Lipopolysaccharide stimulation of trophoblasts induces corticotropin-releasing hormone expression through MyD88.
OBJECTIVE
We hypothesized that intrauterine infection may lead to placental corticotrophin-releasing hormone (CRH) expression via Toll-like receptor signaling.


METHODS
To test this hypothesis JEG3 cells were stimulated with lipopolysaccharide (LPS), chlamydial heat shock protein 60, and interleukin (IL)-1. CRH expression was assessed by reverse transcription polymerase chain reaction (RT-PCR). The signaling mechanisms that were involved were examined in transient transfection experiments with beta-galactosidase, CRH-luciferase, cyclic adenosine monophosphate (AMP) response element-luciferase, dominant-negative (DN)-myeloid differentiation primary response gene (MyD88) and DN-toll-IL-1-receptor domain containing adapter inducing interferon (TRIF) vectors. Luciferase activity was determined by luciferase assay. Beta-galactosidase assay was performed to determine transfection efficiency.


RESULTS
LPS, chlamydial heat shock protein 60, and IL-1 stimulation led to CRH expression in the JEG3 cells. LPS-induced CRH expression was not due to the autocrine effect of LPS-induced IL-1 because the supernatant from LPS-conditioned JEG3 cells did not induce CRH expression in the naïve cells. DN-MyD88, but not DN-TRIF, blocked the LPS-induced CRH expression. The cAMP response element did not play a role in LPS-induced CRH expression.


CONCLUSIONS
Toll-like receptor signaling 4 may induce placental CRH expression through MyD88.
Uh
Andy
A
Division of Pediatric Infectious Diseases, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
Nicholson
Richard C
RC
Gonzalez
Gustavo V
GV
Simmons
Charles F
CF
Gombart
Adrian
A
Smith
Roger
R
Equils
Ozlem
O
D033942:Adaptor Proteins, Vesicular Transport; D000942:Antigens, Bacterial; D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000494:pharmacology; D003346:Corticotropin-Releasing Hormone / Q000378:metabolism*; D005260:Female; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008156:Luciferases; D053594:Myeloid Differentiation Factor 88 / Q000502:physiology*; D007752:Obstetric Labor, Premature / Q000503:physiopathology; D010920:Placenta / Q000502:physiology; D011247:Pregnancy; D011971:Receptors, Immunologic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051197:Toll-Like Receptor 4 / Q000502:physiology*; D014162:Transfection; D014327:Trophoblasts / Q000187:drug effects* / Q000502:physiology*
D033942:Adaptor Proteins, Vesicular Transport; D000942:Antigens, Bacterial; D018834:Chaperonin 60; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; C507779:MYD88 protein, human; D053594:Myeloid Differentiation Factor 88; D011971:Receptors, Immunologic; C470409:TICAM1 protein, human; D051197:Toll-Like Receptor 4; D003346:Corticotropin-Releasing Hormone; D008156:Luciferases
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