Interleukin-10 inhibition of gelatinases in fetal membranes: therapeutic implications in preterm premature rupture of membranes.
OBJECTIVE
To examine the effect of interleukin-10 on production and regulation of gelatinases by amniochorion in an in vitro model of infection.


METHODS
We placed amniochorionic membranes collected from eight women who had elective repeat cesareans at term in an organ explant culture system. After 48 hours in culture, the membranes were stimulated with lipopolysaccharide (50 ng/mL), and some were costimulated with interleukin-10 (500 ng/mL). Tissue and media samples were collected after 24-hour stimulation. Quantitative polymerase chain reactions and enzyme-linked immunosorbent assays were used to evaluate matrix metalloproteinase 2 and matrix metalloproteinase 9 messenger RNA and proteins, respectively.


RESULTS
Lipopolysaccharide stimulation induced 55.14 transcripts of matrix metalloproteinase 9, compared with 0.83 in control tissues (P <.001). Costimulation with interleukin-10 and lipopolysaccharide significantly reduced matrix metalloproteinase 9 messenger RNA levels to 10 transcripts (P <.001). Lipopolysaccharide stimulation produced 29.25 ng/mL of immunoreactive matrix metalloproteinase 9, which was reduced to 6.3 ng/mL (P(adj) =.016) after costimulation with interleukin-10. Although not significant, matrix metalloproteinase 2 messenger RNA levels were higher in lipopolysaccharide-stimulated tissues (4.37 x 10(6) transcripts) compared with control (2.8 x 10(5) transcripts; P(adj) =.08), with a significant decrease in matrix metalloproteinase 2 messenger RNA levels in interleukin-10- costimulated tissues (2.9 x 10(6); P(adj) =.007). Interleukin-10 costimulation resulted in a significant decrease in matrix metalloproteinase 2 protein production (203.1 [lipopolysaccharide] and 149.75 [with interleukin-10]; P(adj) <.001).


CONCLUSIONS
Interleukin-10 eliminated lipopolysaccharide induction of matrix metalloproteinase 2 and 9 in amniochorion.
Fortunato
S J
SJ
The Perinatal Research Center of the Women's Health Research and Education Foundation, The Maternal-Fetal Group and Aquinas College, Department of Preventive Medicine, Vanderbilt University Medical Center, Nashville, Tennessee 37203, USA. fortunat@edge.net
Menon
R
R
Lombardi
S J
SJ
LaFleur
B
B
D000650:Amnion / Q000201:enzymology*; D002821:Chorioamnionitis / Q000201:enzymology; D002823:Chorion / Q000201:enzymology*; D046508:Culture Techniques; D004790:Enzyme Induction; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D005260:Female; D005322:Premature Rupture of Fetal Membranes / Q000201:enzymology* / Q000517:prevention & control; D006801:Humans; D016753:Interleukin-10 / Q000494:pharmacology* / Q000627:therapeutic use; D008070:Lipopolysaccharides / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis* / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis* / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000201:enzymology; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D008070:Lipopolysaccharides; D061965:Matrix Metalloproteinase Inhibitors; D012333:RNA, Messenger; D011994:Recombinant Proteins; D016753:Interleukin-10; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9