Mol. Hum. Reprod. Advance Access originally published online on April 20, 2007
Molecular Human Reproduction 2007 13(6):409-418; doi:10.1093/molehr/gam025
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Evidence of in vitro differential secretion of 72 and 92 kDa type IV collagenases after selective exposure to lipopolysaccharide in human fetal membranes
1 Direction of Research, Instituto Nacional de Perinatologia ‘Isidro Espinosa de los Reyes’, Mexico City 11000, Mexico 2 Biomedical Research Institute, Universidad Nacional Autónoma de México, Mexico City, 04510, Mexico 3 Neonatology Branch, Instituto Nacional de Perinatologia ‘Isidro Espinosa de los Reyes’, Mexico City 11000, Mexico 4 Obstetrics and Gynecology Branch, Instituto Nacional de Perinatologia ‘Isidro Espinosa de los Reyes’, Mexico City 11000, Mexico 5 Microbiology and Parasitology Branch, Instituto Nacional de Perinatologia ‘Isidro Espinosa de los Reyes’, Mexico City 11000, Mexico 6 Biomedical Research Branch, Instituto Nacional de Perinatologia ‘Isidro Espinosa de los Reyes’, Mexico City 11000, Mexico 7 Obstetrics and Gynecology Branch, Instituto Nacional de Perinatologia ‘Isidro Espinosa de los Reyes’, Mexico City 11000, Mexico 8 Research Unit, Instituto de Oftalmología ‘Conde de Valenciana’ Mexico City, 06800, Mexico
9 Corresponding address. Tel/Fax: +52-55-5520-0034; E-mail: zaga{at}universo.com
Premature rupture of chorioamniotic membranes complicated with intrauterine infection has been associated to degradation of extracellular matrix (ECM), which could explain local morphological changes. We used a culture system in which the chorioamniotic membranes form two independent chambers, allowing for the selective stimulation of either the amnion (AMN) and/or the choriodecidua (CHD) regions. Lipopolysaccharide (500 ng/ml) was added to the AMN and/or the CHD; secretions and gelatinolytic activity of matrix metalloproteinase (MMP)-2 and MMP-9 were measured in both compartments by enzyme-linked immunosorbent assay (ELISA) and zymography. Secretions of TIMP-1, TIMP-2 and TIMP-4 were measured by ELISA. Both metalloproteinases were immunolocalized in tissue sections. All stimulation modalities induced a similar proMMP-2 and proMMP-9 secretion pattern in the CHD with concentrations of 2.49 ng/ml and 90.91 pg/ml, respectively; the AMN showed no significant changes. The active forms of both enzymes did not change with any stimulation modality. TIMP-1, TIMP-2 and TIMP-4 secretions remained without significant changes (P = 0.41). ECM degradation and structural disarrangement were evident after stimulation. Secretion of proMMP-2 and proMMP-9 mainly in the CHD, presence of active forms associated to the tissue and minor changes in TIMPs secretion could favor ECM degradation and explain the weakening and thinning associated with the pathological rupture of chorioamniotic membranes.
Key words: intrauterine infection/chorioamniotic membranes/matrix metalloproteinases/extracellular matrix
Submitted on December 18, 2007; resubmitted on February 14, 2007; accepted on March 9, 2007.