title: Effects of anoxic stress on prostaglandin H synthase isoforms in piglet brain.
abstract: We examined effects of ischemia and asphyxia on levels of prostaglandin H synthase-1 (PGHS-1) and prostaglandin H synthase-2 (PGHS-2) in piglet brain. Ischemia was induced by increasing intracranial pressure and asphyxia was induced by turning off the respirator. Duration of anoxic stress was 10 min. In some animals, indomethacin (5 mg/kg, i.v.) or 7-nitroindazole (7-NI) was administered prior to ischemia to block PGHS or brain nitric oxide synthase (bNOS), respectively. Tissues from cerebral cortex and hippocampus were removed and fixed and/or frozen after 1, 2, 4 and 8 h of recovery from anoxic stress. In addition, tissues were obtained from untreated animals or from time control animals. Levels of mRNA and proteins were determined using RNase protection assay and immunohistochemical approaches, respectively. In the tissues studied, only a few neurons were immunopositive for PGHS-1, and neither ischemia or asphyxia affected PGHS-1 immunostaining at 8 h after recovery. Likewise, PGHS-1 mRNA did not increase following anoxic stress. In contrast, substantial PGHS-2 immunoreactivity was present in neurons and glial cells in the cerebral cortex and hippocampus and there was no difference between time control and non treated animals. PGHS-2 mRNA increased by 2-4 h after ischemia, and heightened immunoreactivity for PGHS-2 was present at 8 h after ischemia in cerebral cortex and hippocampus. However, asphyxia did not increase PGHS-2 mRNA or immunostaining. Indomethacin pretreatment inhibited increases in mRNA and protein for PGHS-2 after ischemia, while 7-NI had little effect on increases in PGHS-2 immunoreactivity. We conclude that: (1) PGHS-2 is the predominant isoform present in piglet cerebral cortex and hippocampus; (2) Ischemia but not asphyxia increases levels of PGHS-2; (3) Ischemia does not increase levels of PGHS-1; and (4) Indomethacin but not 7-NI attenuates ischemia-induced increases in PGHS-2.
authors.affiliation: Department of Opthalmology, Albert Szent-Györgyi Medical University, Szeged, Hungary.
authors.forename: R
authors.initials: R
authors.lastname: Dégì
authors.forename: F
authors.initials: F
authors.lastname: Bari
authors.forename: N
authors.initials: N
authors.lastname: Thrikawala
authors.forename: T C
authors.initials: TC
authors.lastname: Beasley
authors.forename: C
authors.initials: C
authors.lastname: Thore
authors.forename: T M
authors.initials: TM
authors.lastname: Louis
authors.forename: D W
authors.initials: DW
authors.lastname: Busija
mesh_terms: D000818:Animals; D001831:Body Temperature; D001921:Brain / Q000201:enzymology*; D002540:Cerebral Cortex / Q000187:drug effects / Q000201:enzymology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006624:Hippocampus / Q000187:drug effects / Q000201:enzymology; D002534:Hypoxia, Brain / Q000201:enzymology*; D007150:Immunohistochemistry; D007191:Indazoles / Q000494:pharmacology; D007213:Indomethacin / Q000494:pharmacology; D007527:Isoenzymes / Q000096:biosynthesis*; D008297:Male; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis*; D012260:Ribonucleases / Q000378:metabolism; D013552:Swine
chemical_list: D016861:Cyclooxygenase Inhibitors; D004791:Enzyme Inhibitors; D007191:Indazoles; D007527:Isoenzymes; D019001:Nitric Oxide Synthase; D011451:Prostaglandin-Endoperoxide Synthases; D012260:Ribonucleases; C080122:7-nitroindazole; D007213:Indomethacin