Tumor necrosis factor-alpha induced the release of interleukin-6 from endometriotic stromal cells by the nuclear factor-kappaB and mitogen-activated protein kinase pathways.
OBJECTIVE
To examine the involvement of nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein kinase (MAPK) in the induction of interleukin-6 (IL-6) by tumor necrosis factor-alpha (TNF-alpha) in endometriotic stromal cells (ESC).


METHODS
Prospective study.


METHODS
Department of Obstetrics and Gynecology, Tottori University Hospital, Yonago, Japan.


METHODS
Twelve patients who underwent laparoscopic surgery.


METHODS
Endometriotic stromal cells were obtained from chocolate cyst linings of the ovary.


METHODS
We determined the effect of TNF-alpha on the production of IL-6 and the effect of inhibitors for NF-kappaB and the MAPK pathway on IL-6 production using ELISA. Western blottings and electrophoretic mobility shift assays were used to detect activation of NF-kappaB and extracellular signal-regulated kinase 1/2 (ERK1/2).


RESULTS
The addition of TNF-alpha (0.1 ng/mL) significantly increased IL-6 protein in endometriotic stromal cells. Western blottings and electrophoretic mobility shift assays revealed that incubation with TNF-alpha induced degradation of inhibitor kappaB (I kappaB) and expression of phosphorylated ERK1/2. The NF-kappaB inhibitor (TPCK) and MAPK inhibitor (U0126) blocked the TNF-alpha-induced IL-6 expression. Electrophoretic mobility shift assay revealed that U0126 attenuated activator protein-1 (AP-1) activation induced by TNF-alpha.


CONCLUSIONS
These findings demonstrate that NF-kappaB and AP-1 activation is critical for TNF-alpha-induced IL-6 expression in endometriotic stromal cells. Novel therapeutic modalities targeting these molecules may be possible in the near future.
Yamauchi
Nobuhiro
N
Department of Obstetrics and Gynecology, Tottori University School of Medicine, Yonago, Japan.
Harada
Tasuku
T
Taniguchi
Fuminori
F
Yoshida
Souichi
S
Iwabe
Tomio
T
Terakawa
Naoki
N
D015153:Blotting, Western; D002070:Butadienes / Q000494:pharmacology; D004586:Electrophoresis; D004715:Endometriosis / Q000378:metabolism* / Q000473:pathology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D015850:Interleukin-6 / Q000096:biosynthesis / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology; D010766:Phosphorylation; D011446:Prospective Studies; D017154:Stromal Cells / Q000378:metabolism* / Q000473:pathology; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D000093242:Interleukin-6 Inhibitors
D002070:Butadienes; D004791:Enzyme Inhibitors; D020672:I-kappa B Proteins; D015850:Interleukin-6; D020928:Mitogen-Activated Protein Kinases; D016328:NF-kappa B; D009570:Nitriles; D018808:Transcription Factor AP-1; D014409:Tumor Necrosis Factor-alpha; D000093242:Interleukin-6 Inhibitors; C113580:U 0126