Mechanisms of abruption-induced premature rupture of the fetal membranes: Thrombin enhanced decidual matrix metalloproteinase-3 (stromelysin-1) expression.
OBJECTIVE
The aim of the study was to evaluate thrombin and progestin effects on matrix metalloproteinase-3 expression in term decidual cells as a mechanism of abruption-related preterm delivery.


METHODS
Decidual cells were isolated by standard techniques, purified to homogeneity, grown to confluence, and passaged. Cultures were primed with 10 (-8) M estradiol or estradiol plus 10 (-7) progestin and then incubated in a defined medium with corresponding steroid(s) plus or minus thrombin or the protease-activated thrombin receptor-1 agonist for 24 hours. Secreted matrix metalloproteinase-3 levels were assessed by enzyme-linked immunosorbent assay, and immunoblotting and messenger RNA levels were measured by Northern blotting and quantitative reverse transcription-polymerase chain reaction.


RESULTS
Immunoreactive matrix metalloproteinase-3 levels were inhibited 66% by estradiol plus progestin versus estradiol ( P < .05). Thrombin elicited a dose-dependent reversal in this progestin inhibition, producing a 2.5-fold increase at 2.5 U/mL ( P < .05) that attained 33% of matrix metalloproteinase-3 levels in parallel incubations with estradiol plus thrombin. Protease-activated thrombin receptor-1 agonist mimicked 60% of thrombin-enhanced matrix metalloproteinase-3 output. Immunoblotting validated the enzyme-linked immunosorbent assay results. Northern blotting and quantitative reverse transcription-polymerase chain reaction demonstrated corresponding effects on steady-state messenger RNA levels.


CONCLUSIONS
Abruption-generated thrombin promotes preterm delivery by mediating fetal membrane extracellular matrix degradation via enhanced decidual cell matrix metalloproteinase-3 expression, whereas progesterone blunts this thrombin-induced effect.
Mackenzie
Andrew P
AP
Department of Obstetrics and Gynecology, New York University School of Medicine, New York, NY, USA.
Schatz
Frederick
F
Krikun
Graciela
G
Funai
Edmund F
EF
Kadner
Susan
S
Lockwood
Charles J
CJ
D001483:Base Sequence; D015152:Blotting, Northern; D002478:Cells, Cultured; D003656:Decidua / Q000166:cytology / Q000187:drug effects; D005321:Extraembryonic Membranes / Q000166:cytology / Q000187:drug effects; D005260:Female; D005322:Premature Rupture of Fetal Membranes / Q000209:etiology; D006801:Humans; D066298:In Vitro Techniques; D019278:Matrix Metalloproteinase 3 / Q000187:drug effects* / Q000378:metabolism*; D008969:Molecular Sequence Data; D011247:Pregnancy; D011372:Progestins / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013917:Thrombin / Q000494:pharmacology*
D011372:Progestins; D012333:RNA, Messenger; D013917:Thrombin; D019278:Matrix Metalloproteinase 3