Biosynthesis Of Prostaglandin D2 Research Articles

Dexamethasone protects neonatal hypoxic-ischemic brain injury via L-PGDS-dependent PGD2-DP1-pERK signaling pathway.

Background and PurposeGlucocorticoids pretreatment confers protection against neonatal hypoxic-ischemic (HI) brain injury. However, the molecular mechanism remains poorly elucidated. We tested the hypothesis that glucocorticoids protect against HI brain injury in neonatal rat by stimulation of lipocalin-type prostaglandin D synthase (L-PGDS)-induced prostaglandin D2 (PGD2)-DP1-pERK mediated signaling pathway.MethodsDexamethasone and inhibitors were administered via intracerebroventricular (i.c.v) injections into 10-day-old rat brains. Levels of L-PGD2, D prostanoid (DP1) receptor, pERK1/2 and PGD2 were determined by Western immunoblotting and ELISA, respectively. Brain injury was evaluated 48 hours after conduction of HI in 10-day-old rat pups.ResultsDexamethasone pretreatment significantly upregulated L-PGDS expression and the biosynthesis of PGD2. Dexamethasone also selectively increased isoform pERK-44 level in the neonatal rat brains. Inhibitors of L-PGDS (SeCl4), DP1 (MK-0524) and MAPK (PD98059) abrogated dexamethasone-induced increases in pERK-44 level, respectively. Of importance, these inhibitors also blocked dexamethasone-mediated neuroprotective effects against HI brain injury in neonatal rat brains.ConclusionInteraction of glucocorticoids-GR signaling and L-PGDS-PGD2-DP1-pERK mediated pathway underlies the neuroprotective effects of dexamethasone pretreatment in neonatal HI brain injury.

Theoretical studies on model reaction pathways of prostaglandin H2 isomerization to prostaglandin D2/E2

Model reaction mechanisms in the biosynthesis of prostaglandin D2 (PGD2) and prostaglandin E2 (PGE2) from prostaglandin H2 with PGD2/E2 synthase were examined using the ab initio second-order Moller–Plesset perturbation method and density functional theory. The reaction was modeled similar to the isomerization of 2,3-dioxabicyclo[2.2.1]heptane to 3-hydroxycyclopentanone in the presence of MeS−. An explicit solvation of two H2O molecules was also considered, and two probable types of reaction mechanisms were demonstrated. One mechanism starts with proton abstraction from an oxygen-bound carbon at the endoperoxide by a thiolate ion and the other is stepwise and involves attack of a thiolate anion on an oxygen of the endoperoxide group in the first step with protonation of the other oxygen, followed by deprotonation from a carbon-attached oxygen to break an O–S bond to yield PGD2 or PGE2. We also found that the mPW1LYP hybrid method was superior to the B3LYP functional for systems with respect to the state-of-the-art CCSD(T) energetics.

Tetranor PGDM, an Abundant Urinary Metabolite Reflects Biosynthesis of Prostaglandin D2 in Mice and Humans

Prostaglandin D(2) (PGD(2)) is a cyclooxygenase (COX) product of arachidonic acid that activates D prostanoid receptors to modulate vascular, platelet, and leukocyte function in vitro. However, little is known about its enzymatic origin or its formation in vivo in cardiovascular or inflammatory disease. 11,15-dioxo-9alpha-hydroxy-2,3,4,5-tetranorprostan-1,20-dioic acid (tetranor PGDM) was identified by mass spectrometry as a metabolite of infused PGD(2) that is detectable in mouse and human urine. Using liquid chromatography-tandem mass spectrometry, tetranor PGDM was much more abundant than the PGD(2) metabolites, 11beta-PGF(2alpha) and 2,3-dinor-11beta-PGF(2alpha), in human urine and was the only endogenous metabolite detectable in mouse urine. Infusion of PGD(2) dose dependently increased urinary tetranor PGDM > 2,3-dinor-11beta-PGF(2alpha) > 11beta-PGF(2alpha) in mice. Deletion of either lipocalin-type or hemopoietic PGD synthase enzymes decreased urinary tetranor PGDM. Deletion or knockdown of COX-1, but not deletion of COX-2, decreased urinary tetranor PGDM in mice. Correspondingly, both PGDM and 2,3-dinor-11beta-PGF(2alpha) were suppressed by inhibition of COX-1 and COX-2, but not by selective inhibition of COX-2 in humans. PGD(2) has been implicated in both the development and resolution of inflammation. Administration of bacterial lipopolysaccharide coordinately elevated tetranor PGDM and 2,3-dinor-11beta-PGF(2alpha) in volunteers, coincident with a pyrexial and systemic inflammatory response, but both metabolites fell during the resolution phase. Niacin increased tetranor PGDM and 2,3-dinor-11beta-PGF(2alpha) in humans coincident with facial flushing. Tetranor PGDM is an abundant metabolite in urine that reflects modulated biosynthesis of PGD(2) in humans and mice.

Biosynthesis of prostaglandin D2 by motoneurons and dorsal horn microneurons: a biochemical and high resolution immunocytochemical study in chick spinal cord.

Prostaglandin D2 is one of the major prostanoids formed from [14C]arachidonic acid by the central nervous system. The aim of the present study is to specify the prostaglandin D2 biosynthetic capacity in the chick spinal cord and to identify the cell type involved in this synthesis. A highly specific and sensitive enzyme immunoassay allowed us to demonstrate that the amount of newly formed prostaglandin D2 increases proportionally with the concentration of free arachidonic acid of either exogenous or endogenous origin and reaches concentration values ranging from 10(-9) to 10(-6) M. The sites of prostaglandin D2 synthesis were localized in Vibratome sections of spinal cord after incubation with antibodies raised against glutathione-independent prostaglandin D synthase; controls were performed with anti-glutathione-dependent prostaglandin D synthase antibodies and non-immune rabbit or goat serum. After immunoprocessing, electron microscope examination revealed that the specific immunoreactivity was confined to small neurons of laminae II and III in the dorsal horn and to motoneurons in the ventral horn of the spinal cord. The immunodeposits were associated with rough endoplasmic reticulum profiles distributed throughout the dorsal horn neurons or restricted to limited subsurface areas of perikarya and dendrites in motoneurons. Since the immunoreactive neurons in the dorsal horn were closely related to blood capillaries, prostaglandin D2 may be suspected to play a role in the regulation of the microcirculation. The accumulation of prostaglandin D synthase in motoneuron areas facing astrocytic membrane stacks suggests that prostaglandin D2 could interact with astrocytic functions.