 ) and Activated Neutrophils in
Isolated Perfused Rabbit Lungs
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
ABSTRACT |
|---|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
The aim of this study was to examine the effect of ONO-5046, a neutrophil elastase (NE) inhibitor, on
a model of acute lung injury induced by tumor necrosis factor 
(TNF) and phorbol myristate acetate (PMA)-activated neutrophils in isolated perfused rabbit lungs. 120 min after TNF (4,000 JRU/ml)
was injected into the pulmonary artery (PA), 5 × 107 PMA-stimulated neutrophils were infused into
the PA together with 125I-rabbit serum albumin (RSA). In the ONO-5046-treated group (ONO), ONO-5046 (20 mg/kg/h) was continuously infused during the experimental period from 30 min prior to
neutrophil administration. Saline, the ONO-5046 vehicle, was infused instead of ONO-5046 in the
positive control group (ALD) and nonactivated neutrophils were infused without TNF in the negative control group (Cont). PA pressure was monitored over a 240 min period, and bronchoalveolar lavage (BAL) was performed at the end of the experiment. Lung tissues were examined immunohistochemically for the expression of thrombomodulin (TM). The levels of TM in the perfusate were also
measured by ELISA and the radioactivities in the BAL fluid, lung tissue and perfusate were determined to calculate the permeability index (PI) as an indicator of alveolar septal or vascular endothelial damage. The rabbit lungs infused with ONO-5046 showed slower and less increases in PA pressure compared with ALD group. The PI was significantly higher in ALD group (PIBAL = 0.028 ± 0.014, PILUNG = 0.04 ± 0.003) than Cont (PIBAL = 0.002 ± 0.001, PILUNG = 0.015 ± 0.003) and ONO group
(PIBAL = 0.004 ± 0.003, PILUNG = 0.028 ± 0.003 (p < 0.05). ALD group had higher TM levels in the
perfusate and showed decreased expression of TM on the vascular endothelium compared to Cont
and ONO group, suggesting that there was shedding of TM on endothelium and ONO-5046 attenuated a shedding of TM. In conclusion, ONO-5046 attenuated acute lung injury by inhibiting the alveolar epithelial and vascular endothelial injury triggered by activated neutrophils. NE appears to play
an important role in the neutrophil-induced increase of pulmonary epithelial and microvascular permeability observed in acute lung injury.
| |
INTRODUCTION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Acute lung injury in acute respiratory distress syndrome (ARDS) is considered an acute inflammatory condition by many clinicians. Although the pathology of this syndrome is poorly defined, recent evidence suggests that neutrophil elastase (NE) in neutrophils plays a role in the development of acute lung injury, which is characterized by pulmonary edema due to increased permeability (1, 2). NE is thought to play an important role in microvascular endothelial cell damage (3).
Thrombomodulin (TM) is a glycoprotein located on the surface of vascular endothelial cells and functions as an anticoagulant factor by binding thrombin and accelerating thrombin-catalyzed activation of protein C (4, 5). Recent reports have shown that the levels of soluble TM antigen were elevated in plasma of diseases such as diabetes mellitus associated with microangiopathy, disseminated intravascular coagulation, acute respiratory distress syndrome and systemic lupus erythematosus (6). These lines of evidence suggest that the elevation of TM antigen levels in plasma is the reflection of damaged endothelial cells.
We studied epithelial and vascular permability and TM levels in the perfusate of BAL fluid as well as TM expression on
endothelial cells to elucidate the effect of ONO-5046, a novel
NE inhibitor (9), on a model of acute lung injury induced by
TNF and PMA in isolated perfused rabbit lungs.
| |
METHODS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Reagents
ONO-5046(N-[2-[4-(2,2 dimethylpropionyloxy) phenylsulfonylamino]
benzoyl] amino acetic acid) was synthesized and generously supplied
by the chemistry laboratories of Ono Pharmaceutical Co., Ltd. (Osaka, Japan). Recombinant human TNF was kindly supplied from
Dainippon Pharmaceutical Co., Ltd. (Osaka, Japan); its specific activity was 2.55 × 106 Japan reference unit (JRU)/mg protein, JRU was
defined as the producing 50% cytotoxicity of L-M fibroblast cells
(American Type Culture Collection, CCL 1.2, Rockville, MD) (10) and
its endotoxin contents was < 0.04 ng/mg protein by standard Limulus
assay. Phorbol myristate acetate (12-O-tetradecanoyl-phorbol-13-acetate) (PMA), was dissolved in dimethyl sulfoxide at 1 mg/ml), rabbit serum albumin (RSA) and Histopaque 1077 were purchased from Sigma
Chemical Co. (St. Louis, MO). 125I-sodium was purchased from American Radiolabeled Chemicals Inc. (St. Louis, MO) and used to label by
IODO-GENTM (PIERCE, Rockford, IL) reagent method. Free 125I was
removed by extensive dialysis at 4° C against physiological saline just
before use. Mouse monoclonal antibodies (mAbs) to rabbit TM (Li-34 and Li-86); a gift from Daiichi Radioisotope Labs., Ltd. (Chiba, Japan), were used for enzyme-linked immunosorbent assay (ELISA)
(11) and immunohistochemical studies.
Isolated Perfused Rabbit Lung Preparations
Japanese white rabbits (2.5-3.0 kg) were anesthetized with ketamine hydrochloride (20 mg/kg, intramuscular) and sodium pentobarbital (30 mg/kg, intravenously). They were heparinized (1,000 units/kg) through a catheter inserted into a carotid artery and exsanguinated. The chest was then rapidly opened and the pulmonary artery (PA), left atrium, and trachea were cannulated. The lungs were then removed en bloc with the heart and placed in an artificial thorax. The pulmonary circulation from pulmonary artery to vein was washed gently with 50 ml heparinized physiological saline. The arterial and venous cannulas were connected to the perfusion system which had been primed with 80 ml of Krebs' Ringer bicarbonate buffer containing 5% bovine serum albumin (BSA) and glucose. The perfusate was pumped at a constant rate (20 ml/min) into the pulmonary artery from a reservoir by the peristaltic pump (EYELA Roller Pump RP-30; Rikakikai Co., Ltd., Tokyo, Japan). It then drained from the left atrium back into the reservoir. The temperature on the perfusion system was maintained at 37° C in the water bath. The pulmonary artery and the left atrial pressure were monitored throughout the perfusion period using a pressure transducer (RMP-6004; Nihon Kohden, Tokyo, Japan) and the reference level for vascular pressure was the middle of the lungs. The lungs were passively ventilated at a frequency of 18/min by the fluctuating negative pressure that was generated using an animal respirator (Model SN-480-3; Shinano Seisakusho, Tokyo, Japan) and a vacuum pump as shown in Figure 1. The tracheal cannula was connected to a meteorological balloon containing a gas mixture of 16% O2, 6% CO2, and 78% N2, which maintained the perfusate PO2 at about 100 mm Hg, PCO2 at about 40 mm Hg and pH at 7.4.
|
Preparation of Neutrophils
Eighty milliliters of autologous heparinized blood was mixed with 40 ml of 1.5% dextran-phosphate buffered saline (PBS, Ca2+ and Mg2+ free) and divided into four plastic tubes. The tubes were incubated for 45 min at room temperature to sediment red blood cells. The leukocyte-rich supernatant was layered onto 15 ml Histopaque 1077 (Sigma) (15 ml supernatant per 15 ml Histopaque 1077). The mixture was centrifuged at 500 g for 30 min at room temperature. The neutrophil pellet was suspended in 50 ml PBS. The PMA (50 ng/ml) was then added and incubated for 30 min in a water bath at 37° C (12). This PMA concentration is similar to that used by others in isolated perfused lungs (13, 14). This mixture was centrifuged at 1,500 g for 30 min. The supernatant was removed and the pellet was washed with 20 ml PBS followed by centrifugation at 1,500 g for 10 min. After resuspension in 10 ml PBS, actually 5 × 107 neutrophils (> 90% neutrophils, > 99% viable by trypan-blue exclusion, no aggregation) without PMA and platelets were perfused through the PA for 10 min.
The PMA-stimulated Neutrophil-elastase Production of Neutrophils
Neutrophils were prepared as stated above. 1 × 106 neutrophils were
suspended in 10 ml rabbit plasma and added at four different concentrations (0, 25, 50, 100 ng/ml) of PMA, and incubated for 30 min at
37° C. These mixtures were centrifuged at 1,500 g for 30 min. The NE-
1-proteinase inhibitor complex in supernatants were measured by
an ELISA kit, PMN Elastase (Merck, Germany).
Experimental Protocol
ONO group (n = 5). The general experimental protocol is shown in
Figure 2. The lungs were treated with TNF (4,000 JRU/ml) for 120 min by injection into the pulmonary artery after the initial baseline
measurements. 5 × 107 neutrophils, stimulated with PMA (50 ng/ml)
described above, were then infused into the PA for 10 min (15). 50 µCi/ml of 125I-rabbit serum albumin (RSA) was added to the perfusate. ONO-5046 (20 mg/kg/h) was continuously infused through the
PA during the experimental period from 30 min prior to neutrophil
administration. PA pressure was monitored over a 4-h period. At the
end of the experiment, the lungs were perfused with 100 ml physiological saline to wash out 125I-RSA in the vasculature (16), then right and
left lungs were weighted and BAL was performed. For a BAL, 20 ml
of sterile 0.9% sodium chloride were instilled into the left lung through
the left main bronchus and aspirated by gentle syringe suction. This procedure was repeated twice (total volume of 60 ml instilled) (14, 17).
One gram of minced right lung was taken from each animal and the
radioactivity was determined by a gamma counter.
|
Acute lung damage (ALD) group (n = 5). For this group, instead of ONO-5046, normal saline was continuously infused via the PA during the experimental period.
Control group (n = 4). For this group, instead of PMA-stimulated neutrophils, only unstimulated neutrophils were infused into PA for 10 min.
Measurement of Permeability Index
Transvascular flux of 125I-RSA was assessed by the ratio of total lung tissue activities to the activities in the perfusate, which was used to estimate vascular endothelial damage and termed permeability index in lung (PILUNG). Transalveolar flux of 125I-RSA was assessed by the ratio of the activities in the BAL fluid calculated in terms of lung weight to that in the perfusate, which was used to estimate alveolar septal damage and called permeability in BAL (PIBAL) (16, 18, 19).
The ratio of free 125I to bound albumin was less than 1% as 125I- labeled RSA was dialyzed against distilled water just before the experiment. Therefore, the results of permeability index were determined without influences of free 125I.
Measurement of Thrombomodulin (TM)
At the end of the experiment, the levels of TM in the perfusate and BAL were measured by ELISA, using two anti-TM mAbs, Li-34 and Li-84. These two antibodies were purified from mouse IgG anti-rabbit lung TM (American Diagnostica Inc., Co., New York, NY) antibody in the Daiichi Radioisotope Labs., Ltd. They had no complementary and inhibitory interactions with each other. Li-84 was used to coat the plates and Li-34 as horseradish peroxidase (HRP)-conjugated antibody, Li-34 IgG was labeled by the oxidation of periodic acid method. The titration curve of TM, using rabbit lung TM in buffer was in a linear fashion in the range from 0 to 10 ng/ml (data not shown). ELISA method was the same as described previously (6).
Statistics
The data were analyzed using Mann-Whitney U-test and Kruskal-Wallis test, and a p value less than 0.05 was considered statistically significant. Data are expressed as means ± SD.
| |
RESULTS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
NE Production of Neutrophils Stimulated with PMA
There were significant positive correlations between NE release from neutrophils and PMA concentrations in the range from 0 to 100 ng/ml of PMA (r = 0.890, p < 0.0001) (Figure 3). Since the neutrophils were aggregated at concentrations > 100 ng/ml of PMA, we used the 50 ng/ml in the experiments.
|
PA Pressure
Pulmonary artery pressure in three groups is shown in Figure 4. There were no differences in baselines among them. In ALD group, PA pressure increased rapidly from 15 min after neutrophil infusion and reached a peak at 60 min. ONO group showed slower and lesser increases in PA pressure compared with the ALD group at 15, 30, 45, 60 min (p < 0.05).
|
PA pressure did not increase significantly, either in the group
infused only with PMA-stimulated neutrophils or in the group infused with unstimulated neutrophils following TNF (data
not shown).
Permeability Index in BAL
The experiments were terminated 2 h after neutrophil injection and the permeability of BAL and lung parenchyma were measured. The mean negative (Cont group) and positive (ALD group) control permeability indices in BAL were 0.002 ± 0.001 and 0.028 ± 0.014, respectively. The permeability index in BAL of ALD group was significantly higher than Cont group. ONO-5046 treatments (ONO group) were highly protective, reducing the permeability indices (0.004 ± 0.003) by 84% (p < 0.05) (Figure 5).
|
The permeability indices in BAL did not increase either in
the group infused only with PMA-stimulated neutrophils
(PIBAL = 0.002 ± 0.001), or in the group infused with unstimulated neutrophils following TNF, compared with the negative control group (data not shown).
Permeability Index in Lung
The permeability indices in lung of Cont group and ALD group were 0.015 ± 0.003 and 0.04 ± 0.003, respectively. The permeability index in lung of ALD group was significantly higher than Cont group. ONO group moderately reduced the permeability indices (0.028 ± 0.003) by 30% (p < 0.001) (Figure 6).
|
The permeability indices in lung did not increase in either
groups of only PMA-stimulated neutrophils infused (PILUNG = 0.017 ± 0.001), nor TNF unstimulated neutrophils infused
following TNF compared with the negative control group (data
not shown).
The Ratio of TM to Protein Levels in BAL
The TM levels in BAL were corrected by the protein concentrations in BAL. In Cont group and ALD group, the ratio of TM to protein levels in BAL were 2.5 ± 1.9 and 8.2 ± 3.8, respectively. There were no differences between ALD and Cont group in the ratio of TM to protein levels in BAL. ONO group did not reduce the ratio as compared with ALD (p < 0.05) (Figure 7).
|
TM Level in Perfusate
In Cont group and ALD group, TM levels in the perfusate were 135.3 ± 74.1 and 353.1 ± 154.3 ng/ml, respectively. TM levels in ALD group significantly increased than in Cont group, and ONO group showed significantly reduced TM levels (114.1 ± 37.1) by 68% (p < 0.01) (Figure 8).
|
| |
DISCUSSION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
In the present study, we showed that ONO-5046, a novel NE
inhibitor, attenuated acute lung injury induced by TNF and
PMA-activated neutrophils in isolated perfused rabbit lungs.
The release of TNF during septicemia (20) has been linked
to neutrophil-mediated vascular injury (21). We employed a
concentration of 1,600 ng/ml of TNF infused over 120 min,
which is about 1,000-fold in excess of what may be encountered in sera of patients with septicemia (20, 22) and ARDS
(23). However, this concentration of TNF together with non-activated neutrophils did not induce pulmonary vascular endothelial leakage (data not shown). In vitro, TNF enhanced
pulmonary microvascular endothelial cell adhesiveness and
resultant endothelial hyperadhesiveness potentiated the activated neutrophil-mediated increase in endothelial permeability (24). In our experiments, both TNF priming of endothelial cells and PMA-activated neutrophils were necessary to
induce acute lung injury. These results may reflect the higher
levels of TNF in a local environment between alveolar space
to endothelium than in a plasma, because higher levels of
TNF in BAL fluid were observed than in sera of patients
with ARDS (25).
PMA is a potent activator of neutrophils. Shasby and coworkers (14) have shown that neither neutrophils nor PMA
alone caused edema in isolated perfused lungs but that the activation of the neutrophils by addition of PMA resulted in severe acute edema. However, when a high concentration of
PMA (57 ng/ml) devoid of neutrophils was added to the perfusate, perfusion pressure and lung weight increased in isolated
rat lungs (26). Therefore, in order to activate the neutrophils,
we avoided infusing PMA directly into the perfusate, but preincubated neutrophils with PMA (50 ng/ml) (12) and then
only activated neutrophils were infused into the PA. In the
present study, infusion of activated-neutrophils into the PA,
without TNF, did not induce acute lung injury. In vitro, the
release of NE from neutrophils was increased by PMA (Figure 3).
Kawabata and colleagues reported the biochemical and pharmacological properties of ONO-5046. The inhibitory activity
of ONO-5046 was studied using a synthetic substrate, suc-Ala-Pro-Ala-p-nitrosoaniline(pNa). ONO-5046 potently inhibited
human neutrophil elastase (HNE) with an IC50 of 0.044 ± 0.003 µM, Ki of 0.20 ± 0.02 µM (suc-Ala-Pro-Ala pNa as a
substrate). The inhibition is competitive as determined by Lineweaver-Burk plot analysis. ONO-5046 also inhibited leukocyte elastase obtained from rabbits, rats, hamsters or mice
with IC50 values comparable to that against HNE. Furthermore,
ONO-5046 was inactive against bovine pancreatic trypsin, human plasma thrombin, human plasma plasmin, porcine pancreas
kallikrein, human plasma kallikrein, bovine pancreas chymotrypsin and human neutrophil cathepsin G indicating that inhibition by ONO-5046 is highly specific to neutrophil elastase.
Enhanced capillary permeability induced by HNE has been reported by Kawabata. Guinea pigs were injected with HNE in
the clipped-back skin. ONO-5046 was administered intravenously before HNE injection. The increase of skin capillary permeability in guinea pig was directly proportional to the amount
of HNE, in the range of 5 × 10
3 to 40 × 103 U HNE. Intravenous administration of ONO-5046 significantly and dose dependently suppressed the increase of skin capillary permeability induced by HNE (9). In the present study, NE activities in the perfusate were increased from 2.1 to 24.9 µM in vehicle
treated lungs (ALD). However, they were not increased in the
range of 0.1 to 0.3 µM pNa release in ONO-5046-treated lung
during the experiment. We believed that ONO-5046, a specific
neutrophil elastase inhibitor, would attenuate the neutrophil-dependent pulmonary edema in isolated perfused rabbit lungs.
ONO-5046 is a novel inhibitor of human neutrophil elastase,
and it has also the same inhibitory effect on NE of rabbit elastase
(9). Recent studies showed that ONO-5046 inhibited LT-B4 induced lung injury in isolated, perfused rabbit lungs (27), endotoxin-induced lung injury in sheep (28) and in guinea pigs (19).
The fate of the neutrophils was already reported by Sakamaki and coworkers (19) showing that the increased neutrophils in the lung tissue were unchanged, but neutrophil accumulation in the BAL fluid was reduced by ONO-5046 in endotoxin-induced acute lung injury. In our study, neutrophils in the perfusate and BAL fluid were not increased in vehicle treated lung, and slight thickening of alveolar septa and neutrophil infiltration in the alveolar septa in vehicle and ONO-5046 treated lung was observed. These results implied that neutrophil migration into alveolar spaces did not happen, in contrast neutrophil margination to the pulmonary capillary endothelium occurred. Therefore, most of injected neutrophils are likely to accumulate in the lung.
We used the permeability index in lung as a parameter of transvascular albumin leakage, which reflects mainly pulmonary vascular endothelial leakage. As the alveolar septum consists of endothelial and epithelial barriers, permeability in BAL would reflect both endothelial and epithelial damage (19). ONO-5046 treatment attenuated increases in the permeability index in BAL by 84% (Figure 5), whereas there was 30% less reduction in the permeability index in the lungs than the permeability index in BAL (Figure 6). These findings suggest that ONO-5046 influences the effect on neutrophil-dependent epithelial damage with less of an effect on neutrophil- dependent endothelial damage.
ONO-5046 had no effect on the production of oxygen radical from normal human neutrophil in the range of 1 to 100 µg/ ml. Recent reports have shown that oxygen radicals released from PMA-activated neutrophils might have contributed to the development of acute edematous lung injury in isolated rabbit lungs (14) and antioxidants attenuated acute lung injury in isolated perfused rat lungs (32). Therefore, antioxidants are anticipated to be preventive for the development of acute lung injury in our experimental system. However, this possibility is out of the scope of our present study.
TM is an endothelial cell membrane glycoprotein that contributes to the regulation of coagulation system by binding and accelerates the thrombin-catalyzed activation of protein C (4, 5, 29). Recent reports have suggested that the elevation of TM antigen levels in plasma was the reflection of damaged endothelial cells (6). In the present study, TM expression on endothelial cells in ALD group was significantly decreased from the control group (data not shown). On the other hand, TM levels in the perfusate of ALD group was higher than that of control group. ONO-5046 attenuated a decrease of TM expression on endothelial cells and an increase of TM levels in the perfusate in ALD group (Figure 8). These findings suggest that more TM on the endothelium had been shed to the perfusate in ALD group compared with control group and that ONO-5046 reduced the shedding of TM from the endothelium.
The expression of TM is known to be influenced by PMA,
TNF, or a combination of these two agents. They reduce TM
expression on cells by endocytosis and lysosomal degredation
(30, 31). Since no PMA was in the perfusate in our model, endocytosis and degradation of TM by PMA are not responsible
for a decrease of TM expression. TM antigen levels dropped
to 80% of control values within four hours after TNF treatment (31), whereas TM levels in the perfusate in ALD group
significantly increased compared to the Cont and ONO group.
These observations show that a decrease of TM expression on
endothelium was elicited by the shedding of TM to the perfusate as a result of endothelial cell damage, and not by down
regulation with TNF.
In conclusion, the present study suggested that ONO-5046, a potent NE inhibitor, attentuated acute lung injury by inhibiting the alveolar epithelial and vascular endothelial injury triggered by activated neutrophils and reduced the shedding of TM on the endothelium. Neutrophil elastase appears to play an important role in the neutrophil-induced increase of pulmonary microvascular permeability observed in acute lung injury. ONO-5046 has been proven to be safe and a specific neutrophil elastase inhibitor (9). Our results suggest that the neutrophil elastase inhibitor is promising in the treatment of ARDS.
| |
Footnotes |
|---|
Correspondence and requests for reprints should be addressed to Dr. Yasuyuki Yoshizawa, First Department of Internal Medicine, Tokyo Medical and Dental University, 5-45, Yushima 1-chome, Bunkyo-ku, Tokyo 113, Japan. E-mail: yoshi. med1{at}med.tmd.ac.jp
(Received in original form December 2, 1996 and in revised form August 12, 1997).
The authors are deeply grateful to Dr. Vernon L. Moore (Merck Research Laboratories, Rahway, NJ) for critically reviewing the manuscript.Acknowledgments: ONO-5046 was synthesized and generously supplied by the chemistry laboratories of Ono Pharmaceutical Co., Ltd. (Osaka, Japan).
This work was supported by the research committee of the Japanese Ministry of Health and Welfare on interstitial pulmonary disease and by grant-in-aid for scientific research 07670661 from the Japanese Ministry of Education.
| |
References |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
1. Weiss, S. T.. 1989. Tissue destruction by neutrophils. N. Engl. J. Med. 320: 365-376 [Medline].
2. Tate, R. M., and J. E. Repine. 1983. Neutrophils and the adult respiratory distress syndrome. Am. Rev. Respir. Dis. 128: 552-559 [Medline].
3. Smedley, L. A., M. G. Tonnesen, C. H. Sandhaus, and G. S. Worthen. 1986. Neutrophil-mediated injury to endothelial cells: enhancement by endotoxin and essential role of neutrophil elastase. J. Clin. Invest. 77: 1233-1243 .
4.
Esmon, C. T., and
W. G. Owen.
1981.
Identification of an endothelial
cell cofactor for thrombin-catalyzed activation of protein C.
Proc.
Natl. Acad. Sci. U.S.A.
78:
2249-2252
5.
Dittman, W. A., and
P. W. Majerus.
1990.
Structure and function of
thrombomodulin: a natural anticoagulant.
Blood
75:
329-336
6.
Takano, S.,
S. Kimura,
S. Ohdama, and
N. Aoki.
1990.
Plasma thrombomodulin in health and diseases.
Blood
76:
2024-2029
7. Iwashima, Y., T. Sato, K. Watanabe, and I. Marino. 1990. Elevation of plasma thrombomodulin level in diabetic patients with early diabetic nephropathy. Diabetes 39: 983-988 [Abstract].
8. Ohdama, S., S. Takano, S. Miyake, S. Kubota, K. Sato, and N. Aoki. 1994. Plasma thrombomodulin as a marker of vascular injuries in collagen vascular disease. Am. J. Clin. Pathol. 101: 109-113 [Medline].
9. Kawabata, K., M. Suzuki, M. Sugitani, K. Imaki, M. Toda, and K. Miyamoto. 1991. ONO-5046, a novel inhibitor of human neutrophil elastase. Biochem. Biophys. Res. Commun. 177: 814-820 [Medline].
10. Yamazaki, S., E. Onishi, K. Enami, K. Natori, M. Kohase, H. Sakamoto, M. Tanouchi, and H. Hayashi. 1986. Proposal of standardized methods and reference for assaying recombinant human tumor necrosis factor. Japan. J. Med. Sci. Biol. 39: 105-118 [Medline].
11. Uchiyama, H., H. Ohtani, H. Hiraishi, S. Horie, and M. Kazama. 1992. Changes in plasma thrombomodulin antigen in rabbit developing endotoxin-induced disseminated intravascular coagulation and the effect of heparin. Thromb. Res. 65: 593-604 [Medline].
12. Lindahl, L., A. Ljungman, R. Bruhn, R. Hede, and C. Tagesson. 1991. Calcium ionophore-activated neutrophils prestimulated with endotoxin increase pulmonary arterial pressure and vascular leakage in isolated perfused rat lungs: role of platelet-activating factor. Experimental Lung Research 17: 77-89 [Medline].
13. Patterson, C., J. Bernard, J. Lafuze, M. Hull, and R. Rhoades. 1989. The role of activation of neutrophils and microvascular pressure in acute pulmonary edema. Am. Rev. Respir. Dis. 140: 1052-1062 [Medline].
14. Shasby, D. M., K. M. Vanbenthuysen, R. M. Tate, S. S. Shasby, and J. E. Repine. 1982. Granulocytes mediate acute edematous lung injury in rabbits and in isolated rabbit lungs perfused with phorbol myristate acetate: role of oxygen radicals. Am. Rev. Respir. Dis. 123: 443-447 .
15. Lo, S. K., J. Everitt, J. Gu, and A. B. Malik. 1992. Tumor necrosis factor mediates experimental pulmonary edema by ICAM-1 and CD18- dependent mechanisms. J. Clin. Invest. 89: 981-988 .
16. Mulligan, M. S., M. L. Jones, A. A. Vaporciyan, and P. A. Ward. 1993. Protective effects of IL-4 and IL-10 against immune complex-induced lung injury. J. Immunol. 151: 5666-5674 [Abstract].
17.
Tsukimoto, K.,
N. Yoshimura,
M. Ichioka,
N. Tojo, and
J. B. West.
1994.
Protein, cell, and LTB4 concentrations of lung edema fluid produced
by high capillary pressures in rabbit.
J. Appl. Physiol.
76:
321-327
18.
Ismail, G.,
M. L. Morganroth,
R. F. Todd III, and
L. A. Boxer.
1987.
Prevention of pulmonary injury in isolated perfused rat lungs by activated human neutrophils preincubated with anti-Mo1 monoclonal antibody.
Blood
69:
1167-1174
19. Sakamaki, F., A. Ishizaka, T. Urano, K. Sayama, H. Nakamura, T. Terashima, Y. Waki, S. Tasaka, N. Hasegawa, K. Sato, N. Nakagawa, T. Obata, and M. Kanazawa. 1996. Effect of specific neutrophil elastase inhibitor, ONO-5046, on endotoxin-induced acute lung injury. Am. J. Respir. Crit. Care Med. 153: 391-397 [Abstract].
20. Michie, H. R., K. R. Manogue, D. R. Spriggs, R. Revhaug, S. O. O'Dwyer, C. A. Dinarello, A. Cerami, S. M. Wolff, and D. W. Wilmore. 1988. Detection of circulating tumor necrosis factor after endotoxin administration. N. Engl. J. Med. 318: 1481-1486 [Abstract].
21.
Tracey, K. J.,
B. Beutler,
S. F. Lowry,
J. Merryweather,
S. Wolpe,
I. W. Milsark,
R. J. Harri,
T. J. Fahey III,
A. Zentella,
J. D. Albert,
G. T. Shires, and
A. Cerami.
1986.
Shock and tissue injury induced by recombinant human cachectin.
Science
234:
470-474
22. Damas, P., A. Reuter, P. Gysen, J. Demonty, M. Lamy, and P. Franchimont. 1989. Tumor necrosis factor and interleukin-1 serum levels during severe sepsis in humans. Crit. Care Med. 17: 975-978 [Medline].
23. Roten, R., M. Markert, F. Feihl, M. Shaller, M. Tagan, and C. Perret. 1991. Plasma levels of tumor necrosis factor in the adult respiratory distress syndrome. Am. Rev. Respir. Dis. 143: 590-592 [Medline].
24. Gibbs, L. S., L. Lai, and A. B. Malik. 1990. Tumor necrosis factor enhances the neutrophil-dependent increase in endothelial permeability. J. Cell Physiol. 145: 496-500 [Medline].
25. Hyers, T. M., S. M. Tricomi, P. A. Dettenmeier, and A. A. Fowler. 1991. Tumor necrosis factor levels in serum and bronchoalveolar lavage fluid of patients with the adult respiratory distress syndrome. Am. Rev. Respir. Dis. 144: 268-271 [Medline].
26. Carpenter, L. J., K. J. Johnson, R. G. Kunkel, and R. A. Roth. 1987. Phorbol myristate acetate produces injury to isolated rat lungs in the presence and absence of perfused neutrophils. Toxicol. Appl. Pharmacol. 91: 22-32 [Medline].
27.
Yoshimura, K.,
S. Nakagawa,
S. Koyama,
T. Kobayashi, and
T. Homma.
1994.
Roles of neutrophil elastase and superoxide anion in leukotriene B4-induced lung injury in rabbit.
J. Appl. Physiol.
76:
91-96
28.
Kubo, K.,
T. Kobayashi,
T. Hayano,
T. Koizumi,
T. Honda,
M. Sekiguchi, and
A. Sakai.
1994.
Effects of ONO-5046, a specific neutrophil
elastase inhibitor, on endotoxin-induced lung injury in sheep.
J. Appl.
Physiol.
77:
1333-1340
29.
Esmon, C. T..
1989.
The roles of protein C and thrombomodulin in the
regulation of blood coagulation.
J. Biol. Chem.
264:
4743-4746
30.
Dittman, W. A.,
T. K. Kumada,
J. E. Sadler, and
P. W. Majerus.
1988.
The structure and function of mouse thrombomodulin: phorbol myristate acetate stimulates degradation and synthesis of thrombomodulin without affecting mRNA levels in hemangioma cells.
J. Biol. Chem.
263:
15815-15822
31.
Moore, K. L.,
C. T. Esmon, and
N. L. Esmon.
1989.
Tumor necrosis factor leads to the internalization and degradation of thrombomodulin from the surface of bovine aortic endothelial cells in culture.
Blood
73:
159-165
32. McDonald, R. J., E. M. Berger, and J. E. Repine. 1987. Neutrophil- derived oxygen metabolites stimulate thromboxane release, pulmonary artery pressure increases, and weight gains in isolated perfused rat lungs. Am. Rev. Respir. Dis. 135: 957-959 [Medline].
This article has been cited by other articles:
 |
|
 |
|
  D. McClenahan, K. Hellenbrand, D. Atapattu, N. Aulik, D. Carlton, A. Kapur, and C. Czuprynski Effects of Lipopolysaccharide and Mannheimia haemolytica Leukotoxin on Bovine Lung Microvascular Endothelial Cells and Alveolar Epithelial Cells Clin. Vaccine Immunol., February 1, 2008; 15(2): 338 - 347. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Mori, I. Nagahiro, T. Osaragi, K. Kotani, H. Nakanishi, Y. Sano, H. Date, and N. Shimizu Addition of a neutrophil elastase inhibitor to the organ flushing solution decreases lung reperfusion injury in rat lung transplantation Eur. J. Cardiothorac. Surg., November 1, 2007; 32(5): 791 - 795. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-M. Cavaillon and D. Annane Invited review: Compartmentalization of the inflammatory response in sepsis and SIRS Innate Immunity, June 1, 2006; 12(3): 151 - 170. [Abstract] [PDF]
|
|
|
|
 |
|
 Y. Kadoi, H. Hinohara, F. Kunimoto, S. Saito, F. Goto, T. Kosaka, and K. Ieta Pilot Study of the Effects of ONO-5046 in Patients with Acute Respiratory Distress Syndrome Anesth. Analg., September 1, 2004; 99(3): 872 - 877. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. E. Marik Pharmacologic Strategies for the Treatment of Acute Respiratory Distress Syndrome: The Horizon is Getting Closer J Intensive Care Med, November 1, 2002; 17(6): 326 - 328. [PDF]
|
|
|
|
 |
|
 M.-P. Jacob, M. Cazaubon, A. Scemama, D. Prie, F. Blanchet, M.-C. Guillin, and J.-B. Michel Plasma Matrix Metalloproteinase-9 as a Marker of Blood Stasis in Varicose Veins Circulation, July 30, 2002; 106(5): 535 - 538. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Takayama, M. Ishibashi, H. Ishii, T. Kuraki, T. Nishida, and M. Yoshida Effects of neutrophil elastase inhibitor (ONO-5046) on lung injury after intestinal ischemia-reperfusion J Appl Physiol, October 1, 2001; 91(4): 1800 - 1807. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. L. LEE and G. P. DOWNEY Leukocyte Elastase . Physiological Functions and Role in Acute Lung Injury Am. J. Respir. Crit. Care Med., September 1, 2001; 164(5): 896 - 904. [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Mathias, C. G. Tribble, J. F. Dietz, R. P. Nguyen, K. S. Shockey, J. A. Kern, and I. L. Kron Aprotinin improves pulmonary function during reperfusion in an isolated lung model Ann. Thorac. Surg., November 1, 2000; 70(5): 1671 - 1674. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Xu, A. Rahman, R. D. Minshall, C. Tiruppathi, and A. B. Malik {beta}2-Integrin Blockade Driven by E-Selectin Promoter Prevents Neutrophil Sequestration and Lung Injury in Mice Circ. Res., August 4, 2000; 87(3): 254 - 260. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Proc. Am. Thorac. Soc. | Am. J. Respir. Cell Mol. Biol. |