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1 Dipartimento di Biochimica
"A. Castellani,
We extracted and
isolated proteoglycans from lung tissue samples obtained
from three groups of anesthetized rabbits:
1) control animals (C; n = 8) killed by overdose after 180 min;
2) animals receiving an
intravenous saline infusion (S; n = 4, 1.5 ml · kg-1 · min-1)
for 180 min; 3) animals receiving an
intravenous bolus of 200 µg of pancreatic elastase (E;
n = 4), killed after 200 min. The lung
dry weight-to-wet weight ratio in the three groups was 5.2 ± 0.2, 6.0 ± 0.4, and 5.6 ± 0.5, respectively. Gel-filtration analysis
showed a massive fragmentation of the versican family of the
extracellular matrix (ECM) in the S groups and a marked degradation of
heparan sulfate-containing proteoglycans, including perlecan of the
basement membrane, in the E group. The binding properties of total
proteoglycans to other ECM components were lowered in both groups
relative to control. The decrease in proteoglycan binding was more
pronounced for collagen type IV in the E group relative to C
(
93.5%, P < 0.05) and for
hyaluronic acid in the S groups (85.8%,
P < 0.05). These findings suggest
that elastase treatment produces a major degree of damage to the
organization of basement membrane, whereas saline loading affects more
markedly the architecture of interstitial ECM. Qualitative zymography
performed on lung extracts showed increased gelatinase activities in
both S and E groups, providing direct evidence that the activation of
tissue proteinases may play a role in acute lung injury.
extracellular matrix; basement membrane; gelatinase; interstitial fluid dynamics
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