Pigment epithelial-derived factor in human fetal membranes.

OBJECTIVE: Our main objective was to document, pigment epithelial-derived factor (PEDF), a secreted serine protease inhibitor with anti-angiogenic, anti-inflammatory, and anti-oxidant properties, expression in human fetal membranes from preterm prelabor rupture of the membranes (pPROM) and in in vitro cultures stimulated with cigarette smoke extract (CSE) or lipopolysaccharides (LPS), two major risk factors for pPROM (behavioral and bacterial, respectively). METHOD: We documented PEDF mRNA expression in clinical samples of fetal membranes from patients with pPROM using quantitative RT-PCR. Also, mRNA and protein levels were documented in fetal membranes (from normal term cesarean sections [not in labor]) in an organ explant system stimulated with CSE or lipopolysaccharide (LPS). Immunohistochemistry (IHC) was used to localize PEDF in fetal membranes. RESULTS: We report no changes in PEDF mRNA expression in pPROM compared to term births (p = .59) or after treatment with CSE or LPS. However, by adding sulforaphane the PEDF mRNA expression increased significantly p < .000032. PEDF was localized to both amnion and chorion layers, but no difference was seen in staining intensities after CSE or LPS treatment compared to control. CONCLUSIONS: PEDF, a product of fetal membrane cells, is unaltered in pPROM or after exposure to risk factors of pPROM. The antioxidant stimulating substance sulforaphane contribute to an increase in PEDF mRNA in fetal membranes.

Anti-inflammatory Elafin in human fetal membranes.

OBJECTIVE: Elafin is a low molecular weight protein with antileukoproteinase, anti-inflammatory, antibacterial and immunomodulating properties. The profile of Elafin in fetal membranes is not well characterized. This study determined the changes in Elafin expression and concentration in human fetal membrane from patients with preterm prelabor rupture of membranes (PPROM) and in vitro in response to intra-amniotic polymicrobial pathogens. METHOD: Elafin messenger RNA (mRNA) expressions were studied in fetal membranes from PPROM, normal term as well as in normal term not in labor membranes in an organ explant system treated (24 h) with lipopolysaccharide (LPS), using quantitative reverse transcription-polymerase chain reaction (RT-PCR). Enzyme-linked immunosorbent assay (ELISA) measured Elafin concentrations in culture supernatants from tissues treated with LPS and polybacterial combinations of heat-inactivated Mycoplasma hominis (MH), Ureaplasma urealyticum (UU) and Gardnerella vaginalis (GV). RESULTS: Elafin mRNA expression in fetal membranes from women with PPROM was significantly higher compared to women who delivered at term after normal pregnancy (5.09±3.50 vs. 11.71±2.21; P<0.05). In vitro, LPS-stimulated membranes showed a significantly increased Elafin m-RNA expression (P<0.05). However, the protein levels after LPS stimulation was not changed. Similarly, polymicrobial-treated fetal membranes also showed no changes in Elafin protein concentrations compared to untreated controls. CONCLUSION: Higher Elafin expression in PPROM fetal membranes suggests a host response to an inflammatory pathology. However, lack of Elafin response to LPS and polymicrobial treatment is indicative of the minimal anti-inflammatory impact of this molecule in fetal membranes.