Serine proteinase inhibitors influence the stability of tropoelastin mRNA in neonatal rat lung fibroblast cultures.
Elastin, an elastic extracellular structural protein, is a polymer comprised of soluble tropoelastin (TE) monomers that are joined by covalent cross-links and become insoluble. In cultured vascular smooth muscle cells, the steady-state level of TE mRNA is influenced by soluble elastin moieties in the culture medium, either TE or its fragmentation products. We have hypothesized that an enzyme-mediated proteolytic event may modulate the quantities of TE and its fragmentation products in the culture medium of mesenchymal cells, and thereby indirectly regulate the steady-state level of TE mRNA. Neonatal rat lung fibroblasts were cultured in the presence or absence of the serine proteinase inhibitor, aprotinin, and the quantities of soluble elastin and TE mRNA were analyzed. Exposures to aprotinin lasting up to 12 h increased the soluble elastin content of the culture medium. The increase in the soluble elastin content did not reflect an increase in TE mRNA, which diminished after exposures for 12 h or longer. The decrease in TE mRNA resulted from a decrease in its half-life, rather than a decrease in the rate of TE gene transcription. Aprotinin did not reduce TE mRNA in plasminogen-depleted cultures, but the effect of aprotinin was evident when purified plasminogen was added back to the cultures. Therefore, a serine proteinase, possibly plasmin, may participate in a feedback mechanism and modulate the quantity of TE in lung fibroblast cultures. This mechanism may help ensure that intracellular TE synthesis occurs in tandem with extracellular elastin deposition and cross-linking.
McGowan
S E
SE
Department of Veterans Affairs Research Service, Iowa City, Iowa 52242, USA.
Liu
R
R
Harvey
C S
CS
Jaeckel
E C
EC
D000818:Animals; D000831:Animals, Newborn; D007611:Aprotinin / Q000494:pharmacology*; D015152:Blotting, Northern; D002478:Cells, Cultured; D004549:Elastin / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D005341:Fibrinolysin; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D007700:Kinetics; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects*; D014333:Tropoelastin / Q000096:biosynthesis*
D010446:Peptide Fragments; D012333:RNA, Messenger; D015842:Serine Proteinase Inhibitors; D014333:Tropoelastin; D004549:Elastin; D007611:Aprotinin; D005341:Fibrinolysin