title: Stimulative effect of cadmium on prostaglandin E2 production in primary mouse osteoblastic cells.
abstract: We have reported that cadmium (Cd) stimulates bone resorption via prostaglandin E2 (PGE2), which is mainly produced in osteoblasts. Prostaglandin (PGs) is regulated by arachidonic acid (AA) release by phospholipase A2 (PLA2) and its conversion to PGs by cyclooxygenase (COX). In the present study, we investigated the possibility that Cd-induced PGE2 synthesis was mediated through PLA2 or COX or both using primary mouse osteoblastic cells in serum-free medium. Cd at 1 microM and above stimulated 14C-AA release from 14C-AA-prelabeled osteoblastic cells. PLA2 activity of cytosolic fraction in Cd-treated cells preferentially hydrolyzed AA at the Sn2 position of phospholipids and was inhibited by arachidonyltrifluoromethyl ketone (AACOCF3), an inhibitor of cytosolic PLA2 (cPLA2). Cd at 1 microM and above increased cPLA2 activity and the level of constitutive cPLA2 mRNA. Secretory PLA2 mRNA was not detected. On the other hand, Cd at 1 microM and above stimulated PGE2 production and its production was inhibited by an inhibitor of COX-2 (NS-398). Cd at 1 microM and above markedly stimulated COX-2 mRNA expression and slightly increased the level of COX-1 mRNA. An inhibitor of COX-1 (varelylsalicylic acid) did not affect Cd-induced PGE2 production. In addition, Cd-induced PGE2 synthesis was inhibited by AA-COCF3, On the other hand, IL-1 alpha, an inducer of COX-2, did not stimulated PGE2 production in present culture system. When IL-1 alpha- or Cd-treated cells were incubated with AA for 10 minutes, IL-1 alpha-treated cells as well as Cd-treated ones caused an increase in PGE2 production. This suggests that the mechanism of Cd-induced PGE2 production is different from that of IL-1 alpha, which may require an activation of cPLA2. From these results, it was found that Cd by itself stimulated PGE2 production by two successive steps that Cd increased cPLA2 activity and then COX-2 induction.
authors.affiliation: Department of Toxicology, Faculty of Pharmaceutical Sciences, Toyama Medical & Pharmaceutical University, Toyama 939-0194, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Miyahara
authors.forename: H
authors.initials: H
authors.lastname: Tonoyama
authors.forename: M
authors.initials: M
authors.lastname: Watanabe
authors.forename: A
authors.initials: A
authors.lastname: Okajima
authors.forename: S
authors.initials: S
authors.lastname: Miyajima
authors.forename: T
authors.initials: T
authors.lastname: Sakuma
authors.forename: N
authors.initials: N
authors.lastname: Nemoto
authors.forename: K
authors.initials: K
authors.lastname: Takayama
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D016718:Arachidonic Acid / Q000378:metabolism; D001095:Arachidonic Acids / Q000494:pharmacology; D002104:Cadmium / Q000494:pharmacology*; D002478:Cells, Cultured; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D015232:Dinoprostone / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins; D051379:Mice; D008815:Mice, Inbred Strains; D009578:Nitrobenzenes / Q000494:pharmacology; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010741:Phospholipases A / Q000037:antagonists & inhibitors; D054467:Phospholipases A2; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology
chemical_list: D001095:Arachidonic Acids; D004791:Enzyme Inhibitors; D007375:Interleukin-1; D007527:Isoenzymes; D008565:Membrane Proteins; D009578:Nitrobenzenes; D012333:RNA, Messenger; D013449:Sulfonamides; D002104:Cadmium; C081565:arachidonyltrifluoromethane; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; D016718:Arachidonic Acid; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; C496512:Ptgs1 protein, mouse; D010741:Phospholipases A; D054467:Phospholipases A2; D015232:Dinoprostone