scholarly journals Expression of hydrophilic surfactant proteins by mesentery cells in rat and man

1997 ◽  
Vol 328 (1) ◽  
pp. 251-256 ◽  
Author(s):  
Bernadette CHAILLEY-HEU ◽  
Sandrine RUBIO ◽  
Jean-Philippe ROUGIER ◽  
Robert DUCROC ◽  
Anne-Marie BARLIER-MUR ◽  
...  
Keyword(s):  
Lung Surfactant ◽  
Surfactant Protein ◽  
Mesothelial Cells ◽  
Surfactant Proteins ◽  
Two Dimensional ◽  
Western Blots ◽  
Mrna Species ◽  
Visceral Peritoneum ◽  
Rat Mesentery

Human peritoneal dialysis effluent (PDE) contains a phosphatidylcholine-rich compound similar to the surfactant that lines lung alveoli. This material is secreted by mesothelial cells. Lung surfactant is also characterized by four proteins essential to its function. After having long been considered as lung-specific, some of them have been found in gastric and intestinal epithelial cells. To explore further the similarity between lung and peritoneal surfactants, we investigated whether mesothelial cells also produce surfactant proteins. We used rat transparent mesentery, human visceral peritoneum biopsies and PDE. Surfactant proteins were searched for after one- and two-dimensional SDS/PAGE and Western blotting. On a one-dimensional Western blot, bands at 38 and 66 kDa in rat mesentery, and at 38 and 66 kDa in human peritoneal mesothelial cells (in vivo and in vitro) and PDE, corresponded to monomeric and dimeric forms of lung surfactant protein A (SP-A). On two-dimensional Western blots, the 32 and 38 kDa spots in mesentery and PDE localized at the acidic pH appropriate to the SP-A monomer's isoelectric point. SP-D was also identified at the same 43 kDa molecular mass as in lung. SP-B was not detected in mesenteric samples. Expression of SP mRNA species was also assessed by reverse transcriptase-PCR, which was performed with specific primers of surfactant protein cDNA sequences. With primers of SP-A and SP-D, DNA fragments of the same size were amplified in lung and mesentery, indicating the presence of SP-A and SP-D mRNA species. These fragments were labelled by appropriate probes in a Southern blot. No amplification was obtained for SP-B. These results show that mesentery cells produce SP-A and SP-D, although they are of embryonic origin (mesodermal) and are different from those of the lung and digestive tract (endodermal) that secrete these surfactants.

Strategies for analysis of gene expression: pulmonary surfactant proteins

1990 ◽  
Vol 259 (4) ◽  
pp. L185-L197
Author(s):  
B. R. Stripp ◽  
J. A. Whitsett ◽  
D. L. Lattier
Keyword(s):  
Gene Expression ◽  
Dna Sequences ◽  
Pulmonary Surfactant ◽  
Transcriptional Control ◽  
Lung Surfactant ◽  
Surfactant Protein ◽  
Biologically Active ◽  
Surfactant Proteins ◽  
Regulatory Sequences ◽  
Control Mechanisms

Gene transcription is regulated by the formation of protein-DNA complexes that influence the rate of specific initiation of transcription by RNA polymerase. Recent experimental advances allowing the identification of cis regulatory sequences that specify the binding of trans acting protein factors have made significant contributions to our understanding of the mechanistic complexities of transcriptional regulation. These methodologies have prompted the use of similar strategies to elucidate transcriptional control mechanisms involved in the tissue specific and developmental regulation of pulmonary surfactant protein gene expression. The purpose of this review is to describe various methodologies by which molecular biologists identify and subsequently assay regions of nucleic acids presumed to be integral in gene regulation at the level of transcription. It is well established that genes encoding surfactant proteins are subject to regulation by hormones, cytokines, and a variety of biologically active reagents. Perhaps future studies utilizing molecular tools outlined in this review will be valuable in identification of DNA sequences and protein factors required for the regulation of lung surfactant genes.

Interaction of transferrin saturated with iron with lung surfactant in respiratory failure

1994 ◽  
Vol 77 (2) ◽  
pp. 757-766 ◽  
Author(s):  
M. Hallman ◽  
A. Sarnesto ◽  
K. Bry
Keyword(s):  
Respiratory Failure ◽  
Surface Activity ◽  
Lung Surfactant ◽  
Protein A ◽  
Surfactant Protein ◽  
Lung Damage ◽  
Epithelial Lining ◽  
Surface Absorption ◽  
Epithelial Lining Fluid

Proteins that decrease the surface activity of surfactant accumulate in epithelial lining fluid in respiratory failure. The aim of this study was to isolate a surfactant inhibitor from the airways of rabbits in acute respiratory failure induced by bronchoalveolar lavage (BAL). This inhibitor was identified as being transferrin (TF). Unlike serum TF, TF recovered in respiratory failure was saturated with iron (Fe(3+)-TF). Fe(3+)-TF decreased the surface activity of normal surfactant in vitro, whereas iron-free TF had no effect. In the presence of H2O2 and a reducing agent, Fe(+3)-TF inactivated the surfactant complex: the surface absorption rate was decreased, immunoreactive surfactant protein A was decreased, and malondialdehyde was formed. The acute effects of Fe(3+)-TF and iron-free TF applied to the airways were studied in animal models. In respiratory failure induced by BAL, Fe(3+)-TF deteriorated respiratory failure, whereas iron-free TF had no effect. In respiratory failure induced by hyperoxia for 48 h, administration of iron-free TF ameliorated the respiratory failure and improved the surface activity in BAL. We propose that Fe(3+)-TF accumulating in epithelial lining fluid during lung damage contributes to surfactant inhibition and promotes the formation of free radicals that inactivate the surfactant system.

Lipid composition greatly affects the in vitro surface activity of lung surfactant protein mimics

2007 ◽  
Vol 57 (1) ◽  
pp. 37-55 ◽  
Author(s):  
Shannon L. Seurynck-Servoss ◽  
Nathan J. Brown ◽  
Michelle T. Dohm ◽  
Cindy W. Wu ◽  
Annelise E. Barron
Keyword(s):  
Surface Activity ◽  
Lipid Composition ◽  
Lung Surfactant ◽  
Surfactant Protein ◽  
Protein Mimics ◽  
Lung Surfactant Protein

Mechanisms of peroxynitrite-induced injury to pulmonary surfactants

1993 ◽  
Vol 265 (6) ◽  
pp. L555-L564 ◽  
Author(s):  
I. Y. Haddad ◽  
H. Ischiropoulos ◽  
B. A. Holm ◽  
J. S. Beckman ◽  
J. R. Baker ◽  
...  
Keyword(s):  
Surface Tension ◽  
Dynamic Compression ◽  
Lung Surfactant ◽  
Thiobarbituric Acid ◽  
Surfactant Protein ◽  
Surfactant Proteins ◽  
Minimum Surface ◽  
Surfactant Protein B ◽  
Minimum Surface Tension ◽  
By Products

Activated alveolar macrophages secrete both nitric oxide and superoxide in the alveolar lining fluid which combine rapidly to form peroxynitrite, a potent oxidizing agent capable of damaging lipids and proteins in biological membranes. Peroxynitrite (1 mM) plus 100 microM Fe3+EDTA inhibited calf lung surfactant extract (CLSE) from reaching a minimum surface tension below 10 mN/m on dynamic compression. Peroxynitrite and its by-products reacted with the unsaturated lipid components of CLSE, as evidenced by the appearance of conjugated dienes and thiobarbituric acid products, and damaged all surfactant proteins. A mixture of the hydrophobic proteins [surfactant protein B (SP-B) and surfactant protein C (SP-C)] exposed to peroxynitrite became incapable of lowering phospholipid minimum surface tension on dynamic compression. Exposure of SP-A to peroxynitrite decreased its ability to cause lipid aggregation and to act synergistically with SP-B and SP-C in lowering surface tension of surfactant lipids. Western blot analysis of SP-A exposed to peroxynitrite was consistent with fragmentation and polymerization of the 28- to 36-kDa triplet band, and amino acid analysis revealed the presence of significant levels of 3-nitro-L-tyrosine. We conclude that peroxynitrite and its reactive intermediates inhibit pulmonary surfactant function by lipid peroxidation and damaging surfactant proteins.

Total extracellular surfactant is increased but abnormal in a rat model of gram-negative bacterial pneumonia

2002 ◽  
Vol 283 (3) ◽  
pp. L655-L663 ◽  
Author(s):  
Thomas A. Russo ◽  
Lori A. Bartholomew ◽  
Bruce A. Davidson ◽  
Jadwiga D. Helinski ◽  
Ulrike B. Carlino ◽  
...  
Keyword(s):  
Rat Model ◽  
Lung Surfactant ◽  
Surfactant Protein ◽  
Phospholipid Content ◽  
Aggregate Fraction ◽  
Protein Levels ◽  
Pulmonary Damage ◽  
Aggregate Fractions

An in vivo rat model was used to evaluate the effects of Escherichia coli pneumonia on lung function and surfactant in bronchoalveolar lavage (BAL). Total extracellular surfactant was increased in infected rats compared with controls. BAL phospholipid content in infected rats correlated with the severity of alveolar-capillary leak as reflected in lavage protein levels ( R2= 0.908, P < 0.0001). Western blotting showed that levels of surfactant protein (SP)-A and SP-D in BAL were significantly increased in both large and small aggregate fractions at 2 and 6 h postinstillation of E. coli. SP-B was also increased at these times in the large aggregate fraction of BAL, whereas SP-C levels were increased at 2 h and decreased at 6 h relative to controls. The small-to-large (S/L) aggregate ratio (a marker inversely proportional to surfactant function) was increased in infected rats with >50 mg total BAL protein. There was a significant correlation ( R2= 0.885, P < 0.0001) between increasing S/L ratio in BAL and pulmonary damage assessed by total protein. Pulmonary volumes, compliance, and oxygen exchange were significantly decreased in infected rats with >50 mg of total BAL protein, consistent with surfactant dysfunction. In vitro surface cycling studies with calf lung surfactant extract suggested that bacterially derived factors may have contributed in part to the surfactant alterations seen in vivo.

Erythropoietin Prevents Dialysis Fluid-Induced Apoptosis of Mesothelial Cells

2008 ◽  
Vol 28 (6) ◽  
pp. 648-654 ◽  
Author(s):  
Marina Vorobiov ◽  
Myriam Malki ◽  
Alla Shnaider ◽  
Ana Basok ◽  
Boris Rogachev ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Specific Inhibitor ◽  
Mesothelial Cells ◽  
Short Exposure ◽  
Peritoneal Fibrosis ◽  
Peritoneal Mesothelial Cells ◽  
Visceral Peritoneum ◽  
Induced Apoptosis ◽  
Active Caspase

Background In peritoneal dialysis (PD)-treated patients, denudation of the mesothelium correlates with peritoneal fibrosis and vascular changes. Since recombinant human erythropoietin (rHuEPO) induces a range of cytoprotective cellular responses, rHuEPO treatment may reduce PD fluid (PDF)-induced damage. Methods To investigate the antiapoptotic effect and mechanism of rHuEPO in peritoneal mesothelial cells (PMCs), isolated mice PMCs were used for in vitro characterization of rHuEPO effects. To confirm the in vitro effects, active caspase-3 was analyzed in imprints of liver visceral peritoneum of mice pretreated overnight with rHuEPO (5000 U/kg intraperitoneally) and exposed to PDF (Dianeal 4.25%; Baxter Healthcare, Deerfield, Illinois, USA) for 4 hours. Results Mouse PMCs expressed EPO-receptor mRNA and protein. Short exposure to rHuEPO (5 U/mL) induced phosphorylation of JAK2, STAT5, and ERK1/2. PMCs pretreated for 1 hour with rHuEPO showed reduced PDF-induced caspase-3 activation (49.6%) and DNA fragmentation (38.4%) in comparison to cells treated by PDF alone ( p < 0.05). rHuEPO treatment induced an increase in ERK1/2 phosphorylation and reduced levels of PDF-induced phospho-P38. PD98059, a specific inhibitor of ERK activation, fully blocked the protective effect of rHuEPO. In mice, rHuEPO reduced the apoptotic effect of PDF, as assessed by the level of active caspase-3. Conclusions Our study presents new insights into clinical use of rHuEPO in the setting of PD. We found that rHuEPO provides ERK1/2-dependent protection to PMCs from PDF-induced apoptosis. The use of rHuEPO, or any of its new derivatives that do not stimulate erythropoiesis, should be considered for peritoneal preservation.

Effects of TNF-alpha and phorbol ester on human surfactant protein and MnSOD gene transcription in vitro

1992 ◽  
Vol 262 (6) ◽  
pp. L688-L693 ◽  
Author(s):  
J. A. Whitsett ◽  
J. C. Clark ◽  
J. R. Wispe ◽  
G. S. Pryhuber
Keyword(s):  
Gene Transcription ◽  
Phorbol Ester ◽  
Time Course ◽  
Surfactant Protein ◽  
Surfactant Proteins ◽  
Tnf Alpha ◽  
Inhibitory Effects ◽  
Necrosis Factor Alpha ◽  
Mnsod Gene

Tumor necrosis factor-alpha (TNF-alpha) and the phorbol ester, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) decrease the synthesis of surfactant proteins association with decreased SP-A and SP-B mRNA. Nuclear run-on assays were utilized to test whether the inhibitory effects of TNF-alpha and TPA were associated with changes in surfactant protein gene transcription. SP-A gene transcription was inhibited by both TNF-alpha and TPA as assessed by nuclear run-on assays. Inhibitory effects of both agents on SP-A gene transcription were readily detected within 6 h after exposure and persisted for 24 h. While TNF-alpha and TPA decreased cellular SP-B mRNA content, transcription of the SP-B gene was not influenced by these agents. In contrast to the inhibitory effects of TPA and TNF-alpha on SP-A and SP-B mRNAs, steady-state mRNA and rate of transcription of human manganese superoxide dismutase (MnSOD) were increased by both agents. The time course and extent of increased MnSOD gene transcription by TNF-alpha and TPA were distinct. Transcription of the human beta-actin gene was not altered by either agent. The inhibitory effects of TPA and TNF-alpha on SP-A expression in pulmonary adenocarcinoma cells are associated with the inhibition of SP-A gene transcription. Loss of SP-B mRNA was not accompanied by decreased SP-B gene transcription. Actinomycin D blocked the inhibitory effects of TNF-alpha and TPA on SP-A and SP-B mRNA, supporting a role for posttranscriptional events in the modulation of the expression of the surfactant proteins SP-A and SP-B.

scholarly journals Immunodetection of surfactant proteins in human organ of Corti, Eustachian tube and kidney.

2003 ◽  
Vol 50 (4) ◽  
pp. 1057-1064 ◽  
Author(s):  
Orhan Kankavi
Keyword(s):  
Eustachian Tube ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Organ Of Corti ◽  
Surfactant Protein ◽  
Surfactant Proteins ◽  
Surfactant Protein D ◽  
Western Blots ◽  
Human Organ ◽  
One Dimensional

The presence of surfactant proteins was investigated in the human organ of Corti, Eustachian tube and kidney tissues. It has previously been shown that lamellar bodies are present in hairy cells of organ of Corti, in the cytoplasm of secretory and lumen of tubal glands of Eustachian tube and kidney renal basement membrane. No evidence for the presence of surfactant proteins in the organ of Corti and kidney has been presented until now. The aim of this study was to find out if surfactant proteins were expressed in other epithelia such as organ of Corti, Eustachian tube and kidney. Surfactant proteins were identified using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. On one-dimensional Western blots, bands for surfactant protein A in human Eustachian tube (SP-A, 34 kDa) and in kidney extracts, and for surfactant protein D (SP-D, 43 kDa) in Eustachian tube and in kidney extracts (SP-D, 86 kDa), and for surfactant protein B (SP-B, 8 kDa) in human Eustachian tube and organ of Corti extracts were detected. Bands corresponded to monomeric forms of lung surfactant proteins. These results indicate the presence of SP-A and SP-D in kidney epithelium, SP-A, SP-B and SP-D in Eustachian tube and SP-B in the organ of Corti.

Adsorption of fetal surfactant protein SP-B on the human amnion at term and on amniocytes incubated with fetal surfactant in vitro

1991 ◽  
Vol 3 (4) ◽  
pp. 421 ◽  
Author(s):  
GE Newman ◽  
PJ Phizackerley ◽  
Bernal A Lopez ◽  
GR Noble ◽  
AC Willis
Keyword(s):  
Monoclonal Antibody ◽  
Amniotic Fluid ◽  
Human Lung ◽  
Lung Surfactant ◽  
Surfactant Protein ◽  
Post Mortem ◽  
Human Amniotic Fluid ◽  
Human Amnion ◽  
Immunohistochemical Methods

Fetal surfactant stimulates the synthesis of prostaglandins by slices of human amnion at term and by a human amnion cell line, and these effects are partly dependent upon surfactant apoproteins. In this paper, methods are described for the purification of surfactant from human amniotic fluid and from post-mortem human lung. A procedure is described for the purification of surfactant protein SP-B from human amniotic fluid, and the sequence of 20 amino acids at the N-terminal has been determined. A monoclonal antibody generated against human lung surfactant has been shown to react with SP-B from amniotic-fluid surfactant, and the presence of SP-B on the surface of the amnion at term has been demonstrated by immunohistochemical methods. It has also been shown that SP-B from surfactant is present on the surface of amniocytes incubated with surfactant in vitro.

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