title: Effect of ceramide analogs on interleukin-1alpha-induced production of prostaglandin E2 by amnion-derived (WISH) cells.
abstract: BACKGROUND
  Prostaglandin E2 (PGE2) is reportedly synthesized in the amnion, and its levels are increased during labor. Our objective was to measure the level of PGE2 induced by interleukin (IL)-1alpha following treatment with ceramide analogs in amnion-derived cells.
  
  
  METHODS
  Amnion-derived (WISH) cells were cultured and stimulated by IL-1alpha, IL-1 receptor antagonist (ra), C2-ceramide and C6-ceramide. The levels of PGE2 in the media were measured by ELISA. The induction of prostaglandin H synthase (PGHS)-2 mRNA was detected by reverse transcription and polymerase chain reaction (RT-PCR).
  
  
  RESULTS
  Following stimulation with IL-1alpha, the production of PGE2 could not be detected until incubation had continued for 2 h, but this production appeared to continue after 4 h of incubation. The production of PGE2 was significantly increased by IL-1alpha, and was suppressed by IL-1 ra, in a dose-dependent manner. PGE2 production was significantly increased by IL-1alpha and C2-ceramide as compared with IL-1alpha alone. However, PGE2 production was not significantly increased by IL-1alpha and C6-ceramide as compared with IL-1alpha alone. PGHS-2 mRNA were induced by treatment with IL-1alpha, and were strongly induced by treatment with IL-1alpha and C2-ceramide by RT-PCR.
  
  
  CONCLUSIONS
  Results suggest that IL-1alpha induce the PGHS-2 mRNA and stimulate the production of PGE2 by a mechanism that involves the sphingomyelin-ceramide system. Ceramide may be important in increasing the production of PGE2 during parturition.
authors.affiliation: Department of Obstetrics and Gynecology, Oita Medical University, Hasama-machi, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Nakamura
authors.forename: Y
authors.initials: Y
authors.lastname: Kawano
authors.forename: S
authors.initials: S
authors.lastname: Kamihigashi
authors.forename: T
authors.initials: T
authors.lastname: Sugano
authors.forename: H
authors.initials: H
authors.lastname: Narahara
authors.forename: I
authors.initials: I
authors.lastname: Miyakawa
mesh_terms: D000650:Amnion / Q000502:physiology*; D018929:Cell Culture Techniques; D002518:Ceramides / Q000494:pharmacology*; D015232:Dinoprostone / Q000096:biosynthesis*; D005260:Female; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D007743:Labor, Obstetric; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D002518:Ceramides; D007375:Interleukin-1; D012333:RNA, Messenger; D015232:Dinoprostone