Prenatal inflammation exacerbates hyperoxia-induced functional and structural
changes in adult mice
Markus Velten,1,2Rodney D. Britt Jr.,1Kathryn M. Heyob,1Stephen E. Welty,3Britta Eiberger,4
Trent E. Tipple,1and Lynette K. Rogers1
1Center for Perinatal Research, The Research Institute at Nationwide Children’s Hospital, Columbus, Ohio;2Department
of Anesthesiology and Intensive Care Medicine, Rheinische Friedrich-Wilhlems-University, University Medical Center, Bonn,
Germany;3Texas Children’s Hospital, Baylor College of Medicine, Houston, Texas; and4Institute of Anatomy, Rheinische
Friedrich-Wilhlems-University, Bonn, Germany
Submitted 20 January 2012; accepted in final form 9 June 2012
Velten M, Britt Jr. RD, Heyob KM, Welty SE, Eiberger B,
Tipple TE, Rogers LK. Prenatal inflammation exacerbates hyperoxia
induced functional and structural changes in adult mice. Am J Physiol
Regul Integr Comp Physiol 303: R279–R290, 2012. First published
June 20, 2012; doi:10.1152/ajpregu.00029.2012.—Maternally derived
inflammatory mediators, such as IL-6 and IL-8, contribute to preterm
delivery, low birth weight, and respiratory insufficiency, which are
routinely treated with oxygen. Premature infants are at risk for
developing adult-onset cardiac, metabolic, and pulmonary diseases.
Long-term pulmonary consequences of perinatal inflammation are
unclear. We tested the hypothesis that a hostile perinatal environment
induces profibrotic pathways resulting in pulmonary fibrosis, includ-
ing persistently altered lung structure and function. Pregnant C3H/
HeN mice injected with LPS or saline on embryonic day 16. Offspring
were placed in room air (RA) or 85% O2for 14 days and then returned
to RA. Pulmonary function tests, microCTs, molecular and histological
analyses were performed between embryonic day 18 and 8 wk. Alveo-
staining and protein levels were increased, and static compliance was
decreased only in LPS/O2-exposed mice. Three-dimensional microCT
reconstruction and quantification revealed increased tissue densities only
in LPS/O2 mice. Diffuse interstitial fibrosis was associated with de-
creased micro-RNA-29, increased transforming growth factor-? ex-
pression, and phosphorylation of Smad2 during embryonic or early
fetal lung development. Systemic maternal LPS administration in
combination with neonatal hyperoxic exposure induces activation of
profibrotic pathways, impaired alveolarization, and diminished lung
function that are associated with prenatal and postnatal suppression of
pulmonary fibrosis; transforming growth factor-?; fetal origins; mi-
crocomputed tomography scans; pulmonary function tests
THE CAUSES AND CONSEQUENCES of preterm birth remain poorly
understood and present a significant health burden. In the past
three decades, advances in neonatal care, including use of
antenatal corticosteroids, surfactant therapy, and high-fre-
quency ventilation, have significantly improved survival rates
of extremely preterm (?28 wk gestation) and low-birthweight
infants (45). However, little is known about the long-term
physiological consequences of a hostile perinatal environment.
Data currently being collected indicate that preterm infants
surviving to adulthood are at greater risk for the development
of chronic health problems (9–11). Low birth weight, early
gestational age, and respiratory support are highly associated
with interrupted alveolarization and respiratory insufficiency.
These pathologies can progress into the development of bron-
chopulmonary dysplasia (BPD) (29), a disease pathologically
characterized by impaired alveolarization and diffuse intersti-
tial fibrosis (2, 26). Furthermore, whether they develop BPD or
not, extremely immature infants are at increased risk for
developing adult pulmonary pathologies, including emphy-
sema, chronic obstructive pulmonary disease, asthma, or pul-
monary fibrosis (2, 14, 15, 26, 36, 57).
Maternal infections and/or inflammation and the subsequent
inflammatory responses that contribute to preterm delivery can
significantly impact fetal development (16, 18, 19, 22, 58).
Research supporting the “fetal origins of adult disease” hy-
pothesis has focused on cardiovascular and metabolic diseases
(3, 4, 32, 43); however, disordered fetal development has
profound effects on other organs, including the lung. Recently,
Shi and colleagues (47, 48, 56) demonstrated that early expo-
sures during periods of developmental plasticity contribute to
the development of adult pulmonary diseases.
While the mechanisms responsible for the development of
pulmonary pathologies are multifactorial, common diseases are
often characterized by diminished lung function and interstitial
fibrosis. Mechanistically, lung fibrosis is associated with dys-
regulated transforming growth factor-? (TGF-?) expression
and Smad signaling in human patients and bleomycin-treated
mice (6, 7). Previous studies have implicated a crucial temporal
window for TGF-? signaling during lung development, that, if
interrupted, leads to impaired alveolarization and pulmonary
fibrosis (47, 49). TGF-? modulates the expression of profi-
brotic genes through suppression of micro-RNA (miR)-29 that,
in turn, causes increases in TGF-? expression, in a feed-
forward manner (12, 38). miR-29 has been demonstrated to
target proteins regulated by TGF-? and Smad signaling, such
as collagen and matrix-remodeling proteins (12). miRs regulate
the expression of multiple genes by enhancement, suppression,
or destabilization of target RNAs and are increasingly recog-
nized as important contributors to developmental processes and
disease pathogenesis. Furthermore, dysregulation of miRs has
been linked to fibrosis in multiple organs, including the heart
(52), kidney (8), and lung (12). The contributions of maternal
influences on early disruption of fetal TGF-? pathways or miR
expressions, in developing lungs are unknown.
Many animal models that include fetal inflammation or
postnatal hypo- or hyperoxic exposures have been developed to
study newborn lung diseases. Hyperoxic exposure induces
inflammation and disrupts cell proliferation, leading to alveolar
Address for reprint requests and other correspondence: M. Velten, Dept. of
Anesthesiology and Intensive Care Medicine, Rheinische Friedrich-Wilhelms-
Univ., Univ. Medical Ctr., Sigmund-Freud-Str. 25, 53105 Bonn, Germany
Am J Physiol Regul Integr Comp Physiol 303: R279–R290, 2012.
First published June 20, 2012; doi:10.1152/ajpregu.00029.2012.
0363-6119/12 Copyright © 2012 the American Physiological Societyhttp://www.ajpregu.orgR279
dysplasia in newborn rodents (27, 60). However, the long-term
pulmonary consequences of fetal exposures have not been
extensively investigated (30). We have previously reported
developmental alterations in alveolarization and pulmonary
function 14 days after systemic maternal inflammation and
neonatal hyperoxic exposure (53). This model was designed to
mimic the hostile perinatal inflammatory environment often
encountered by prematurely born human infants. In the current
studies, we tested the hypothesis that the combination of
LPS-induced systemic maternal inflammation and postnatal
hyperoxic exposure would result in 1) persistently altered
alveolarization, 2) lung fibrosis, and 3) impaired pulmonary
function in adulthood.
MATERIALS AND METHODS
Animals and exposure. Animal study protocols were approved by
the Institutional Animal Care and Use Committee at The Research
Institute at Nationwide Children’s Hospital, Columbus, OH. All
animals were handled in accordance with National Institutes of Health
guidelines and housed in a “specified pathogen-free” facility. Mice
were housed in our facility at least 7 days before breeding was started,
and pregnancy was time dated by the presence of a vaginal plug.
Pregnant C3H/HeN mice were injected on embryonic day 16 (E16)
with LPS (80 ?g/kg ip, serotype 0111:B4, no. 437627; Calbiochem,
Gibbstown, NJ) or an equal volume of saline. The amount of LPS was
chosen on the basis of preliminary studies to determine the highest
dose that resulted in viable litter of equal size. Each litter of newborn
mice was paired with a litter born to a dam receiving the same E16
treatment, and the pups were pooled and redistributed randomly, as
previously described (53). One of the paired group of pups was
exposed to 85% O2 for 2 wk (saline/O2, LPS/O2) and subsequently
returned to room air (RA), while the corresponding group was
maintained in RA (saline/RA, LPS/RA). Nursing dams were rotated
between their RA and O2litter every 24 h to prevent oxygen toxicity.
Twenty-four hours of RA or oxygen exposure was designated as day
1. The mice were killed at E18, 7 or 14 days, or 8 wk of life, and only
one pup per litter was used at each time point for analyses. One pup
per litter was analyzed per experiment, and equal numbers of males
and females were measured. For the pulmonary function tests, one
male and one female were analyzed from each litter.
Histology. The left lung was inflation fixed with 10% buffered
formalin at a pressure of 25 cm H2O for 15 min. Following paraffin
embedding, the tissue sections were cut, and 4-?m slides were stained
with hematoxylin and eosin (H&E) for morphometric measurements,
Mason’s trichrome, and Picrosirius red (PSR) stain to assess collagen
Immunohistochemistry. Inflation-fixed left lung tissue sections
were cut and 4-?m slides stained for macrophages with Mac3 mono-
clonal antibody (catalog no. 550292, BD Pharmingen, San Diego,
CA) as the primary antibody and rabbit anti-rat (catalog no. BA-4001;
Vector, Burlingame, CA) as a secondary antibody. Macrophage counts
were performed on five nonoverlapping fields per mouse lung tissues and
n ? 5 animals per group using digital image analysis software with
settings for color and size identification (Image Pro Plus 4.0; Media
Cybernetics, Silver Spring, MD).
Western immunoblotting. Proteins were separated on SDS-PAGE
gels and transferred to PVDF membranes. Membranes were probed
with antibodies to collagen I (ab292; Abcam, Cambridge, MA),
collagen III (EMD Millipore, 234189; Millipore, Billerica, MA),
p-Smad2 (no. 3108, Cell Signaling, Danvers, MA), and total Smad2/3
(no. 3102; Cell Signaling). Blots were developed using enhanced
chemiluminescence (ECL Western blotting detection, GE Healthcare,
Chalfont, Buckinghamshire, UK), and expression levels were quanti-
fied using ImageQuant software, version 5.0 (Molecular Dynamics,
Sunnydale, CA). The density of the band for the protein of interest
was normalized to the density of ?-actin protein (no. ab6276; Abcam).
Pulmonary function tests. A SCIREQ FlexiVent (FlexiVent, SCIREQ,
Montreal, Canada) ventilator was used to perform pulmonary function
analyses. Mice were anesthetized with ketamine (200 mg/kg ip) and
xylazine (20 mg/kg), tracheotomized with a 20-gauge cannula (BD
Intramedic, no. 427564; Becton Dickinson, Franklin Lakes, NJ), and
connected to the FlexiVent ventilator. The plane of anesthesia was
sufficient to prevent spontaneous breathing. The mice were ventilated
with a tidal volume of 10 ml/kg at a frequency of 350 breaths/min and
positive end-expiratory pressure of 2 cm H2O to achieve lung volume
similar to spontaneous breathing. Forced oscillation (0.5–19.6 Hz)
was applied for 8 s. Subsequently, dynamic pressure-volume maneu-
vers were performed stepwise, increasing airway pressure to 30 cm
H2O and then reversing the process. For each parameter, three
measurements were assessed and averaged. Measurements were ex-
cluded from analyses if they were disrupted by a spontaneous breath,
and a coefficient of determination of 0.98 was used as the lower limit
for each measurement.
MicroCT imaging. A General Electric Healthcare Xplore Locus
microCT (General Electric, London, Canada) was used for pulmonary
imaging. Mice were anesthetized with ketamine (20 mg/kg ip) and
xylazine (2 mg/kg ip). Respiratory gated microCT images were
acquired at inspiration, as previously described (17). Images were
reconstructed with a nominal isotropic voxel spacing of 90 ?m. Lung
density data obtained from microCTs were normalized to density
standards and converted to Hounsfield units (HU). Five representative
regions of interest were measured, and the average was considered as
the lung density of the individual mouse (40). After performing
microCT in vivo, mice were killed by ketamine (150 mg/kg)–xylazine
(15 mg/kg) overdose; then they were tracheotomized, lungs were air
inflated, and post-mortem high-resolution microCT were performed at
a constant inflation of 30 cm H2O. Three-dimensional images were
reconstructed with a nominal isotropic voxel spacing of 20 ?m and
evaluated by a radiologist blinded to group assignment.
Quantitative real-time PCR. Total RNA was isolated from frozen
lung tissue using an RNeasy mini kit (Qiagen, Valencia, CA). cDNA
was synthesized using a Maxima first-strand cDNA synthesis kit for
RT-qPCR (no. K1641; Thermo Scientific Fermentas, Glen Burnie,
MD). Quantitative real-time PCR was performed using Maxima
SYBR Green/ROX qPCR Master Mix (K0222; Thermo Scientific
Fermentas) and the Mastercycler epgradient Realplex real-time PCR
detection system (Eppendorf, Hamburg, Germany).
ELISA. Frozen lungs were homogenized, and protein concentra-
tions were determined by Bradford assay. TNF-?, IL-6, keratino-
cyte-derived chemokine (KC), and monocyte chemoattractant pro-
tein-1 (MCP-1) levels were measured using ELISA (Duoset ELISA
kits; R&D Systems, Minneapolis, MN), according to the manufac-
turer’s protocols. Absorbances were determined spectrophometri-
cally using a Spectramax M2 Plate Reader (Molecular Devices,
MicroRNA analyses. miR fractions were isolated and enriched from
lungs with Qiagen RNeasy Mini Kits (Qiagen, Valencia, CA). Sub-
sequently, 100-ng enriched miR was reverse-transcribed with SA
Biosciences miR first-strand kit (SA Biosciences, Frederick, MD).
Real-time PCR was performed on an Mastercycler epgradient Real-
plex real-time PCR detection system (Eppendorf, Hamburg, Ger-
many) using RT2SYBR Green qPCR Mastermix with ROX and RT2
miRNA qPCR assay primer sets (SA Biosciences).
Statistics. Analyses were performed using GraphPad PRISM 5
(GraphPad, La Jolla, CA). Data are expressed as means ? SE.
Statistical analyses were performed using a two-way ANOVA with
Bonferroni post hoc or Student’s t-test. P ? 0.05 was considered
FETAL ORIGINS OF ADULT PULMONARY DISEASE
AJP-Regul Integr Comp Physiol • doi:10.1152/ajpregu.00029.2012 • www.ajpregu.org
development and maintenance of alveolar structure (1). Devel-
opmental overexpression of TGF-? results in impaired alveo-
larization and Smad-dependent interstitial fibrosis in monkeys
(51) and rats (20, 21). Increases in IL-1?, TGF-?, and collagen
I mRNA levels were persistent after birth in hyperoxia-exposed
mice. There were additional increases in collagen III in hyper-
oxia-exposed mice, particularly at 7 days. By day 14, IL-1?
levels were similar to controls but TGF-? and collagen I and III
mRNA expression remained elevated solely in the LPS/O2-
exposed mice (Figs. 6 and 7). Additionally, elevated levels in
TGF-? and collagen I and III mRNA were associated with
increases in p-Smad2 protein levels in only the LPS/O2 ex-
posed at day 14.
Previous investigations have linked macrophage-induced
increased IL-1? levels with altered TGF-? and Smad expres-
sions, and subsequent collagen deposition and fibrosis, sug-
gesting that this phenotype more closely resembles impaired
tissue repair rather than acute injury (6, 37). One explanation of
our current findings could be that the LPS exposure in utero
initiates a lung injury that is either unable to resolve or
interrupted during the healing process by the inflammatory
responses to hyperoxia exposure. Regardless, the sustained
increases in MCP-1, IL-6, TNF-?, IL-1?, TGF-?, and collagen
I and III, distinguish the LPS/O2 from the other treatment
groups and are, in part, responsible for the physiological
deficits in lung structure and function observed at 8 wk.
Our data provide the first evidence that TGF-? mRNA levels
in fetal lungs are increased by LPS-induced maternal inflam-
mation and that subsequent hyperoxia exposure caused the
induction to persist. TGF-? regulation of downstream profi-
brotic genes has been linked to suppression of miR-29b ex-
pression (12). Our data indicate that miR-29b is decreased at
E18 in fetal lung tissues from LPS-treated dams (Fig. 7) and is
inversely correlated with TGF-? and collagen I and III expres-
sions. On day 14, miR-29b expression was further suppressed
in LPS/O2mice; however, miR-29b expression was elevated in
saline/O2compared with saline/RA mice probably as a com-
pensatory response. miR-29b reduces expression of proteins
that are regulated by TGF-? stimulation, such as collagen I and
III (12). We hypothesize that elevated mRNA levels of colla-
gen I and III in LPS/O2 on E18 and 14 days are due to
suppression of miR-29b levels. Whether directly or indirectly
through miR-29b, our data suggest that TGF-? expression is
fundamentally involved in the fibrotic phenotype observed in
the LPS/O2-exposed mice.
In summary, the present study demonstrates that systemic
maternal LPS administration induces a profibrotic response in
fetal lung tissue that begins prior to birth and that results in an
ongoing activation of profibrotic pathways when combined
with neonatal hyperoxic exposure. These dual inflammatory
exposures lead to the development of diffuse interstitial fibro-
sis, impaired alveolarization, and compromised pulmonary
mechanics in both adult male and female offspring with no
differences between sexes. Our data suggest that these changes
are mediated through reduction in miR-29b expression, result-
ing in activation of profibrotic pathways, including, but not
exclusive to, TGF-?-mediated signaling. The finding that peri-
natal exposures result in persistently impaired pulmonary phys-
iology in adulthood provides a novel model to investigate the
influence of the perinatal environment on the development of
adult pulmonary diseases.
The authors would like to acknowledge and thank Dr. Loren Wold for his
editorial assistance and Dr. Frederick Long for radiologic interpretations of the
This work was supported by the Deutsche Forschungsgemeinschaft (VE
614/1-1) and the National Institutes of Health (RDB F31HL097619, TET
1K08HL093365-01A2 and LKR R01AT006880).
No conflicts of interest, financial or otherwise, are declared by the authors.
Author contributions: M.V., S.E.W., T.E.T., and L.K.R. conception and
design of research; M.V., R.D.B.J., K.M.H., and B.E. performed experiments;
M.V., R.D.B.J., K.M.H., and L.K.R. analyzed data; M.V., R.D.B.J., T.E.T.,
and L.K.R. interpreted results of experiments; M.V. and L.K.R. prepared
figures; M.V. and L.K.R. drafted manuscript; M.V., R.D.B.J., B.E., T.E.T., and
L.K.R. edited and revised manuscript; M.V., R.D.B.J., S.E.W., B.E., T.E.T.,
and L.K.R. approved final version of manuscript.
1. Alejandre-Alcazar MA, Kwapiszewska G, Reiss I, Amarie OV, Marsh
LM, Sevilla-Perez J, Wygrecka M, Eul B, Kobrich S, Hesse M,
Schermuly RT, Seeger W, Eickelberg O, Morty RE. Hyperoxia mod-
ulates TGF-beta/BMP signaling in a mouse model of bronchopulmonary
dysplasia. Am J Physiol Lung Cell Mol Physiol 292: L537–L549, 2007.
2. Aly H. Respiratory disorders in the newborn: identification and diagnosis.
Pediatr Rev 25: 201–208, 2004.
3. Barker DJ. The developmental origins of adult disease. J Am Coll Nutr
23: 588S–595S, 2004.
4. Barker DJ. The fetal origins of coronary heart disease. Eur Heart J 18:
5. Bastek JA, Gomez LM, Elovitz MA. The role of inflammation and
infection in preterm birth. Clin Perinatol 38: 385–406, 2011.
6. Bonniaud P, Margetts PJ, Ask K, Flanders K, Gauldie J, Kolb M.
TGF-? and Smad3 signaling link inflammation to chronic fibrogenesis. J
Immunol 175: 5390–5395, 2005.
7. Chen H, Zhuang F, Liu YH, Xu B, Del Moral P, Deng W, Chai Y,
Kolb M, Gauldie J, Warburton D, Moses HL, Shi W. TGF-? receptor
II in epithelia versus mesenchyme plays distinct roles in the developing
lung. Eur Respir J 32: 285–295, 2008.
8. Chung AC, Huang XR, Meng X, Lan HY. miR-192 mediates TGF-?/
Smad3-driven renal fibrosis. J Am Soc Nephrol 21: 1317–1325, 2010.
9. Crump C, Sundquist K, Sundquist J, Winkleby MA. Gestational age at
birth and mortality in young adulthood. JAMA 306: 1233–1240, 2011.
10. Crump C, Winkleby MA, Sundquist J, Sundquist K. Risk of asthma in
young adults who were born preterm: a Swedish national cohort study.
Pediatrics 127: e913–e920, 2011.
11. Crump C, Winkleby MA, Sundquist K, Sundquist J. Risk of diabetes
among young adults born preterm in Sweden. Diabetes Care 34: 1109–
12. Cushing L, Kuang PP, Qian J, Shao F, Wu J, Little F, Thannickal VJ,
Cardoso WV, Lu J. miR-29 is a major regulator of genes associated with
pulmonary fibrosis. Am J Respir Cell Mol Biol 45: 287–294, 2011.
13. Dackor RT, Cheng J, Voltz JW, Card JW, Ferguson CD, Garrett RC,
Bradbury JA, Degraff LM, Lih FB, Tomer KB, Flake GP, Travlos GS,
Ramsey RW Jr, Edin ML, Morgan DL, Zeldin DC. Prostaglandin E2
protects murine lungs from bleomycin-induced pulmonary fibrosis and
lung dysfunction. Am J Physiol Lung Cell Mol Physiol 301: L645–L655,
14. Doyle LW, Anderson PJ. Adult outcome of extremely preterm infants.
Pediatrics 126: 342–351, 2010.
15. Fawke J, Lum S, Kirkby J, Hennessy E, Marlow N, Rowell V, Thomas
S, Stocks J. Lung function and respiratory symptoms at 11 years in
children born extremely preterm: the EPICure study. Am J Respir Crit
Care Med 182: 237–245, 2010.
16. Fernandez-Twinn DS, Ozanne SE. Early life nutrition and metabolic
programming. Ann NY Acad Sci 1212: 78–96, 2010.
FETAL ORIGINS OF ADULT PULMONARY DISEASE
AJP-Regul Integr Comp Physiol • doi:10.1152/ajpregu.00029.2012 • www.ajpregu.org
17. Ford NL, Nikolov HN, Norley CJ, Thornton MM, Foster PJ, Dran-
gova M, Holdsworth DW. Prospective respiratory-gated micro-CT of
free breathing rodents. Med Phys 32: 2888–2898, 2005.
18. Frankel S, Elwood P, Sweetnam P, Yarnell J, Smith GD. Birthweight,
body-mass index in middle age, and incident coronary heart disease.
Lancet 348: 1478–1480, 1996.
19. Freak-Poli R, Chan A, Tucker G, Street J. Previous abortion and risk of
pre-term birth: a population study. J Matern Fetal Neonatal Med 22: 1–7,
20. Gauldie J, Galt T, Bonniaud P, Robbins C, Kelly M, Warburton D.
Transfer of the active form of transforming growth factor-?1 gene to
newborn rat lung induces changes consistent with bronchopulmonary
dysplasia. Am J Pathol 163: 2575–2584, 2003.
21. Gauldie J, Kolb M, Ask K, Martin G, Bonniaud P, Warburton D.
Smad3 signaling involved in pulmonary fibrosis and emphysema. Proc Am
Thorac Soc 3: 696–702, 2006.
22. Getahun D, Ananth CV, Oyelese Y, Peltier MR, Smulian JC, Vintzi-
leos AM. Acute and chronic respiratory diseases in pregnancy: associa-
tions with spontaneous premature rupture of membranes. J Matern Fetal
Neonatal Med 20: 669–675, 2007.
23. Goldenberg RL, Culhane JF, Iams JD, Romero R. Epidemiology and
causes of preterm birth. Lancet 371: 75–84, 2008.
24. Guerrero T, Castillo R, Noyola-Martinez J, Torres M, Zhou X,
Guerra R, Cody D, Komaki R, Travis E. Reduction of pulmonary
compliance found with high-resolution computed tomography in irradi-
ated mice. Int J Radiat Oncol Biol Phys 67: 879–887, 2007.
25. Hampton T. Fetal environment may have profound long-term conse-
quences for health. JAMA 292: 1285–1286, 2004.
26. Husain AN, Siddiqui NH, Stocker JT. Pathology of arrested acinar
development in postsurfactant bronchopulmonary dysplasia. Hum Pathol
29: 710–717, 1998.
27. James ML, Ross AC, Bulger A, Philips JB 3rd, Ambalavanan N.
Vitamin A and retinoic acid act synergistically to increase lung retinyl
esters during normoxia and reduce hyperoxic lung injury in newborn mice.
Pediatr Res 67: 591–597, 2010.
28. Jobe AH. Antenatal associations with lung maturation and infection. J
Perinatol 25 Suppl 2: S31–S35, 2005.
29. Jobe AH. The new bronchopulmonary dysplasia. Curr Opin Pediatr 23:
30. Jobe AH, Ikegami M. Antenatal infection/inflammation and postnatal
lung maturation and injury. Respir Res 2: 27–32, 2001.
31. Kaarteenaho-Wiik R, Paakko P, Herva R, Risteli J, Soini Y. Type I
and III collagen protein precursors and mRNA in the developing human
lung. J Pathol 203: 567–574, 2004.
32. Khan OA, Chau R, Bertram C, Hanson MA, Ohri SK. Fetal origins of
coronary heart disease-implications for cardiothoracic surgery? Eur J
Cardiothorac Surg 27: 1036–1042, 2005.
33. Kim DH, Choi CW, Kim EK, Kim HS, Kim BI, Choi JH, Lee MJ,
Yang EG. Association of increased pulmonary interleukin-6 with the
priming effect of intra-amniotic lipopolysaccharide on hyperoxic lung
injury in a rat model of bronchopulmonary dysplasia. Neonatology 98:
34. Kohmura Y, Kirikae T, Kirikae F, Nakano M, Sato I. Lipopolysac-
charide (LPS)-induced intra-uterine fetal death (IUFD) in mice is princi-
pally due to maternal cause but not fetal sensitivity to LPS. Microbiol
Immunol 44: 897–904, 2000.
35. Kumarasamy A, Schmitt I, Nave AH, Reiss I, van der Horst I, Dony
E, Roberts JD Jr, de Krijger RR, Tibboel D, Seeger W, Schermuly
RT, Eickelberg O, Morty RE. Lysyl oxidase activity is dysregulated
during impaired alveolarization of mouse and human lungs. Am J Respir
Crit Care Med 180: 1239–1252, 2009.
36. Kwinta P, Pietrzyk JJ. Preterm birth and respiratory disease in later life.
Expert Rev Respir Med 4: 593–604, 2010.
37. Ma F, Li Y, Jia L, Han Y, Cheng J, Li H, Qi Y, Du J. Macrophage-
stimulated cardiac fibroblast production of IL-6 is essential for TGF?/
Smad activation and cardiac fibrosis induced by angiotensin II. PLos One
7: e35144, 2012.
38. Mott JL, Kurita S, Cazanave SC, Bronk SF, Werneburg NW, Fer-
nandez-Zapico ME. Transcriptional suppression of mir-29b-1/mir-29a
promoter by c-Myc, hedgehog, and NF-?B. J Cell Biochem 110: 1155–
39. Parr DG, Dirksen A, Piitulainen E, Deng C, Wencker M, Stockley RA.
Exploring the optimum approach to the use of CT densitometry in a
randomised placebo-controlled study of augmentation therapy in alpha
1-antitrypsin deficiency. Respir Res 10: 75, 2009.
40. Plathow C, Li M, Gong P, Zieher H, Kiessling F, Peschke P, Kauczor
HU, Abdollahi A, Huber PE. Computed tomography monitoring of
radiation-induced lung fibrosis in mice. Invest Radiol 39: 600–609, 2004.
41. Razzaque MS, Nazneen A, Taguchi T. Immunolocalization of collagen
and collagen-binding heat shock protein 47 in fibrotic lung diseases. Mod
Pathol 11: 1183–1188, 1998.
42. Redente EF, Jacobsen KM, Solomon JJ, Lara AR, Faubel S, Keith
RC, Henson PM, Downey GP, Riches DW. Age and sex dimorphisms
contribute to the severity of bleomycin-induced lung injury and fibrosis.
Am J Physiol Lung Cell Mol Physiol 301: L510–L518, 2011.
43. Rogers LK, Velten M. Maternal inflammation, growth retardation, and
preterm birth: Insights into adult cardiovascular disease. Life Sci 89:
44. Romero R, Espinoza J, Goncalves LF, Kusanovic JP, Friel L, Hassan
S. The role of inflammation and infection in preterm birth. Semin Reprod
Med 25: 21–39, 2007.
45. Saigal S, Doyle LW. An overview of mortality and sequelae of preterm
birth from infancy to adulthood. Lancet 371: 261–269, 2008.
46. Salminen A, Paananen R, Vuolteenaho R, Metsola J, Ojaniemi M,
Autio-Harmainen H, Hallman M. Maternal endotoxin-induced preterm
birth in mice: fetal responses in Toll-like receptors, collectins, and cyto-
kines. Pediatr Res 63: 280–286, 2008.
47. Shi W, Bellusci S, Warburton D. Lung development and adult lung
diseases. Chest 132: 651–656, 2007.
48. Shi W, Warburton D. Is COPD in adulthood really so far removed from
early development? Eur Respir J 35: 12–13, 2010.
49. Shi W, Xu J, Warburton D. Development, repair and fibrosis: what is
common and why it matters. Respirology 14: 656–665, 2009.
50. Sverzellati N, Zompatori M, De Luca G, Chetta A, Bna C, Ormitti F,
Cobelli R. Evaluation of quantitative CT indexes in idiopathic interstitial
pneumonitis using a low-dose technique. Eur J Radiol 56: 370–375, 2005.
51. Tarantal AF, Chen H, Shi TT, Lu CH, Fang AB, Buckley S, Kolb M,
Gauldie J, Warburton D, Shi W. Overexpression of transforming growth
factor-?1 in fetal monkey lung results in prenatal pulmonary fibrosis. Eur
Respir J 36: 907–914, 2010.
52. Thum T, Gross C, Fiedler J, Fischer T, Kissler S, Bussen M, Galuppo
P, Just S, Rottbauer W, Frantz S, Castoldi M, Soutschek J, Kotelian-
sky V, Rosenwald A, Basson MA, Licht JD, Pena JT, Rouhanifard SH,
Muckenthaler MU, Tuschl T, Martin GR, Bauersachs J, Engelhardt
S. MicroRNA-21 contributes to myocardial disease by stimulating MAP
kinase signalling in fibroblasts. Nature 456: 980–984, 2008.
53. Velten M, Heyob KM, Rogers LK, Welty SE. Deficits in lung alveo-
larization and function after systemic maternal inflammation and neonatal
hyperoxia exposure. J Appl Physiol 108: 1347–1356, 2010.
54. Velten M, Hutchinson KR, Gorr MW, Wold LE, Lucchesi PA, Rogers
LK. Systemic maternal inflammation and neonatal hyperoxia induces
remodeling and left ventricular dysfunction in mice. PLos One 6: e24544,
55. Viscardi RM. Perinatal inflammation and lung injury. Semin Fetal Neo-
natal Med 17: 30–35, 2012.
56. Warburton D, Gauldie J, Bellusci S, Shi W. Lung development and
susceptibility to chronic obstructive pulmonary disease. Proc Am Thorac
Soc 3: 668–672, 2006.
57. Wong PM, Lees AN, Louw J, Lee FY, French N, Gain K, Murray CP,
Wilson A, Chambers DC. Emphysema in young adult survivors of
moderate-to-severe bronchopulmonary dysplasia. Eur Respir J 32: 321–
58. Xiong X, Buekens P, Fraser WD, Beck J, Offenbacher S. Periodontal
disease and adverse pregnancy outcomes: a systematic review. BJOG 113:
59. Yamauchi K, Kasuya Y, Kuroda F, Tanaka K, Tsuyusaki J, Ishizaki
S, Matsunaga H, Iwamura C, Nakayama T, Tatsumi K. Attenuation of
lung inflammation and fibrosis in CD69-deficient mice after intratracheal
bleomycin. Respir Res 12: 131, 2011.
60. Yee M, Chess PR, McGrath-Morrow SA, Wang Z, Gelein R, Zhou R,
Dean DA, Notter RH, O’Reilly MA. Neonatal oxygen adversely affects
lung function in adult mice without altering surfactant composition or
activity. Am J Physiol Lung Cell Mol Physiol 297: L641–L649, 2009.
FETAL ORIGINS OF ADULT PULMONARY DISEASE
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