Amniochorion gelatinase-gelatinase inhibitor imbalance in vitro

A possible infectious pathway to rupture

Stephen J. Fortunato, Ramkumar Menon, Salvatore J. Lombardi

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Objective: To estimate the effect of lipopolysaccharide on gelatinases and tissue inhibitors of matrix metalloproteinase 2 (gelatinase inhibitor) balance in human fetal membranes. Methods: Amniochorionic membranes in organ explant were stimulated with 1000 ng/mL lipopolysaccharide for 24 hours after a 48-hour preincubation period. Quantitative competitive polymerase chain reaction (PCR) was done to quantitate messenger RNAs for gelatinase A and B (matrix metalloproteinase 2 and 9) and tissue inhibitor of metalloproteinase 2. Protein levels were assayed by enzyme-linked immunosorbant assay. The molar ratio between gelatinases and tissue inhibitor of metalloproteinase 2 was calculated. Statistical evaluation was done by Mann-Whitney U test.Results: Lipopolysaccharide stimulation produced 3.6 x 106 and 366 transcripts of gelatinase A and B, respectively, compared with only 5.9 x 104 (P = .009) and three transcripts (P = .006), respectively, in the controls. Lipopolysaccharide stimulation released 210 ng/mL compared with 7 ng/mL of gelatinase A and B proteins compared with 120 (P = .01) and 4.6 ng/mL (P = .3) in controls, respectively. Control amniochorion produced 5.7 x 105 transcripts of tissue inhibitor of metalloproteinase 2, whereas lipopolysaccharide stimulation produced 4.1 x 105 transcripts (P = .69). Lipopolysaccharide reduced the release of this inhibitor from 114 ng/mL to 68 ng/mL (P = .007). The molar ratio between gelatinases and tissue inhibitor of metalloproteinase 2 increased from a balanced ratio of 1:1 to 3.1:1 after 1000 ng/mL of lipopolysaccharide.Conclusion: Lipopolysaccharide increased the expression and release of gelatinases and decreased its inhibitor, which shifted the balance in favor of gelatinase activity leading to membrane degradation that predisposes to premature rupture of membranes. Copyright (C) 2000 The American College of Obstetricians and Gynecologists.

Original languageEnglish (US)
Pages (from-to)240-244
Number of pages5
JournalObstetrics and Gynecology
Volume95
Issue number2
DOIs
StatePublished - Feb 2000
Externally publishedYes

Fingerprint

Gelatinases
Matrix Metalloproteinase Inhibitors
Lipopolysaccharides
Rupture
Tissue Inhibitor of Metalloproteinase-2
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Membranes
Extraembryonic Membranes
In Vitro Techniques
Nonparametric Statistics
Proteins
Polymerase Chain Reaction
Messenger RNA
Enzymes

ASJC Scopus subject areas

  • Obstetrics and Gynecology

Cite this

Amniochorion gelatinase-gelatinase inhibitor imbalance in vitro : A possible infectious pathway to rupture. / Fortunato, Stephen J.; Menon, Ramkumar; Lombardi, Salvatore J.

In: Obstetrics and Gynecology, Vol. 95, No. 2, 02.2000, p. 240-244.

Research output: Contribution to journalArticle

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N2 - Objective: To estimate the effect of lipopolysaccharide on gelatinases and tissue inhibitors of matrix metalloproteinase 2 (gelatinase inhibitor) balance in human fetal membranes. Methods: Amniochorionic membranes in organ explant were stimulated with 1000 ng/mL lipopolysaccharide for 24 hours after a 48-hour preincubation period. Quantitative competitive polymerase chain reaction (PCR) was done to quantitate messenger RNAs for gelatinase A and B (matrix metalloproteinase 2 and 9) and tissue inhibitor of metalloproteinase 2. Protein levels were assayed by enzyme-linked immunosorbant assay. The molar ratio between gelatinases and tissue inhibitor of metalloproteinase 2 was calculated. Statistical evaluation was done by Mann-Whitney U test.Results: Lipopolysaccharide stimulation produced 3.6 x 106 and 366 transcripts of gelatinase A and B, respectively, compared with only 5.9 x 104 (P = .009) and three transcripts (P = .006), respectively, in the controls. Lipopolysaccharide stimulation released 210 ng/mL compared with 7 ng/mL of gelatinase A and B proteins compared with 120 (P = .01) and 4.6 ng/mL (P = .3) in controls, respectively. Control amniochorion produced 5.7 x 105 transcripts of tissue inhibitor of metalloproteinase 2, whereas lipopolysaccharide stimulation produced 4.1 x 105 transcripts (P = .69). Lipopolysaccharide reduced the release of this inhibitor from 114 ng/mL to 68 ng/mL (P = .007). The molar ratio between gelatinases and tissue inhibitor of metalloproteinase 2 increased from a balanced ratio of 1:1 to 3.1:1 after 1000 ng/mL of lipopolysaccharide.Conclusion: Lipopolysaccharide increased the expression and release of gelatinases and decreased its inhibitor, which shifted the balance in favor of gelatinase activity leading to membrane degradation that predisposes to premature rupture of membranes. Copyright (C) 2000 The American College of Obstetricians and Gynecologists.

AB - Objective: To estimate the effect of lipopolysaccharide on gelatinases and tissue inhibitors of matrix metalloproteinase 2 (gelatinase inhibitor) balance in human fetal membranes. Methods: Amniochorionic membranes in organ explant were stimulated with 1000 ng/mL lipopolysaccharide for 24 hours after a 48-hour preincubation period. Quantitative competitive polymerase chain reaction (PCR) was done to quantitate messenger RNAs for gelatinase A and B (matrix metalloproteinase 2 and 9) and tissue inhibitor of metalloproteinase 2. Protein levels were assayed by enzyme-linked immunosorbant assay. The molar ratio between gelatinases and tissue inhibitor of metalloproteinase 2 was calculated. Statistical evaluation was done by Mann-Whitney U test.Results: Lipopolysaccharide stimulation produced 3.6 x 106 and 366 transcripts of gelatinase A and B, respectively, compared with only 5.9 x 104 (P = .009) and three transcripts (P = .006), respectively, in the controls. Lipopolysaccharide stimulation released 210 ng/mL compared with 7 ng/mL of gelatinase A and B proteins compared with 120 (P = .01) and 4.6 ng/mL (P = .3) in controls, respectively. Control amniochorion produced 5.7 x 105 transcripts of tissue inhibitor of metalloproteinase 2, whereas lipopolysaccharide stimulation produced 4.1 x 105 transcripts (P = .69). Lipopolysaccharide reduced the release of this inhibitor from 114 ng/mL to 68 ng/mL (P = .007). The molar ratio between gelatinases and tissue inhibitor of metalloproteinase 2 increased from a balanced ratio of 1:1 to 3.1:1 after 1000 ng/mL of lipopolysaccharide.Conclusion: Lipopolysaccharide increased the expression and release of gelatinases and decreased its inhibitor, which shifted the balance in favor of gelatinase activity leading to membrane degradation that predisposes to premature rupture of membranes. Copyright (C) 2000 The American College of Obstetricians and Gynecologists.

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