title: Hyperoxia alone causes changes in lung proteoglycans and hyaluronan in neonatal rat pups.
abstract: Specific changes in composition and content of lung extracellular matrix (ECM) proteoglycans (PGs) and hyaluronan (HA) have been observed during the acute response to damage in several forms of injury including infant respiratory distress syndrome (IRDS). These ECM components are thought to modulate the healing response. Hyperoxia, a contributing factor to IRDS, is known to damage both adult and developing lung, however, the extent and pattern of impairment depends on lung maturity. We hypothesized that exposing neonatal rats to hyperoxia alone might result in changes in lung HA, as well as in age-specific changes in lung PGs, similar to those shown to occur in IRDS. In control rats, lung HA decreased over the first 10 days of life, whereas rats exposed to hyperoxia exhibited a time-dependent, time-limited increase in both lung HA and lung wet weight. Histochemistry showed the HA in hyperoxia-exposed lungs to be accumulated in perivascular cuffs of medium sized arteries, and in the alveolar walls. Rats were then exposed to normoxia or hyperoxia for 7 days beginning at either 3 days of life (neonatal) or 21 days (adolescent), and lung tissue was cultured in the presence of [35S]-sulfate to label newly synthesized PGs. Proteoglycans were extracted, and analyzed by isopycnic CsCl gradient centrifugation, sequential enzymatic deglycosylation, size chromatography, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). When controlled for total protein extracted, 63% more label was incorporated into large molecular weight material in the tissue exposed to hyperoxia, with a 95% increase in incorporation in the most dense fraction, D1. [35S]-Sulfate incorporation into chondroitin and dermatan sulfate in hyperoxic tissue specifically increased 116% (242% in the D1 fraction), while incorporation into heparan sulfate remained essentially unchanged. There was a nearly fivefold increase in [35S]-sulfate incorporation into chondroitin sulfate chains in the D1 fraction. When the D1 fractions of extracts of treated and control rat lungs were compared on SDS-PAGE, a large chondroitin sulfate proteoglycan (CSPG; core protein of 195 kDa) was upregulated in the D1 fraction from hyperoxic tissue of neonatal rats, but was not detected in the lungs of adolescent animals exposed to hyperoxia. This CSPG and four additional large CSPGs were noted to be upregulated on western blotting by a polyclonal antibody directed against the G1 domain of the aggrecan protein core. We conclude that hyperoxia alone causes an increase in lung HA and lung water, and speculate that this contributes significantly to the clinical syndrome of IRDS. In addition, several large CSPGs are upregulated by hyperoxic exposure in a developmentally specific manner. We speculate that this increase in CSPGs may interfere with the normal developmental sequence of events, contributing to hypoalveolarization.
authors.affiliation: Department of Pediatrics, University of Florida, Gainesville, USA.
authors.forename: S E
authors.initials: SE
authors.lastname: Juul
authors.forename: R C
authors.initials: RC
authors.lastname: Krueger
authors.forename: L
authors.initials: L
authors.lastname: Scofield
authors.forename: M B
authors.initials: MB
authors.lastname: Hershenson
authors.forename: N B
authors.initials: NB
authors.lastname: Schwartz
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D002458:Cell Fractionation; D002586:Cesium; D002712:Chlorides; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006026:Glycoside Hydrolases; D006820:Hyaluronic Acid / Q000378:metabolism*; D018496:Hyperoxia / Q000378:metabolism*; D008168:Lung / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism*; D011650:Pulmonary Alveoli / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013462:Sulfur Radioisotopes
chemical_list: D002712:Chlorides; D011509:Proteoglycans; D013462:Sulfur Radioisotopes; D002586:Cesium; D006820:Hyaluronic Acid; D006026:Glycoside Hydrolases; C028019:cesium chloride