Stromelysins in placental membranes and amniotic fluid with premature rupture of membranes.
OBJECTIVE
To determine the expression and site of production of stromelysins in fetal membranes and to measure stromelysin 1 levels in amniotic fluid and amniochorion culture media.


METHODS
Amniochorionic membranes were cultured from organ explant. Membranes were stimulated with lipopolysaccharide for 24 hours after a 48-hour preincubation period. Membranes were also collected from women after vaginal deliveries. RNA samples from those tissues were subjected to reverse transcriptase-polymerase chain reaction using primers specific for stromelysin 1, stromelysin 2, stromelysin 3, and matrilysin. In situ hybridization and immunohistochemistry were used to localize stromelysin mRNA and peptide. Levels of stromelysin 1 in culture media and amniotic fluid collected from women with preterm premature rupture of membranes (PROM) and at term with intact membranes were compared using enzyme-linked immunosorbant assay.


RESULTS
Amniochorion in culture and from laboring and nonlaboring women expressed all three stromelysins. In situ hybridization showed stromelysin mRNA in amnion, chorion, and extracellular matrix. Immunohistochemical analysis localized stromelysin 1 protein to those same regions. Amniotic fluid levels of stromelysin 1 were higher in preterm PROM amniotic fluids (median 3.2 ng/mL) compared with term deliveries with intact membranes (median 1.3 ng/mL) (P = .02). Lipopolysaccharide stimulation in culture increased the release of stromelysin 1 from fetal membranes compared with control (median 70.35 versus 15.8 ng/mL, respectively, P = .05).


CONCLUSIONS
Human fetal membranes are a source of stromelysins 1, 2, and 3. Increased stromelysin 1 during preterm PROM and in vitro after lipopolysaccharide stimulation suggests a possible effect of that matrix metalloproteinase in PROM.
Fortunato
S J
SJ
Maternal Fetal Group, Perinatal Research Center, Nashville, Tennessee 37203, USA. fortunat@edge.net
Menon
R
R
Lombardi
S J
SJ
D000653:Amniotic Fluid / Q000737:chemistry* / Q000378:metabolism*; D005260:Female; D005322:Premature Rupture of Fetal Membranes / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis* / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D053506:Matrix Metalloproteinase 10; D053507:Matrix Metalloproteinase 11; D019278:Matrix Metalloproteinase 3 / Q000032:analysis* / Q000096:biosynthesis* / Q000235:genetics; D008666:Metalloendopeptidases / Q000032:analysis* / Q000096:biosynthesis* / Q000235:genetics; D010920:Placenta / Q000737:chemistry* / Q000378:metabolism*; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis
D006023:Glycoproteins; D012333:RNA, Messenger; D053507:Matrix Metalloproteinase 11; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D053506:Matrix Metalloproteinase 10