dinoprost and anandamide

Prostaglandins are critical for the onset and progression of labor in mammals, and are formed by the metabolism of arachidonic acid. The products of arachidonic acid, 2-arachidonoylglycerol (2-AG), and anandamide (AEA) have a similar lipid back bone but differing polar head groups, meaning that identification of these products by immunoassay can be difficult.. In the current study, we present the use of mass spectrometry as multiplex method of identifying the specific end products of arachidonic and anandamide metabolism by human derived amnion explants treated with either an infectious agent (LPS) or inflammatory mediator (IL-1β or TNF-α).. Human amnion tissue explants treated with LPS, IL-1β, or TNF-α increased production of prostaglandin E

Topics: Amnion; Arachidonic Acid; Arachidonic Acids; Cytokines; Dinoprost; Dinoprostone; Endocannabinoids; Female; Humans; Interleukin-1beta; Lipopolysaccharides; Mass Spectrometry; Polyunsaturated Alkamides; Pregnancy; Tumor Necrosis Factor-alpha

A leading hypothesis of N-acyl ethanolamine (NAE) biosynthesis, including the endogenous cannabinoid anandamide (AEA), is that it depends on hydrolysis of N-acyl-phosphatidylethanolamines (NAPE) by a NAPE-specific phospholipase D (NAPE-PLD). Thus, deletion of NAPE-PLD should attenuate NAE levels. Previous analyses of two different NAPE-PLD knockout (KO) strains produced contradictory data on the importance of NAPE-PLD to AEA biosynthesis. Here, we examine this hypothesis with a strain of NAPE-PLD KO mice whose lipidome is uncharacterized. Using HPLC/MS/MS, over 70 lipids, including the AEA metabolite, N-arachidonoyl glycine (NAGly), the endocannabinoid 2-arachidonyl glycerol (2-AG) and prostaglandins (PGE(2) and PGF(2α)), and over 60 lipoamines were analyzed in 8 brain regions of KO and wild-type (WT) mice. Lipidomics analysis of this third NAPE-PLD KO strain shows a broad range of lipids that were differentially affected by lipid species and brain region. Importantly, all 6 NAEs measured were significantly reduced, though the magnitude of the effect varied by fatty acid saturation length and brain region. 2-AG levels were only impacted in the brainstem, where levels were significantly increased in KO mice. Correspondingly, levels of arachidonic acid were significantly decreased exclusively in brainstem. NAGly levels were significantly increased in 4 brain regions and levels of PGE(2) increased in 6 of 8 brain regions in KO mice. These data indicate that deletion of NAPE-PLD has far broader effects on the lipidome than previously recognized. Therefore, behavioral characteristics of suppressing NAPE-PLD activity may be due to a myriad of effects on lipids and not simply due to reduced AEA biosynthesis.

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Brain; Brain Stem; Cerebellum; Cerebral Cortex; Chromatography, High Pressure Liquid; Corpus Striatum; Dinoprost; Dinoprostone; Endocannabinoids; Ethanolamines; Glycerides; Glycine; Hippocampus; Hypothalamus; Lipid Metabolism; Lipids; Mesencephalon; Mice, Knockout; Phosphatidylethanolamines; Phospholipase D; Polyunsaturated Alkamides; Tandem Mass Spectrometry; Thalamus

Prematurity is the leading cause of perinatal morbidity and mortality worldwide. There is a strong causal relationship between infection and preterm births. Intrauterine infection elicits an immune response involving the release of inflammatory mediators like cytokines and prostaglandins (PG) that trigger uterine contractions and parturition events. Anandamide (AEA) is an endogenous ligand for the cannabinoid receptors CB1 and CB2. Similarly to PG, endocannabinoids are implicated in different aspects of reproduction, such as maintenance of pregnancy and parturition. Little is known about the involvement of endocannabinoids on the onset of labor in an infectious milieu. Here, using a mouse model of preterm labor induced by lipopolysaccharide (LPS), we explored changes on the expression of components of endocannabinoid system (ECS). We have also determined whether AEA and CB antagonists alter PG production that induces labor. We observed an increase in uterine N-acylphosphatidylethanolamine-specific phospholipase D expression (NAPE-PLD, the enzyme that synthesizes AEA) upon LPS treatment. Activity of catabolic enzyme fatty acid amide hydrolase (FAAH) did not change significantly. In addition, we also found that LPS modulated uterine cannabinoid receptors expression by downregulating Cb2 mRNA levels and upregulating CB1 protein expression. Furthermore, LPS and AEA induced PGF2a augmentation, and this was reversed by antagonizing CB1 receptor. Collectively, our results suggest that ECS may be involved in the mechanism by which infection causes preterm birth.

Topics: Amidohydrolases; Animals; Arachidonic Acids; Cannabinoid Receptor Antagonists; Dinoprost; Disease Models, Animal; Endocannabinoids; Female; Gene Expression Regulation; Gestational Age; Lipopolysaccharides; Mice, Inbred BALB C; Obstetric Labor, Premature; Phospholipase D; Polyunsaturated Alkamides; Pregnancy; Progesterone; Receptor, Cannabinoid, CB1; Receptor, Cannabinoid, CB2; Time Factors; Uterus

Endocannabinoids are protective in animal colitis models. As endocannabinoids also form novel prostaglandin ethanolamides (prostamides) via COX-2, we investigated the effects of prostamides and other COX-2 mediators on tissue damage in an ex vivo human mucosal explant colitis model. Healthy human colonic mucosae were incubated with pro-inflammatory cytokines TNF-α and IL-1β to elicit colitis-like tissue damage. The PGF-ethanolamide analogue, bimatoprost decreased colitis scores which were reversed by a prostamide-specific antagonist AGN 211334, but not the FP receptor antagonist AL-8810. PGF-ethanolamide and PGE-ethanolamide also reduced cytokine-evoked epithelial damage. Anandamide was protective in the explant colitis model; however COX-2 inhibition did not alter its effects, associated with a lack of COX-2 induction in explant mucosal tissue. These findings support an anti-inflammatory role for prostamides and endocannabinoids in the human colon.

Topics: Adult; Amides; Arachidonic Acids; Bimatoprost; Cloprostenol; Colitis; Colon, Sigmoid; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dinoprost; Dinoprostone; Endocannabinoids; Female; Humans; Immunohistochemistry; Interleukin-1beta; Male; Middle Aged; Oxazoles; Polyunsaturated Alkamides; Receptors, Prostaglandin; Sulfonamides; Tissue Culture Techniques; Tumor Necrosis Factor-alpha; Young Adult

It was suggested that endocannabinoids are metabolized by cyclooxygenase (COX)-2 in the spinal cord of rats with kaolin/λ-carrageenan-induced knee inflammation, and that this mechanism contributes to the analgesic effects of COX-2 inhibitors in this experimental model. We report the development of a specific method for the identification of endocannabinoid COX-2 metabolites, its application to measure the levels of these compounds in tissues, and the finding of prostamide F(2α) (PMF(2α)) in mice with knee inflammation. Whereas the levels of spinal endocannabinoids were not significantly altered by kaolin/λ-carrageenan-induced knee inflammation, those of the COX-2 metabolite of AEA, PMF(2α), were strongly elevated. The formation of PMF(2α) was reduced by indomethacin (a non-selective COX inhibitor), NS-398 (a selective COX-2 inhibitor) and SC-560 (a selective COX-1 inhibitor). In healthy mice, spinal application of PMF(2α) increased the firing of nociceptive (NS) neurons, and correspondingly reduced the threshold of paw withdrawal latency (PWL). These effects were attenuated by the PMF(2α) receptor antagonist AGN211336, but not by the FP receptor antagonist AL8810. Also prostaglandin F(2α) increased NS neuron firing and reduced the threshold of PWL in healthy mice, and these effects were antagonized by AL8810, and not by AGN211336. In mice with kaolin/λ-carrageenan-induced knee inflammation, AGN211336, but not AL8810, reduced the inflammation-induced NS neuron firing and reduction of PWL. These findings suggest that inflammation-induced, and prostanoid-mediated, enhancement of dorsal horn NS neuron firing stimulates the production of spinal PMF(2α), which in turn contributes to further NS neuron firing and pain transmission by activating specific receptors.

Topics: Action Potentials; Animals; Arachidonic Acids; Cannabinoid Receptor Modulators; Chromatography, Liquid; Cyclooxygenase 1; Cyclooxygenase 2; Dinoprost; Dinoprostone; Endocannabinoids; Evoked Potentials; Hindlimb; Inflammation; Mass Spectrometry; Membrane Proteins; Mice; Nociceptors; Pain; Polyunsaturated Alkamides; Posterior Horn Cells; Rats

Anandamide, an endocannabinoid, prostaglandins derived from cyclooxygenase-2 and nitric oxide synthesized by nitric oxide synthase (NOS), are relevant mediators of embryo implantation. We adopted a pharmacological approach to investigate if anandamide modulated NOS activity in the receptive rat uterus and if prostaglandins mediated this effect. As we were interested in studying the changes that occur at the maternal side of the fetal-maternal interface, we worked with uteri obtained from pseudopregnant rats. Females were sacrificed on day 5 of pseudopregnancy, the day in which implantation would occur, and the uterus was obtained. Anandamide (2 ng/kg, i.p.) inhibited NOS activity (P<0.001) and increased the levels of prostaglandin E(2) (P<0.001) and prostaglandin F(2α) (P<0.01). These effects were mediated via cannabinoid receptor type 2, as the pre-treatment with SR144528 (10 mg/kg, i.p.), a selective cannabinoid receptor type 2 antagonist, completely reverted anandamide effect on NOS activity and prostaglandin levels. The pre-treatment with a non-selective cyclooxygenase inhibitor (indomethacin 2.5mg/kg, i.p.) or with selective cyclooxygenase-2 inhibitors (meloxicam 4 mg/kg, celecoxib 3mg/kg, i.p.) reverted anandamide inhibition on NOS, suggesting that prostaglandins are derived from cyclooxygenase-2 mediated anandamide effect. Thus, anandamide levels seemed to modulate NOS activity, fundamental for implantation, via cannabinoid receptor type 2 receptors, in the receptive uterus. This modulation depends on the production of cyclooxygenase-2 derivatives. These data establish cannabinoid receptors and cyclooxygenase enzymes as an interesting target for the treatment of implantation deficiencies.

Topics: Animals; Arachidonic Acids; Camphanes; Cannabinoid Receptor Modulators; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dinoprost; Dinoprostone; Endocannabinoids; Female; Nitric Oxide Synthase; Polyunsaturated Alkamides; Pseudopregnancy; Pyrazoles; Rats; Rats, Wistar; Receptor, Cannabinoid, CB2; Uterus

To evaluate the bimatoprost effects in the isolated human ciliary muscle and to assess how these response can be modulated by AL8810 and SR141716A.. In a myograph system (isometric force measurement), ciliary muscles were exposed cumulatively to PGF(2alpha), latanoprost, travoprost, bimatoprost, and anandamide (0.1 nM-10 microM). Experiments were also conducted in the presence of AL8810 (FP receptor antagonist; 100 nM) or SR141716A (CB(1) receptor antagonist; 10-100 nM). Contractions were expressed as the percentage of 10 microM carbachol-induced contractions.. In quiescent tissues, concentration-response curves for bimatoprost, anandamide, PGF(2alpha,) latanoprost, and travoprost were constructed. Bimatoprost showed an important contractile effect on isolated human ciliary muscle strips (E(max) = 125% +/- 0.09%); the maximal effect was higher than that obtained with carbachol. Contractions were inhibited by SR141716A (10 and 100 nM) and AL8810 (100 nM).. This study showed evidence of direct interaction of bimatoprost with the contractility of the human ciliary muscle through interaction with cannabinoid CB(1) receptor and prostanoid FP receptors.

Topics: Adult; Amides; Antihypertensive Agents; Arachidonic Acids; Bimatoprost; Cannabinoid Receptor Modulators; Ciliary Body; Cloprostenol; Dinoprost; Endocannabinoids; Humans; In Vitro Techniques; Isometric Contraction; Latanoprost; Lipids; Muscle, Smooth; Piperidines; Polyunsaturated Alkamides; Prostaglandins F, Synthetic; Pyrazoles; Receptor, Cannabinoid, CB1; Receptors, Prostaglandin; Rimonabant; Travoprost

In the present study, the abilities of cyclooxygenated derivatives of 1-arachidonoylglycerol and related compounds to prevent the metabolism of [3H]2-oleoylglycerol and [3H]anandamide by cytosolic and membrane fractions, respectively, have been investigated. For each compound, nine concentrations (range 0.2-100 microM) were tested. 1-Arachidonoylglycerol inhibited the hydrolysis of [3H]2-oleoylglycerol with a pI50 value of 5.17+/-0.04 (maximum attainable inhibition 88%). In contrast, the 1-glyceryl esters of prostaglandin D2, E2 and F2alpha were very weak inhibitors of this hydrolysis. Similarly, prostaglandin D2, prostaglandin D2 ethanolamide and prostaglandin D2 serinol amide produced <20% inhibition of [3H]2-oleoylglycerol metabolism at any concentration tested, in contrast to previous data with arachidonic acid, anandamide and arachidonoyl serinol which are all able to inhibit metabolism of this substrate under the assay conditions used here. A similar pattern was seen for all the compounds with respect to the inhibition of [3H]anandamide hydrolysis by the membrane fractions. Thus, cyclooxygenation of the arachidonoyl side chain greatly reduces the ability of 1-arachidonoylglycerol and related compounds to inhibit the hydrolysis of [3H]2-oleoylglycerol and [3H]anandamide.

Topics: Amidohydrolases; Animals; Arachidonic Acids; Brain; Cell Fractionation; Cell Membrane; Cytosol; Diglycerides; Dinoprost; Dinoprostone; Dose-Response Relationship, Drug; Endocannabinoids; Glycerides; Hydrolysis; Inhibitory Concentration 50; Monoacylglycerol Lipases; Polyunsaturated Alkamides; Prostaglandin D2; Rats; Rats, Sprague-Dawley; Tritium

Anandamide, an endogenous agonist of cannabinoid receptors, activates various signal transduction pathways. Anandamide also activates vanilloid VR(1) receptor, which was a nonselective cation channel with high Ca(2+) permeability and had sensitivity to capsaicin, a pungent principle in hot pepper. The effects of anandamide and capsaicin on arachidonic acid metabolism in neuronal cells have not been well established. We examined the effects of anandamide and capsaicin on arachidonic acid release in rat pheochromocytoma PC12 cells. Both agents stimulated [3H]arachidonic acid release in a concentration-dependent manner from the prelabeled PC12 cells even in the absence of extracellular CaCl(2). The effect of anandamide was neither mimicked by an agonist nor inhibited by an antagonist for cannabinoid receptors. The effects of anandamide and capsaicin were inhibited by phospholipase A(2) inhibitors, but not by an antagonist for vanilloid VR(1) receptor. In PC12 cells preincubated with anandamide or capsaicin, [3H]arachidonic acid release was marked and both agents were no more effective. Co-addition of anandamide or capsaicin synergistically enhanced [3H]arachidonic acid release by mastoparan in the absence of CaCl(2). Anandamide stimulated prostaglandin F(2alpha) formation. These findings suggest that anandamide and capsaicin stimulated arachidonic acid metabolism in cannabinoid receptors- and vanilloid VR(1) receptor-independent manner in PC12 cells. The possible mechanisms are also discussed.

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Calcium Chloride; Capsaicin; Dinoprost; Dose-Response Relationship, Drug; Endocannabinoids; Enzyme Activators; Extracellular Space; Intercellular Signaling Peptides and Proteins; Ionomycin; Ionophores; PC12 Cells; Peptides; Phospholipases A; Polyunsaturated Alkamides; Rats; Receptors, Cannabinoid; Receptors, Drug; Wasp Venoms