Tumor necrosis factor alpha stimulates amnion prostaglandin biosynthesis primarily via an action on fatty acid cyclooxygenase.
The purpose of this study was to determine how tumor necrosis factor alpha (TNF alpha) stimulates prostaglandin E2 production in human amnion. Amnion cells were isolated from term placentae and grown to confluence in culture. Incubations were conducted in quadruplicate wells for 16 hours with TNF alpha and protein synthesis inhibitors cycloheximide and actinomycin D, or arachidonic acid, acetylsalicylic acid (ASA), or staurosporine or H7 which inhibit protein kinase C activity. Prostaglandin E2 (PGE2) was measured by radioimmunoassay and cellular protein determined. The stimulatory action of TNF alpha on amnion PGE2 production was blocked by protein synthesis inhibitors, and the addition of arachidonic acid always enhanced the stimulatory properties of TNF alpha. TNF alpha consistently induced more rapid recovery from ASA treatment, and protein kinase C inhibition attenuated the stimulatory effects of TNF alpha. These results suggest that the stimulatory action of TNF alpha on amnion PGE2 production is likely at the level of induction of fatty acid cyclooxygenase activity and is partially dependent upon activation of protein kinase C.
Pollard
J K
JK
Department of Obstetrics and Gynecology, Medical Center Hospital of Vermont, Burlington.
Mitchell
M D
MD
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000470:Alkaloids / Q000494:pharmacology; D000650:Amnion / Q000378:metabolism*; D016718:Arachidonic Acid; D001241:Aspirin / Q000494:pharmacology; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D015232:Dinoprostone / Q000096:biosynthesis*; D005260:Female; D006801:Humans; D007546:Isoquinolines / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D011247:Pregnancy; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D000470:Alkaloids; D007546:Isoquinolines; D010879:Piperazines; D014409:Tumor Necrosis Factor-alpha; D003609:Dactinomycin; D016718:Arachidonic Acid; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D003513:Cycloheximide; D011451:Prostaglandin-Endoperoxide Synthases; D011493:Protein Kinase C; D019311:Staurosporine; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate; D001241:Aspirin