Ramkumar Menon, Stefan Gerber, Stephen J. Fortunato, Steven S. Witkin
Lipopolysaccharide stimulation of 70 kilo Dalton heat shock protein messenger ribonucleic acid production in cultured human fetal membranes
Objective: The 70 kilo Dalton heat shock protein is upregulated
when cells are under physiological stress. It
prevents protein denaturation and incorrect polypeptide
assembly, and inhibits apoptosis as well as the transcription of genes coding for pro-inflammatory cytokines. To
evaluate if up-regulation of heat shock protein 70 can
occur during pregnancy, we examined whether addition
of bacterial lipopolysaccharide to human amniochorion
membranes in vitro stimulated heat shock protein 70
gene transcription.
Materials and methods: Amniochorionic membranes
(n = 5), collected at the time of elective repeat cesarean
section prior to labor from normal term gestations, were
placed in an organ explant system. After 48 hour in
culture, the membranes were stimulated with lipopolysaccharide
for 24 hours. Total RNA was extracted and
subjected to an oligo dT primed reverse transcriptase
reaction followed by polymerase chain reaction (PCR)
using heat shock protein 70 specific primers. PCR products
were hybridized with biotinylated internal probes
and identified by enzyme-linked immunosorbent assay
(ELISA). Results were analyzed by Mann-Whitney U
test. A p < 0.05 was significant.
Results: Heat shock protein 70 messenger RNA was expressed
by all fetal membrane preparations both prior to
and following in vitro culture. Addition of lipopolysaccharide
increased the concentrations of heat shock protein
70 messenger RNA in each sample tested from a
mean of 35.5 ± 29.6 ng/milliliter (12.1–80.1 ng/milliliter)
to 169.6 ± 69.9 ng/ml (51.7–218.2 ng/milliliter)
(p = 0.03).
Conclusion: Human fetal membranes constitutively express
heat shock protein 70 messenger ribonucleic acid.
Bacterial lipopolysaccharide markedly stimulated heat
shock protein 70 messenger RNA gene transcription in
human fetal membranes. Thus, heat shock protein 70 is
inducible in fetal membranes and may facilitate fetal
survival under adverse conditions.
Journal of Perinatal Medicine, Walter de Gruyter
Print ISSN: 1619-3997
Volume: 29, 04/2001
Pages: 133 - 136
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