase (eNOS). Expression of eNOS is reduced in pulmo-
nary arteries of healthy smokers (10). Pulmonary arterial re-
laxation was also reduced in otherwise healthy smokers; this
effect was associated with decreased eNOS activity and re-
duced NO in exhaled air (34). Diminished lung eNOS levels,
lower NO, and reduced endothelium-dependent vasodilatation
in pulmonary arteries precede the development of emphysema
in guinea pigs (37) and mice (125). After 6 mo of cigarette
smoke exposure, eNOS knockout mice had enhanced vascular
remodeling and pulmonary hypertension (152). CSE irrevers-
ibly decreased eNOS mRNA and protein levels and inhibited
eNOS enzymatic activity in pulmonary endothelial cells (129,
143). CSE also increases eNOS acetylation via oxidative stress-
dependent downregulation of sirtuin 1, leading to blunted eNOS
activity (8). In addition, CSE-induced ROS may interact with NO
to produce peroxynitrite (ONOO⫺), thereby reducing NO bioac-
tivity (61). In fact, reactive nitrogen species are increased in lung
vessels of smokers with COPD and further increased in smok-
ers with combined COPD and pulmonary hypertension (152).
Taken together, oxidative stress-mediated reduction in eNOS-
NO signaling is a significant contributor to CS-induced vascu-
lar endothelial dysfunction.
Nana-Sinkam et al. demonstrated decreased expression of
prostacyclin synthase (PGI2S) and lower levels of prostacyclin,
which prevents apoptosis, in lung tissue of patients with COPD
(95). In cultured human pulmonary microvascular endothelial
cells, treatment with CSE directly decreased expression of
COX-1 and increased expression of COX-2 and cytosolic
phospholipase A2 (CPLA2), potential causes of oxidative
stress. This was prevented by pretreatment with the antioxidant
N-acetylcysteine (95). These results suggest that decreased
prostacyclin and increased oxidative stress also contribute to
L752 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM
CS-induced pulmonary endothelial dysfunction and pulmonary
hypertension in patients with COPD.
PERSPECTIVES AND FUTURE DIRECTIONS
Growing evidence from clinical studies supports an adverse
effect of cigarette smoke exposure as a risk factor for ARDS
and poor outcome. Numerous studies of preclinical animal
models and cultured cells have demonstrated that cigarette
smoke exposure increases pulmonary endothelial barrier per-
meability and causes endothelial activation resulting in en-
hanced lung inflammation. Cigarette smoke exposure also
causes pulmonary endothelial cell apoptosis; this may initiate
and promote development of the lung destruction characteristic
of emphysema. Cigarette smoke exposure directly causes en-
dothelial dysfunction associated with vascular remodeling and
vasoconstriction in smokers.
We hypothesize that the normal alveolar epithelial barrier
protects endothelial cells from exposure to toxins in cigarette
smoke. However, as illustrated in Fig. 4, after smoke-induced
injury to and disruption of the tight alveolar epithelial barrier,
components of smoke and/or DAMPs released from injured
airway epithelial cells can reach and cause injury to endothelial
cells and subsequently enter the circulation via a more perme-
able endothelial barrier. Future studies should include devel-
oping strategies to protect endothelial cells in order to prevent
and treat cigarette smoking-associated lung diseases, such as
ARDS, emphysema, and pulmonary hypertension associated
with COPD. Since components of cigarette smoke and/or
DAMPs released from injured epithelial and endothelial cells
can cross the injured endothelial barrier and enter the systemic
circulation, it would be reasonable to strengthen the pulmonary
endothelial barrier as a measure to prevent cardiovascular and
other cigarette smoke-induced systemic diseases.
GRANTS
This study was supported using facilities at the Providence Veterans Affairs
Medical Center and by Veterans Affairs Merit Review (S. Rounds), National
Heart, Lung, and Blood Institute Grant R01-HL-130230 (Q. Lu), a Brown
University Dean’s Emerging Areas of New Science (DEANS) award (S.
Rounds), National Institute of General Medical Sciences (NIGMS) Grant
U54-GM-115677 (S. Rounds), and an Institutional Development award from
NIGMS under Grant P20-GM-103652 (S. Rounds; project 1 to Q. Lu).
DISCLOSURES
No conflicts of interest, financial or otherwise, are declared by the authors.
AUTHOR CONTRIBUTIONS
Q.L. and E.G. prepared figures; Q.L., E.G., and S.R. drafted manuscript;
Q.L., E.G., and S.R. edited and revised manuscript; Q.L., E.G., and S.R.
approved final version of manuscript.
REFERENCES
1. Abadie Y, Bregeon F, Papazian L, Lange F, Chailley-Heu B, Thomas
P, Duvaldestin P, Adnot S, Maitre B, Delclaux C. Decreased VEGF
concentration in lung tissue and vascular injury during ARDS. Eur
Respir J 25: 139 –146, 2005. doi:10.1183/09031936.04.00065504.
2. Aghajanian A, Wittchen ES, Campbell SL, Burridge K. Direct acti-
vation of RhoA by reactive oxygen species requires a redox-sensitive
motif. PLoS One 4: e8045, 2009. doi:10.1371/journal.pone.0008045.
3. Agrawal A, Garg R, Sahu D, Kumar M. Study the association of
chronic obstructive pulmonary disease with early endothelial dysfunction
and its impact on cardiovascular system by estimating urinary albumin
creatinine ratio. Lung India 34: 138 –143, 2017. doi:10.4103/0970-2113.
201299.
AJP-Lung Cell Mol Physiol • doi:10.1152/ajplung.00373.2017 • www.ajplung.org
Downloaded from journals.physiology.org/journal/ajplung (005.108.050.203) on May 21, 2023.
6. Aldonyte R, Hutchinson TE, Jin B, Brantly M, Block E, Patel J,
Zhang J. Endothelial alpha-1-antitrypsin attenuates cigarette smoke
induced apoptosis in vitro. COPD 5: 153–162, 2008. [Erratum in COPD
5: 405, 2008.] doi:10.1080/15412550802092936.
7. Alwis KU, deCastro BR, Morrow JC, Blount BC. Acrolein exposure
in U.S. tobacco smokers and non-tobacco users: NHANES 2005-2006.
Environ Health Perspect 123: 1302–1308, 2015. doi:10.1289/ehp.
1409251.
8. Arunachalam G, Yao H, Sundar IK, Caito S, Rahman I. SIRT1
regulates oxidant- and cigarette smoke-induced eNOS acetylation in
endothelial cells: role of resveratrol. Biochem Biophys Res Commun 393:
66 –72, 2010. doi:10.1016/j.bbrc.2010.01.080.
9. Barberà JA. Mechanisms of development of chronic obstructive pul-
monary disease-associated pulmonary hypertension. Pulm Circ 3: 160 –
164, 2013. doi:10.4103/2045-8932.109949.
10. Barberà JA, Peinado VI, Santos S, Ramirez J, Roca J, Rodriguez-
Roisin R. Reduced expression of endothelial nitric oxide synthase in
pulmonary arteries of smokers. Am J Respir Crit Care Med 164: 709 –
713, 2001. doi:10.1164/ajrccm.164.4.2101023.
11. Barbieri SS, Ruggiero L, Tremoli E, Weksler BB. Suppressing PTEN
activity by tobacco smoke plus interleukin-1␤ modulates dissociation of
VE-cadherin/␤-catenin complexes in endothelium. Arterioscler Thromb
Vasc Biol 28: 732–738, 2008. doi:10.1161/ATVBAHA.107.159434.
12. Barbieri SS, Weksler BB. Tobacco smoke cooperates with interleu-
kin-1␤ to alter ␤-catenin trafficking in vascular endothelium resulting in
increased permeability and induction of cyclooxygenase-2 expression in
vitro and in vivo. FASEB J 21: 1831–1843, 2007. doi:10.1096/fj.06-
7557com.
13. Barnoya J, Glantz SA. Cardiovascular effects of secondhand smoke:
nearly as large as smoking. Circulation 111: 2684 –2698, 2005. doi:10.
1161/CIRCULATIONAHA.104.492215.
14. Barrowcliffe MP, Jones JG. Pulmonary clearance of 99mTc-DTPA in
the diagnosis and evolution of increased permeability pulmonary oe-
dema. Anaesth Intensive Care 17: 422–432, 1989.
15. Bello S, Menéndez R, Antoni T, Reyes S, Zalacain R, Capelastegui A,
Aspa J, Borderías L, Martin-Villasclaras JJ, Alfageme I, Rodríguez
de Castro F, Rello J, Luis M, Ruiz-Manzano J. Tobacco smoking
increases the risk for death from pneumococcal pneumonia. Chest 146:
1029 –1037, 2014. doi:10.1378/chest.13-2853.
16. Bernhard D, Csordas A, Henderson B, Rossmann A, Kind M, Wick
G. Cigarette smoke metal-catalyzed protein oxidation leads to vascular
endothelial cell contraction by depolymerization of microtubules. FASEB
J 19: 1096 –1107, 2005. doi:10.1096/fj.04-3192com.
17. Birukova AA, Birukov KG, Smurova K, Adyshev D, Kaibuchi K,
Alieva I, Garcia JG, Verin AD. Novel role of microtubules in thrombin-
induced endothelial barrier dysfunction. FASEB J 18: 1879 –1890, 2004.
doi:10.1096/fj.04-2328com.
18. Biswas P, Canosa S, Schoenfeld D, Schoenfeld J, Li P, Cheas LC,
Zhang J, Cordova A, Sumpio B, Madri JA. PECAM-1 affects GSK-
3␤-mediated ␤-catenin phosphorylation and degradation. Am J Pathol
169: 314 –324, 2006. doi:10.2353/ajpath.2006.051112.
19. Böhm F, Pernow J. The importance of endothelin-1 for vascular
dysfunction in cardiovascular disease. Cardiovasc Res 76: 8 –18, 2007.
doi:10.1016/j.cardiores.2007.06.004.
20. Borgas D, Chambers E, Newton J, Ko J, Rivera S, Rounds S, Lu Q.
Cigarette smoke disrupted lung endothelial barrier integrity and increased
susceptibility to acute lung injury via histone deacetylase 6. Am J Respir
Cell Mol Biol 54: 683–696, 2016. doi:10.1165/rcmb.2015-0149OC.
21. Braga VM, Machesky LM, Hall A, Hotchin NA. The small GTPases
Rho and Rac are required for the establishment of cadherin-dependent
cell-cell contacts. J Cell Biol 137: 1421–1431, 1997. doi:10.1083/jcb.
137.6.1421.
22. Brown SM, Wilson E, Presson AP, Zhang C, Dinglas VD, Greene T,
Hopkins RO, Needham DM; National Institutes of Health NHLBI
ARDS Network. Predictors of 6-month health utility outcomes in sur-
vivors of acute respiratory distress syndrome. Thorax 72: 311–317, 2017.
doi:10.1136/thoraxjnl-2016-208560.
23. Burns AR, Hosford SP, Dunn LA, Walker DC, Hogg JC. Respiratory
epithelial permeability after cigarette smoke exposure in guinea pigs. J
Appl Physiol (1985) 66: 2109 –2116, 1989. doi:10.1152/jappl.1989.66.5.
2109.
24. Cai J, Niu X, Chen Y, Hu Q, Shi G, Wu H, Wang J, Yi J.
Emodin-induced generation of reactive oxygen species inhibits RhoA
 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM
activation to sensitize gastric carcinoma cells to anoikis. Neoplasia 10:
41–51, 2008. doi:10.1593/neo.07754.
25. Calfee CS, Delucchi K, Parsons PE, Thompson BT, Ware LB,
Matthay MA; NHLBI ARDS Network. Subphenotypes in acute respi-
ratory distress syndrome: latent class analysis of data from two ran-
domised controlled trials. Lancet Respir Med 2: 611–620, 2014. doi:10.
1016/S2213-2600(14)70097-9.
26. Calfee CS, Matthay MA, Eisner MD, Benowitz N, Call M, Pittet JF,
Cohen MJ. Active and passive cigarette smoking and acute lung injury
after severe blunt trauma. Am J Respir Crit Care Med 183: 1660 –1665,
2011. doi:10.1164/rccm.201011-1802OC.
27. Chan ED, Kinney WH, Honda JR, Bishwakarma R, Gangavelli A,
Mya J, Bai X, Ordway DJ. Tobacco exposure and susceptibility to
tuberculosis: is there a smoking gun? Tuberculosis (Edinb) 94: 544 –550,
2014. doi:10.1016/j.tube.2014.08.010.
28. Chattopadhyay S, Fensterl V, Zhang Y, Veleeparambil M, Wetzel
JL, Sen GC. Inhibition of viral pathogenesis and promotion of the septic
shock response to bacterial infection by IRF-3 are regulated by the
acetylation and phosphorylation of its coactivators. MBio 4: e00636-12,
2013. doi:10.1128/mBio.00636-12.
29. Chen HY, Li L, Fu ZJ. Histone deacetylase inhibitors trichostatin A and
suberoylanilide hydroxamic acid attenuate ventilator-induced lung in-
jury. Pharmazie 69: 55–59, 2014. doi:10.1691/ph.2014.3716.
30. Chun LF, Moazed F, Calfee CS, Matthay MA, Gotts JE. Pulmonary
toxicity of e-cigarettes. Am J Physiol Lung Cell Mol Physiol 313:
L193–L206, 2017. doi:10.1152/ajplung.00071.2017.
31. Demos-Davies KM, Ferguson BS, Cavasin MA, Mahaffey JH, Wil-
liams SM, Spiltoir JI, Schuetze KB, Horn TR, Chen B, Ferrara C,
Scellini B, Piroddi N, Tesi C, Poggesi C, Jeong MY, McKinsey TA.
HDAC6 contributes to pathological responses of heart and skeletal
muscle to chronic angiotensin-II signaling. Am J Physiol Heart Circ
Physiol 307: H252–H258, 2014. doi:10.1152/ajpheart.00149.2014.
32. Divo M. COPD, co-morbidities and health-related quality of life (HRQOL):
more is less. COPD 10: 275–276, 2013. doi:10.3109/15412555.2013.
795409.
33. Drannik AG, Pouladi MA, Robbins CS, Goncharova SI, Kianpour S,
Stämpfli MR. Impact of cigarette smoke on clearance and inflammation
after Pseudomonas aeruginosa infection. Am J Respir Crit Care Med
170: 1164 –1171, 2004. doi:10.1164/rccm.200311-1521OC.
34. Duong-Quy S, Dao P, Hua-Huy T, Guilluy C, Pacaud P, Dinh-Xuan
AT. Increased Rho-kinase expression and activity and pulmonary endo-
thelial dysfunction in smokers with normal lung function. Eur Respir J
37: 349 –355, 2011. doi:10.1183/09031936.00056610.
35. Fabbri LM, Beghé B, Agustí A. COPD and the solar system: introduc-
ing the chronic obstructive pulmonary disease comorbidome. Am J
Respir Crit Care Med 186: 117–119, 2012. doi:10.1164/rccm.201205-
0906ED.
36. Famous KR, Delucchi K, Ware LB, Kangelaris KN, Liu KD, Thomp-
son BT, Calfee CS; ARDS Network. Acute respiratory distress syn-
drome subphenotypes respond differently to randomized fluid manage-
ment strategy. Am J Respir Crit Care Med 195: 331–338, 2017. doi:10.
1164/rccm.201603-0645OC.
37. Ferrer E, Peinado VI, Díez M, Carrasco JL, Musri MM, Martínez A,
Rodríguez-Roisin R, Barberà JA. Effects of cigarette smoke on endo-
thelial function of pulmonary arteries in the guinea pig. Respir Res 10:
76, 2009. doi:10.1186/1465-9921-10-76.
38. Ferro TN, Goslar PW, Romanovsky AA, Petersen SR. Smoking in
trauma patients: the effects on the incidence of sepsis, respiratory failure,
organ failure, and mortality. J Trauma 69: 308 –312, 2010. doi:10.1097/
TA.0b013e3181e1761e.
39. Freixa X, Portillo K, Paré C, Garcia-Aymerich J, Gomez FP, Benet
M, Roca J, Farrero E, Ferrer J, Fernandez-Palomeque C, Antó JM,
Barberà JA; PAC-COPD Study Investigators. Echocardiographic ab-
normalities in patients with COPD at their first hospital admission. Eur
Respir J 41: 784 –791, 2013. doi:10.1183/09031936.00222511.
40. Fujita M, Kuwano K, Kunitake R, Hagimoto N, Miyazaki H, Kaneko
Y, Kawasaki M, Maeyama T, Hara N. Endothelial cell apoptosis in
lipopolysaccharide-induced lung injury in mice. Int Arch Allergy Immu-
nol 117: 202–208, 1998. doi:10.1159/000024011.
41. Fukada M, Hanai A, Nakayama A, Suzuki T, Miyata N, Rodriguiz
RM, Wetsel WC, Yao TP, Kawaguchi Y. Loss of deacetylation activity
of Hdac6 affects emotional behavior in mice. PLoS One 7: e30924, 2012.
doi:10.1371/journal.pone.0030924.
AJP-Lung Cell Mol Physiol • doi:10.1152/ajplung.00373.2017 • www.ajplung.org
Downloaded from journals.physiology.org/journal/ajplung (005.108.050.203) on May 21, 2023.
L753
42. Gajic O, Dabbagh O, Park PK, Adesanya A, Chang SY, Hou P,
Anderson H III, Hoth JJ, Mikkelsen ME, Gentile NT, Gong MN,
Talmor D, Bajwa E, Watkins TR, Festic E, Yilmaz M, Iscimen R,
Kaufman DA, Esper AM, Sadikot R, Douglas I, Sevransky J, Ma-
linchoc M; U.S. Critical Illness and Injury Trials Group: Lung
Injury Prevention Study Investigators (USCIITG-LIPS). Early iden-
tification of patients at risk of acute lung injury: evaluation of lung injury
prediction score in a multicenter cohort study. Am J Respir Crit Care
Med 183: 462–470, 2011. doi:10.1164/rccm.201004-0549OC.
43. García-Lucio J, Argemi G, Tura-Ceide O, Diez M, Paul T, Bonjoch
C, Coll-Bonfill N, Blanco I, Barberà JA, Musri MM, Peinado VI.
Gene expression profile of angiogenic factors in pulmonary arteries in
COPD: relationship with vascular remodeling. Am J Physiol Lung Cell
Mol Physiol 310: L583–L592, 2016. doi:10.1152/ajplung.00261.2015.
44. Gill SE, Rohan M, Mehta S. Role of pulmonary microvascular endo-
thelial cell apoptosis in murine sepsis-induced lung injury in vivo. Respir
Res 16: 109, 2015. doi:10.1186/s12931-015-0266-7.
45. Gill SE, Taneja R, Rohan M, Wang L, Mehta S. Pulmonary micro-
vascular albumin leak is associated with endothelial cell death in murine
sepsis-induced lung injury in vivo. PLoS One 9: e88501, 2014. doi:10.
1371/journal.pone.0088501.
46. Govindarajan N, Rao P, Burkhardt S, Sananbenesi F, Schlüter OM,
Bradke F, Lu J, Fischer A. Reducing HDAC6 ameliorates cognitive
deficits in a mouse model for Alzheimer’s disease. EMBO Mol Med 5:
52–63, 2013. doi:10.1002/emmm.201201923.
47. Green LA, Petrusca D, Rajashekhar G, Gianaris T, Schweitzer KS,
Wang L, Justice MJ, Petrache I, Clauss M. Cigarette smoke-induced
CXCR3 receptor up-regulation mediates endothelial apoptosis. Am J
Respir Cell Mol Biol 47: 807–814, 2012. doi:10.1165/rcmb.2012-
0132OC.
48. Guerassimov A, Hoshino Y, Takubo Y, Turcotte A, Yamamoto M,
Ghezzo H, Triantafillopoulos A, Whittaker K, Hoidal JR, Cosio MG.
The development of emphysema in cigarette smoke-exposed mice is
strain dependent. Am J Respir Crit Care Med 170: 974 –980, 2004.
doi:10.1164/rccm.200309-1270OC.
49. Guo M, Wu MH, Granger HJ, Yuan SY. Focal adhesion kinase in
neutrophil-induced microvascular hyperpermeability. Microcirculation
12: 223–232, 2005. doi:10.1080/10739680590905251.
50. Haggarty SJ, Koeller KM, Wong JC, Grozinger CM, Schreiber SL.
Domain-selective small-molecule inhibitor of histone deacetylase 6
(HDAC6)-mediated tubulin deacetylation. Proc Natl Acad Sci USA 100:
4389 –4394, 2003. doi:10.1073/pnas.0430973100.
51. Heijink IH, Pouwels SD, Leijendekker C, de Bruin HG, Zijlstra GJ,
van der Vaart H, Ten Hacken NH, van Oosterhout AJ, Nawijn MC,
van der Toorn M. Cigarette smoke induced damage-associated molec-
ular pattern release from necrotic neutrophils triggers proinflammatory
mediator release. Am J Respir Cell Mol Biol 52: 554 –562, 2015.
doi:10.1165/rcmb.2013-0505OC.
52. Hengstermann A, Müller T. Endoplasmic reticulum stress induced by
aqueous extracts of cigarette smoke in 3T3 cells activates the unfolded-
protein-response-dependent PERK pathway of cell survival. Free Radic
Biol Med 44: 1097–1107, 2008. doi:10.1016/j.freeradbiomed.2007.12.
009.
53. Heo J, Raines KW, Mocanu V, Campbell SL. Redox regulation of
RhoA. Biochemistry 45: 14481–14489, 2006. doi:10.1021/bi0610101.
54. Hirsch J, Chalkley RJ, Bentley T, Burlingame AL, Frank JA. Double
impact of cigarette smoke and mechanical ventilation on the alveolar
epithelial type II cell. Crit Care 18: R50, 2014. doi:10.1186/cc13795.
55. Holden WE, Maier JM, Malinow MR. Cigarette smoke extract in-
creases albumin flux across pulmonary endothelium in vitro. J Appl
Physiol (1985) 66: 443–449, 1989. doi:10.1152/jappl.1989.66.1.443.
56. Holinstat M, Knezevic N, Broman M, Samarel AM, Malik AB,
Mehta D. Suppression of RhoA activity by focal adhesion kinase-
induced activation of p190RhoGAP: role in regulation of endothelial
permeability. J Biol Chem 281: 2296 –2305, 2006. doi:10.1074/jbc.
M511248200.
57. Hsieh SJ, Zhuo H, Benowitz NL, Thompson BT, Liu KD, Matthay
MA, Calfee CS; National Heart, Lung, and Blood Institute Acute
Respiratory Distress Syndrome Network; National Heart Lung and
Blood Institute Acute Respiratory Distress Syndrome Network. Prev-
alence and impact of active and passive cigarette smoking in acute
respiratory distress syndrome. Crit Care Med 42: 2058 –2068, 2014.
doi:10.1097/CCM.0000000000000418.
L754 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM
58. Hubbert C, Guardiola A, Shao R, Kawaguchi Y, Ito A, Nixon A,
Yoshida M, Wang XF, Yao TP. HDAC6 is a microtubule-associated
deacetylase. Nature 417: 455–458, 2002. doi:10.1038/417455a.
59. Imai K, Mercer BA, Schulman LL, Sonett JR, D’Armiento JM.
Correlation of lung surface area to apoptosis and proliferation in human
emphysema. Eur Respir J 25: 250 –258, 2005. doi:10.1183/09031936.
05.00023704.
60. Iribarren C, Jacobs DR Jr, Sidney S, Gross MD, Eisner MD.
Cigarette smoking, alcohol consumption, and risk of ARDS: a 15-year
cohort study in a managed care setting. Chest 117: 163–168, 2000.
doi:10.1378/chest.117.1.163.
61. Jaimes EA, DeMaster EG, Tian RX, Raij L. Stable compounds of
cigarette smoke induce endothelial superoxide anion production via
NADPH oxidase activation. Arterioscler Thromb Vasc Biol 24: 1031–
1036, 2004. doi:10.1161/01.ATV.0000127083.88549.58.
62. Jochems J, Boulden J, Lee BG, Blendy JA, Jarpe M, Mazitschek R,
Van Duzer JH, Jones S, Berton O. Antidepressant-like properties of
novel HDAC6-selective inhibitors with improved brain bioavailability.
Neuropsychopharmacology 39: 389 –400, 2014. doi:10.1038/npp.2013.
207.
63. Jones JG, Lawler P, Crawley JC, Minty BD, Hulands G, Veall N.
Increased alveolar epithelial permeability in cigarette smokers. Lancet
315: 66 –68, 1980. doi:10.1016/S0140-6736(80)90493-6.
64. Jorgensen E, Stinson A, Shan L, Yang J, Gietl D, Albino AP.
Cigarette smoke induces endoplasmic reticulum stress and the unfolded
protein response in normal and malignant human lung cells. BMC Cancer
8: 229, 2008. doi:10.1186/1471-2407-8-229.
65. Kang MJ, Lee CG, Lee JY, Dela Cruz CS, Chen ZJ, Enelow R, Elias
JA. Cigarette smoke selectively enhances viral PAMP- and virus-in-
duced pulmonary innate immune and remodeling responses in mice. J
Clin Invest 118: 2771–2784, 2008. doi:10.1172/JCI32709.
66. Kasahara Y, Tuder RM, Cool CD, Lynch DA, Flores SC, Voelkel NF.
Endothelial cell death and decreased expression of vascular endothelial
growth factor and vascular endothelial growth factor receptor 2 in
emphysema. Am J Respir Crit Care Med 163: 737–744, 2001. doi:10.
1164/ajrccm.163.3.2002117.
67. Kawasaki M, Kuwano K, Hagimoto N, Matsuba T, Kunitake R,
Tanaka T, Maeyama T, Hara N. Protection from lethal apoptosis in
lipopolysaccharide-induced acute lung injury in mice by a caspase
inhibitor. Am J Pathol 157: 597–603, 2000. doi:10.1016/S0002-9440(10)
64570-1.
68. Kelsen SG, Duan X, Ji R, Perez O, Liu C, Merali S. Cigarette smoke
induces an unfolded protein response in the human lung: a proteomic
approach. Am J Respir Cell Mol Biol 38: 541–550, 2008. doi:10.1165/
rcmb.2007-0221OC.
69. Kim BS, Serebreni L, Hamdan O, Wang L, Parniani A, Sussan T,
Scott Stephens R, Boyer L, Damarla M, Hassoun PM, Damico R.
Xanthine oxidoreductase is a critical mediator of cigarette smoke-in-
duced endothelial cell DNA damage and apoptosis. Free Radic Biol Med
60: 336 –346, 2013. doi:10.1016/j.freeradbiomed.2013.01.023.
70. Kispert SE, Marentette JO, McHowat J. Enhanced breast cancer cell
adherence to the lung endothelium via PAF acetylhydrolase inhibition: a
potential mechanism for enhanced metastasis in smokers. Am J Physiol
Cell Physiol 307: C951–C956, 2014. doi:10.1152/ajpcell.00218.2014.
71. Kitaguchi Y, Taraseviciene-Stewart L, Hanaoka M, Natarajan R,
Kraskauskas D, Voelkel NF. Acrolein induces endoplasmic reticulum
stress and causes airspace enlargement. PLoS One 7: e38038, 2012.
doi:10.1371/journal.pone.0038038.
72. Knezevic N, Tauseef M, Thennes T, Mehta D. The G protein ␤␥
subunit mediates reannealing of adherens junctions to reverse endothelial
permeability increase by thrombin. J Exp Med 206: 2761–2777, 2009.
doi:10.1084/jem.20090652.
73. Kratzer A, Chu HW, Salys J, Moumen Z, Leberl M, Bowler R, Cool
C, Zamora M, Taraseviciene-Stewart L. Endothelial cell adhesion
molecule CD146: implications for its role in the pathogenesis of COPD.
J Pathol 230: 388 –398, 2013. doi:10.1002/path.4197.
74. Kubo S, Kobayashi M, Masunaga Y, Ishii H, Hirano Y, Takahashi K,
Shimizu Y. Cytokine and chemokine expression in cigarette smoke-
induced lung injury in guinea pigs. Eur Respir J 26: 993–1001, 2005.
doi:10.1183/09031936.05.00042405.
75. Lam HC, Cloonan SM, Bhashyam AR, Haspel JA, Singh A, Sathi-
rapongsasuti JF, Cervo M, Yao H, Chung AL, Mizumura K, An CH,
Shan B, Franks JM, Haley KJ, Owen CA, Tesfaigzi Y, Washko GR,
Quackenbush J, Silverman EK, Rahman I, Kim HP, Mahmood A,
AJP-Lung Cell Mol Physiol • doi:10.1152/ajplung.00373.2017 • www.ajplung.org
Downloaded from journals.physiology.org/journal/ajplung (005.108.050.203) on May 21, 2023.
Biswal SS, Ryter SW, Choi AM. Histone deacetylase 6-mediated
selective autophagy regulates COPD-associated cilia dysfunction. J Clin
Invest 123: 5212–5230, 2013. doi:10.1172/JCI69636.
76. Lehr HA, Frei B, Arfors KE. Vitamin C prevents cigarette smoke-
induced leukocyte aggregation and adhesion to endothelium in vivo.
Proc Natl Acad Sci USA 91: 7688 –7692, 1994. doi:10.1073/pnas.91.16.
7688.
77. Lehr HA, Kress E, Menger MD, Friedl HP, Hübner C, Arfors KE,
Messmer K. Cigarette smoke elicits leukocyte adhesion to endothelium
in hamsters: inhibition by CuZn-SOD. Free Radic Biol Med 14: 573–581,
1993. doi:10.1016/0891-5849(93)90138-K.
78. Li XY, Rahman I, Donaldson K, MacNee W. Mechanisms of cigarette
smoke induced increased airspace permeability. Thorax 51: 465–471,
1996. doi:10.1136/thx.51.5.465.
79. Liu H, Ding L, Zhang Y, Ni S. Circulating endothelial microparticles
involved in lung function decline in a rat exposed in cigarette smoke
maybe from apoptotic pulmonary capillary endothelial cells. J Thorac
Dis 6: 649 –655, 2014. doi:10.3978/j.issn.2072-1439.2014.06.26.
80. Liu X, Liu Y, Huang X, Lin G, Xie C. Endothelial progenitor cell
dysfunction in acute exacerbation of chronic obstructive pulmonary
disease. Mol Med Rep 16: 5294 –5302, 2017. doi:10.3892/mmr.2017.
7260.
81. Low B, Liang M, Fu J. p38 mitogen-activated protein kinase mediates
sidestream cigarette smoke-induced endothelial permeability. J Pharma-
col Sci 104: 225–231, 2007. doi:10.1254/jphs.FP0070385.
82. Lu Q, Mundy M, Chambers E, Lange T, Newton J, Borgas D, Yao H,
Choudhary G, Basak R, Oldham M, Rounds S. Alda-1 protects against
acrolein-induced acute lung injury and endothelial barrier dysfunction.
Am J Respir Cell Mol Biol 57: 662–673, 2017. doi:10.1165/rcmb.2016-
0342OC.
83. Lu Q, Rounds S. Focal adhesion kinase and endothelial cell apoptosis.
Microvasc Res 83: 56 –63, 2012. doi:10.1016/j.mvr.2011.05.003.
84. Lu Q, Sakhatskyy P, Grinnell K, Newton J, Ortiz M, Wang Y,
Sanchez-Esteban J, Harrington EO, Rounds S. Cigarette smoke
causes lung vascular barrier dysfunction via oxidative stress-mediated
inhibition of RhoA and focal adhesion kinase. Am J Physiol Lung Cell
Mol Physiol 301: L847–L857, 2011. doi:10.1152/ajplung.00178.2011.
85. Lu Q, Sakhatskyy P, Newton J, Shamirian P, Hsiao V, Curren S,
Gabino Miranda GA, Pedroza M, Blackburn MR, Rounds S. Sus-
tained adenosine exposure causes lung endothelial apoptosis: a possible
contributor to cigarette smoke-induced endothelial apoptosis and lung
injury. Am J Physiol Lung Cell Mol Physiol 304: L361–L370, 2013.
doi:10.1152/ajplung.00161.2012.
86. Lugade AA, Bogner PN, Thatcher TH, Sime PJ, Phipps RP,
Thanavala Y. Cigarette smoke exposure exacerbates lung inflammation
and compromises immunity to bacterial infection. J Immunol 192:
5226 –5235, 2014. doi:10.4049/jimmunol.1302584.
88. Marwick JA, Stevenson CS, Giddings J, MacNee W, Butler K,
Rahman I, Kirkham PA. Cigarette smoke disrupts VEGF165-
VEGFR-2 receptor signaling complex in rat lungs and patients with
COPD: morphological impact of VEGFR-2 inhibition. Am J Physiol
Lung Cell Mol Physiol 290: L897–L908, 2006. doi:10.1152/ajplung.
00116.2005.
89. Matsuyama A, Shimazu T, Sumida Y, Saito A, Yoshimatsu Y,
Seigneurin-Berny D, Osada H, Komatsu Y, Nishino N, Khochbin S,
Horinouchi S, Yoshida M. In vivo destabilization of dynamic microtu-
bules by HDAC6-mediated deacetylation. EMBO J 21: 6820 –6831,
2002. doi:10.1093/emboj/cdf682.
90. Milara J, Ortiz JL, Juan G, Guijarro R, Almudever P, Martorell M,
Morcillo EJ, Cortijo J. Cigarette smoke exposure up-regulates endo-
thelin receptor B in human pulmonary artery endothelial cells: molecular
and functional consequences. Br J Pharmacol 161: 1599 –1615, 2010.
doi:10.1111/j.1476-5381.2010.00979.x.
91. Mirzapoiazova T, Kolosova IA, Moreno L, Sammani S, Garcia JG,
Verin AD. Suppression of endotoxin-induced inflammation by taxol. Eur
Respir J 30: 429 –435, 2007. doi:10.1183/09031936.00154206.
92. Mizumura K, Cloonan SM, Nakahira K, Bhashyam AR, Cervo M,
Kitada T, Glass K, Owen CA, Mahmood A, Washko GR, Hashimoto
S, Ryter SW, Choi AM. Mitophagy-dependent necroptosis contributes
to the pathogenesis of COPD. J Clin Invest 124: 3987–4003, 2014.
doi:10.1172/JCI74985.
93. Mizuno S, Yasuo M, Bogaard HJ, Kraskauskas D, Alhussaini A,
Gomez-Arroyo J, Farkas D, Farkas L, Voelkel NF. Copper deficiency
 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM
induced emphysema is associated with focal adhesion kinase inactiva-
tion. PLoS One 7: e30678, 2012. doi:10.1371/journal.pone.0030678.
94. Moazed F, Burnham EL, Vandivier RW, O’Kane CM, Shyamsundar
M, Hamid U, Abbott J, Thickett DR, Matthay MA, McAuley DF,
Calfee CS. Cigarette smokers have exaggerated alveolar barrier disrup-
tion in response to lipopolysaccharide inhalation. Thorax 71: 1130 –1136,
2016. doi:10.1136/thoraxjnl-2015-207886.
95. Nana-Sinkam SP, Lee JD, Sotto-Santiago S, Stearman RS, Keith RL,
Choudhury Q, Cool C, Parr J, Moore MD, Bull TM, Voelkel NF,
Geraci MW. Prostacyclin prevents pulmonary endothelial cell apoptosis
induced by cigarette smoke. Am J Respir Crit Care Med 175: 676 –685,
2007. doi:10.1164/rccm.200605-724OC.
96. Ni YF, Wang J, Yan XL, Tian F, Zhao JB, Wang YJ, Jiang T.
Histone deacetylase inhibitor, butyrate, attenuates lipopolysaccharide-
induced acute lung injury in mice. Respir Res 11: 33, 2010. doi:10.1186/
1465-9921-11-33.
98. Orosz Z, Csiszar A, Labinskyy N, Smith K, Kaminski PM, Ferdi-
nandy P, Wolin MS, Rivera A, Ungvari Z. Cigarette smoke-induced
proinflammatory alterations in the endothelial phenotype: role of NA-
D(P)H oxidase activation. Am J Physiol Heart Circ Physiol 292: H130 –
H139, 2007. doi:10.1152/ajpheart.00599.2006.
99. Overbeek SA, Braber S, Henricks PA, Kleinjan M, Kamp VM,
Georgiou NA, Garssen J, Kraneveld AD, Folkerts G. Cigarette smoke
induces ␤2-integrin-dependent neutrophil migration across human endo-
thelium. Respir Res 12: 75, 2011. doi:10.1186/1465-9921-12-75.
100. Ozaki K, Hori T, Ishibashi T, Nishio M, Aizawa Y. Effects of chronic
cigarette smoking on endothelial function in young men. J Cardiol 56:
307–313, 2010. doi:10.1016/j.jjcc.2010.07.003.
101. Panzer AR, Lynch SV, Langelier C, Christie JD, McCauley K,
Nelson M, Cheung CK, Benowitz NL, Cohen MJ, Calfee CS. Lung
microbiota is related to smoking status and to development of acute
respiratory distress syndrome in critically ill trauma patients. Am J Respir
Crit Care Med 197: 621–631, 2018. doi:10.1164/rccm.201702-0441OC.
102. Petrache I, Birukova A, Ramirez SI, Garcia JG, Verin AD. The role
of the microtubules in tumor necrosis factor-␣-induced endothelial cell
permeability. Am J Respir Cell Mol Biol 28: 574 –581, 2003. doi:10.
1165/rcmb.2002-0075OC.
103. Petrache I, Fijalkowska I, Medler TR, Skirball J, Cruz P, Zhen L,
Petrache HI, Flotte TR, Tuder RM. ␣-1 antitrypsin inhibits caspase-3
activity, preventing lung endothelial cell apoptosis. Am J Pathol 169:
1155–1166, 2006. doi:10.2353/ajpath.2006.060058.
104. Petrache I, Natarajan V, Zhen L, Medler TR, Richter AT, Cho C,
Hubbard WC, Berdyshev EV, Tuder RM. Ceramide upregulation
causes pulmonary cell apoptosis and emphysema-like disease in mice.
Nat Med 11: 491–498, 2005. doi:10.1038/nm1238.
105. Petta V, Bakakos P, Tseliou E, Kostikas K, Simoes DC, Konstantel-
lou E, Hillas G, Koulouris NG, Papiris S, Loukides S. Angiopoietins
1 and 2 in sputum supernatant of optimally treated asthmatics: the effect
of smoking. Eur J Clin Invest 45: 56 –62, 2015. doi:10.1111/eci.12379.
106. Phillips E, Wang TW, Husten CG, Corey CG, Apelberg BJ, Jamal A,
Homa DM, King BA. Tobacco product use among adults: United States,
2015. MMWR Morb Mortal Wkly Rep 66: 1209 –1215, 2017. doi:10.
15585/mmwr.mm6644a2.
107. Pines H, Koutsky L, Buskin S. Cigarette smoking and mortality among
HIV-infected individuals in Seattle, Washington (1996 –2008). AIDS
Behav 15: 243–251, 2011. doi:10.1007/s10461-010-9682-3.
108. Polverino F, Laucho-Contreras ME, Petersen H, Bijol V, Sholl LM,
Choi ME, Divo M, Pinto-Plata V, Chetta A, Tesfaigzi Y, Celli BR,
Owen CA. A pilot study linking endothelial injury in lungs and kidneys
in chronic obstructive pulmonary disease. Am J Respir Crit Care Med
195: 1464 –1476, 2017. doi:10.1164/rccm.201609-1765OC.
109. Pouwels SD, Heijink IH, ten Hacken NH, Vandenabeele P, Krysko
DV, Nawijn MC, van Oosterhout AJ. DAMPs activating innate and
adaptive immune responses in COPD. Mucosal Immunol 7: 215–226,
2014. doi:10.1038/mi.2013.77.
110. Pouwels SD, Hesse L, Faiz A, Lubbers J, Bodha PK, Ten Hacken
NH, van Oosterhout AJ, Nawijn MC, Heijink IH. Susceptibility for
cigarette smoke-induced DAMP release and DAMP-induced inflamma-
tion in COPD. Am J Physiol Lung Cell Mol Physiol 311: L881–L892,
2016. doi:10.1152/ajplung.00135.2016.
111. Pouwels SD, Zijlstra GJ, van der Toorn M, Hesse L, Gras R, Ten
Hacken NH, Krysko DV, Vandenabeele P, de Vries M, van Ooster-
hout AJ, Heijink IH, Nawijn MC. Cigarette smoke-induced necroptosis
and DAMP release trigger neutrophilic airway inflammation in mice. Am
AJP-Lung Cell Mol Physiol • doi:10.1152/ajplung.00373.2017 • www.ajplung.org
Downloaded from journals.physiology.org/journal/ajplung (005.108.050.203) on May 21, 2023.
L755
J Physiol Lung Cell Mol Physiol 310: L377–L386, 2016. doi:10.1152/
ajplung.00174.2015.
112. Quadri SK, Bhattacharya J. Resealing of endothelial junctions by focal
adhesion kinase. Am J Physiol Lung Cell Mol Physiol 292: L334 –L342,
2007. doi:10.1152/ajplung.00228.2006.
113. Reddy KP, Parker RA, Losina E, Baggett TP, Paltiel AD, Rigotti NA,
Weinstein MC, Freedberg KA, Walensky RP. Impact of cigarette
smoking and smoking cessation on life expectancy among people with
HIV: a US-based modeling study. J Infect Dis 214: 1672–1681, 2016.
doi:10.1093/infdis/jiw430.
114. Saito S, Lasky JA, Guo W, Nguyen H, Mai A, Danchuk S, Sullivan
DE, Shan B. Pharmacological inhibition of HDAC6 attenuates endothe-
lial barrier dysfunction induced by thrombin. Biochem Biophys Res
Commun 408: 630 –634, 2011. doi:10.1016/j.bbrc.2011.04.075.
115. Sakhatskyy P, Gabino Miranda GA, Newton J, Lee CG, Choudhary
G, Vang A, Rounds S, Lu Q. Cigarette smoke-induced lung endothelial
apoptosis and emphysema are associated with impairment of FAK and
eIF2␣. Microvasc Res 94: 80 –89, 2014. doi:10.1016/j.mvr.2014.05.003.
116. Sakhatskyy P, Wang Z, Borgas D, Lomas-Neira J, Chen Y, Ayala A,
Rounds S, Lu Q. Double-hit mouse model of cigarette smoke priming
for acute lung injury. Am J Physiol Lung Cell Mol Physiol 312: L56 –
L67, 2017. doi:10.1152/ajplung.00436.2016.
117. Santos S, Peinado VI, Ramírez J, Melgosa T, Roca J, Rodriguez-
Roisin R, Barberà JA. Characterization of pulmonary vascular remod-
elling in smokers and patients with mild COPD. Eur Respir J 19:
632–638, 2002. doi:10.1183/09031936.02.00245902.
118. Schröder M, Kaufman RJ. The mammalian unfolded protein response.
Annu Rev Biochem 74: 739 –789, 2005. doi:10.1146/annurev.biochem.
73.011303.074134.
119. Schweitzer KS, Chen SX, Law S, Van Demark M, Poirier C, Justice
MJ, Hubbard WC, Kim ES, Lai X, Wang M, Kranz WD, Carroll CJ,
Ray BD, Bittman R, Goodpaster J, Petrache I. Endothelial disruptive
proinflammatory effects of nicotine and e-cigarette vapor exposures. Am
J Physiol Lung Cell Mol Physiol 309: L175–L187, 2015. doi:10.1152/
ajplung.00411.2014.
120. Schweitzer KS, Hatoum H, Brown MB, Gupta M, Justice MJ, Beteck
B, Van Demark M, Gu Y, Presson RG Jr, Hubbard WC, Petrache I.
Mechanisms of lung endothelial barrier disruption induced by cigarette
smoke: role of oxidative stress and ceramides. Am J Physiol Lung Cell
Mol Physiol 301: L836 –L846, 2011. doi:10.1152/ajplung.00385.2010.
121. Serban KA, Rezania S, Petrusca DN, Poirier C, Cao D, Justice MJ,
Patel M, Tsvetkova I, Kamocki K, Mikosz A, Schweitzer KS, Jacob-
son S, Cardoso A, Carlesso N, Hubbard WC, Kechris K, Dragnea B,
Berdyshev EV, McClintock J, Petrache I. Structural and functional
characterization of endothelial microparticles released by cigarette
smoke. Sci Rep 6: 31596, 2016. doi:10.1038/srep31596.
122. Shankar-Hari M, McAuley DF. Acute respiratory distress syndrome
phenotypes and identifying treatable traits. The dawn of personalized
medicine for ARDS. Am J Respir Crit Care Med 195: 280 –281, 2017.
doi:10.1164/rccm.201608-1729ED.
123. Sharma J, Young DM, Marentette JO, Rastogi P, Turk J, McHowat
J. Lung endothelial cell platelet-activating factor production and inflam-
matory cell adherence are increased in response to cigarette smoke
component exposure. Am J Physiol Lung Cell Mol Physiol 302: L47–
L55, 2012. doi:10.1152/ajplung.00179.2011.
124. Shen Y, Rattan V, Sultana C, Kalra VK. Cigarette smoke condensate-
induced adhesion molecule expression and transendothelial migration of
monocytes. Am J Physiol Heart Circ Physiol 270: H1624 –H1633, 1996.
doi:10.1152/ajpheart.1996.270.5.H1624.
125. Sikka G, Pandey D, Bhuniya AK, Steppan J, Armstrong D, Santha-
nam L, Nyhan D, Berkowitz DE. Contribution of arginase activation to
vascular dysfunction in cigarette smoking. Atherosclerosis 231: 91–94,
2013. doi:10.1016/j.atherosclerosis.2013.08.026.
126. Siu ER, Wong EW, Mruk DD, Porto CS, Cheng CY. Focal adhesion
kinase is a blood-testis barrier regulator. Proc Natl Acad Sci USA 106:
9298 –9303, 2009. doi:10.1073/pnas.0813113106.
127. Stämpfli MR, Anderson GP. How cigarette smoke skews immune
responses to promote infection, lung disease and cancer. Nat Rev Immu-
nol 9: 377–384, 2009. doi:10.1038/nri2530.
128. Strulovici-Barel Y, Staudt MR, Krause A, Gordon C, Tilley AE,
Harvey BG, Kaner RJ, Hollmann C, Mezey JG, Bitter H, Pillai SG,
Hilton H, Wolff G, Stevenson CS, Visvanathan S, Fine JS, Crystal
RG. Persistence of circulating endothelial microparticles in COPD de-
L756 EFFECTS OF CIGARETTE SMOKE ON PULMONARY ENDOTHELIUM
spite smoking cessation. Thorax 71: 1137–1144, 2016. doi:10.1136/
thoraxjnl-2015-208274.
129. Su Y, Han W, Giraldo C, De Li Y, Block ER. Effect of cigarette smoke
extract on nitric oxide synthase in pulmonary artery endothelial cells. Am J
Respir Cell Mol Biol 19: 819 –825, 1998. doi:10.1165/ajrcmb.19.5.3091.
130. Sun S, Sursal T, Adibnia Y, Zhao C, Zheng Y, Li H, Otterbein LE,
Hauser CJ, Itagaki K. Mitochondrial DAMPs increase endothelial
permeability through neutrophil dependent and independent pathways.
PLoS One 8: e59989, 2013. doi:10.1371/journal.pone.0059989.
131. Takahashi T, Kobayashi S, Fujino N, Suzuki T, Ota C, He M,
Yamada M, Suzuki S, Yanai M, Kurosawa S, Yamaya M, Kubo H.
Increased circulating endothelial microparticles in COPD patients: a
potential biomarker for COPD exacerbation susceptibility. Thorax 67:
1067–1074, 2012. doi:10.1136/thoraxjnl-2011-201395.
132. Tandon S, Batchelor A, Bullock R, Gascoigne A, Griffin M, Hayes N,
Hing J, Shaw I, Warnell I, Baudouin SV. Peri-operative risk factors for
acute lung injury after elective oesophagectomy. Br J Anaesth 86:
633–638, 2001. doi:10.1093/bja/86.5.633.
133. Taylor AE, Gaar KA Jr. Estimation of equivalent pore radii of pulmo-
nary capillary and alveolar membranes. Am J Physiol 218: 1133–1140,
1970. doi:10.1152/ajplegacy.1970.218.4.1133.
134. Thomashow MA, Shimbo D, Parikh MA, Hoffman EA, Vogel-Claussen
J, Hueper K, Fu J, Liu CY, Bluemke DA, Ventetuolo CE, Doyle MF,
Barr RG. Endothelial microparticles in mild chronic obstructive pulmonary
disease and emphysema. The Multi-Ethnic Study of Atherosclerosis Chronic
Obstructive Pulmonary Disease study. Am J Respir Crit Care Med 188:
60 –68, 2013. doi:10.1164/rccm.201209-1697OC.
135. Tuder RM, Wood K, Taraseviciene L, Flores SC, Voekel NF. Ciga-
rette smoke extract decreases the expression of vascular endothelial
growth factor by cultured cells and triggers apoptosis of pulmonary
endothelial cells. Chest 117, Suppl 1: 241S–242S, 2000. doi:10.1378/
chest.117.5_suppl_1.241S.
136. Tzima E. Role of small GTPases in endothelial cytoskeletal dynamics
and the shear stress response. Circ Res 98: 176 –185, 2006. doi:10.1161/
01.RES.0000200162.94463.d7.
136a.United States. Department of Health and Human Services. The
Health Consequences of Smoking: 50 Years of Progress. A Report of the
Surgeon General. Atlanta, GA: U.S. Dept. of Health and Human Ser-
vices, Centers for Disease Control and Prevention, National Center for
Chronic Disease Prevention and Health Promotion, Office on Smoking
and Health, 2014.
137. Usatyuk PV, Parinandi NL, Natarajan V. Redox regulation of 4-hy-
droxy-2-nonenal-mediated endothelial barrier dysfunction by focal ad-
hesion, adherens, and tight junction proteins. J Biol Chem 281: 35554 –
35566, 2006. doi:10.1074/jbc.M607305200.
138. Vandenbroucke E, Mehta D, Minshall R, Malik AB. Regulation of
endothelial junctional permeability. Ann N Y Acad Sci 1123: 134 –145,
2008. doi:10.1196/annals.1420.016.
139. Verin AD, Birukova A, Wang P, Liu F, Becker P, Birukov K, Garcia
JG. Microtubule disassembly increases endothelial cell barrier dysfunc-
tion: role of MLC phosphorylation. Am J Physiol Lung Cell Mol Physiol
281: L565–L574, 2001. doi:10.1152/ajplung.2001.281.3.L565.
140. Voelkel NF. Cigarette smoke is an endothelial cell toxin. Am J Respir
Crit Care Med 197: 274, 2018. doi:10.1164/rccm.201706-1123LE.
141. Wacharasint P, Fuengfoo P, Sukcharoen N, Rangsin R; THAI-SICU
Trial Group. Smoking increased risk of ARDS in surgical critically ill
patients: results from the multicenter THAI-SICU study (Abstract). Crit
Care 19, Suppl 1: 238, 2015. doi:10.1186/cc14318.
142. Wagner K, Gröger M, McCook O, Scheuerle A, Asfar P, Stahl B,
Huber-Lang M, Ignatius A, Jung B, Duechs M, Möller P, Georgieff M,
Calzia E, Radermacher P, Wagner F. Blunt chest trauma in mice after
cigarette smoke-exposure: effects of mechanical ventilation with 100% O2.
PLoS One 10: e0132810, 2015. doi:10.1371/journal.pone.0132810.
143. Wagner L, Laczy B, Tamaskó M, Mazák I, Markó L, Molnár GA,
Wagner Z, Mohás M, Cseh J, Fekete A, Wittmann I. Cigarette
smoke-induced alterations in endothelial nitric oxide synthase phosphor-
ylation: role of protein kinase C. Endothelium 14: 245–255, 2007.
doi:10.1080/10623320701606707.
144. Wang B, Rao YH, Inoue M, Hao R, Lai CH, Chen D, McDonald SL,
Choi MC, Wang Q, Shinohara ML, Yao TP. Microtubule acetylation
amplifies p38 kinase signalling and anti-inflammatory IL-10 production.
Nat Commun 5: 3479, 2014. doi:10.1038/ncomms4479.
145. Ward C, Thien F, Secombe J, Gollant S, Walters EH. Bronchoalveo-
lar lavage fluid urea as a measure of pulmonary permeability in healthy
AJP-Lung Cell Mol Physiol • doi:10.1152/ajplung.00373.2017 • www.ajplung.org
Downloaded from journals.physiology.org/journal/ajplung (005.108.050.203) on May 21, 2023.
smokers. Eur Respir J 15: 285–290, 2000. doi:10.1034/j.1399-3003.
2000.15b11.x.
146. Ware LB, Lee JW, Wickersham N, Nguyen J, Matthay MA, Calfee
CS; California Transplant Donor Network. Donor smoking is associ-
ated with pulmonary edema, inflammation and epithelial dysfunction in
ex vivo human donor lungs. Am J Transplant 14: 2295–2302, 2014.
doi:10.1111/ajt.12853.
147. Ware LB, Zhao Z, Koyama T, May AK, Matthay MA, Lurmann FW,
Balmes JR, Calfee CS. Long-term ozone exposure increases the risk of
developing the acute respiratory distress syndrome. Am J Respir Crit Care
Med 193: 1143–1150, 2016. doi:10.1164/rccm.201507-1418OC.
148. Wickenden JA, Clarke MC, Rossi AG, Rahman I, Faux SP, Donald-
son K, MacNee W. Cigarette smoke prevents apoptosis through inhibi-
tion of caspase activation and induces necrosis. Am J Respir Cell Mol
Biol 29: 562–570, 2003. doi:10.1165/rcmb.2002-0235OC.
149. Winkler R, Benz V, Clemenz M, Bloch M, Foryst-Ludwig A, Wardat
S, Witte N, Trappiel M, Namsolleck P, Mai K, Spranger J, Matthias
G, Roloff T, Truee O, Kappert K, Schupp M, Matthias P, Kintscher
U. Histone deacetylase 6 (HDAC6) is an essential modifier of glucocor-
ticoid-induced hepatic gluconeogenesis. Diabetes 61: 513–523, 2012.
doi:10.2337/db11-0313.
150. Wong CM, Yang L, Chan KP, Chan WM, Song L, Lai HK, Thach
TQ, Ho LM, Chan KH, Lam TH, Peiris JS. Cigarette smoking as a risk
factor for influenza-associated mortality: evidence from an elderly co-
hort. Influenza Other Respi Viruses 7: 531–539, 2013. doi:10.1111/j.
1750-2659.2012.00411.x.
150a.World Health Organization. Tobacco Fact Sheet [Online]. Geneva: WHO.
http://www.who.int/mediacentre/factsheets/fs339/en/ [March 2018].
151. Wright JL, Tai H, Churg A. Vasoactive mediators and pulmonary hyper-
tension after cigarette smoke exposure in the guinea pig. J Appl Physiol
(1985) 100: 672–678, 2006. doi:10.1152/japplphysiol.00274.2005.
152. Wright JL, Zhou S, Churg A. Pulmonary hypertension and vascular
oxidative damage in cigarette smoke exposed eNOS⫺/⫺ mice and human
smokers. Inhal Toxicol 24: 732–740, 2012. doi:10.3109/08958378.2012.
715698.
153. Wu MH. Endothelial focal adhesions and barrier function. J Physiol 569:
359 –366, 2005. doi:10.1113/jphysiol.2005.096537.
154. Xiong Y, Zhao K, Wu J, Xu Z, Jin S, Zhang YQ. HDAC6 mutations
rescue human tau-induced microtubule defects in Drosophila. Proc Natl
Acad Sci USA 110: 4604 –4609, 2013. doi:10.1073/pnas.1207586110.
155. Yao H, Rahman I. Role of histone deacetylase 2 in epigenetics and
cellular senescence: implications in lung inflammaging and COPD. Am J
Physiol Lung Cell Mol Physiol 303: L557–L566, 2012. doi:10.1152/
ajplung.00175.2012.
156. Yildiz L, Akçay F, Kaynar H, Bakan N. Increased plasma endothelin-1
in heavy and light smokers. Clin Chem 42: 483–484, 1996.
157. Zhang L, Jin S, Wang C, Jiang R, Wan J. Histone deacetylase
inhibitors attenuate acute lung injury during cecal ligation and puncture-
induced polymicrobial sepsis. World J Surg 34: 1676 –1683, 2010.
doi:10.1007/s00268-010-0493-5.
158. Zhang Y, Kwon S, Yamaguchi T, Cubizolles F, Rousseaux S,
Kneissel M, Cao C, Li N, Cheng HL, Chua K, Lombard D, Mizeracki
A, Matthias G, Alt FW, Khochbin S, Matthias P. Mice lacking
histone deacetylase 6 have hyperacetylated tubulin but are viable and
develop normally. Mol Cell Biol 28: 1688 –1701, 2008. doi:10.1128/
MCB.01154-06.
159. Zhang Y, Li N, Caron C, Matthias G, Hess D, Khochbin S, Matthias
P. HDAC-6 interacts with and deacetylates tubulin and microtubules in
vivo. EMBO J 22: 1168 –1179, 2003. doi:10.1093/emboj/cdg115.
160. Zhao X, Peng X, Sun S, Park AY, Guan JL. Role of kinase-indepen-
dent and -dependent functions of FAK in endothelial cell survival and
barrier function during embryonic development. J Cell Biol 189: 955–
965, 2010. doi:10.1083/jcb.200912094.
161. Zhou Y, Mohsenin A, Morschl E, Young HW, Molina JG, Ma W,
Sun CX, Martinez-Valdez H, Blackburn MR. Enhanced airway in-
flammation and remodeling in adenosine deaminase-deficient mice lack-
ing the A2B adenosine receptor. J Immunol 182: 8037–8046, 2009.
doi:10.4049/jimmunol.0900515.
162. Zhou Y, Murthy JN, Zeng D, Belardinelli L, Blackburn MR. Alter-
ations in adenosine metabolism and signaling in patients with chronic
obstructive pulmonary disease and idiopathic pulmonary fibrosis. PLoS
One 5: e9224, 2010. doi:10.1371/journal.pone.0009224.