Interleukin-1beta-induced prostaglandin E2 production in human myometrial cells: role of a pertussis toxin-sensitive component.
OBJECTIVE
The objective of this study was to evaluate the possible role of pertussis toxin (PTX)-sensitive G-protein(s) in interleukin-1beta (IL-1) signaling in human myometrial cells (HMC).


METHODS
Primary cultures of HMC were stimulated with human recombinant IL-1 alone or in combination with PTX. Prostaglandin (PG) E2 in the medium was measured by radioimmunoassay, cyclooxygenase type 2 (Cox-2) and IkappaB by western analysis, and the activities of two members of the mitogen-activated protein kinase (MAPK) family of enzymes, ERK-2 and JNK, by the phosphorylation of appropriate substrates.


RESULTS
IL-1 increased PGE2 output during an 18-hr long incubation by 21.7-fold (n = 5 experiments). This increase was inhibited by 57% after pretreatment overnight with PTX. IL-1-induced expression of Cox-2 protein was also suppressed to a similar degree in PTX-treated HMC cultures. Degradation of the nuclear factor kappa B (NF-kappaB)-inhibiting protein (IkappaB), a critical step in IL-1 signaling to the nucleus, was significantly inhibited by PTX, as was IL-1-induced activation of ERK-2 and JNK.


CONCLUSIONS
It is suggested that the occupied IL-1 receptor-generated signal in HMC is transmitted by multiple pathways. One is coupled to a PTX-sensitive G-protein upstream from the MAPK phosphorylation cascade. This, in turn, may interact with another signaling pathway, the activation of NF-kappaB, via the phosphorylation of the IkappaB kinase complex.
Hertelendy
F
F
Department of Obstetrics, Gynecology and Women's Health, St Louis University School of Medicine and St Mary's Health Center, Missouri 63117, USA. hertelf@slu.edu
Rastogi
P
P
Molnár
M
M
Romero
R
R
D002478:Cells, Cultured; D015232:Dinoprostone / Q000096:biosynthesis*; D005260:Female; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D008954:Models, Biological; D009215:Myometrium / Q000166:cytology / Q000187:drug effects*; D037342:Pertussis Toxin; D017697:Premenopause; D015398:Signal Transduction; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D007375:Interleukin-1; D010566:Virulence Factors, Bordetella; D037342:Pertussis Toxin; D015232:Dinoprostone